CN117999282A - Anti-TCR delta variable 1 antibodies - Google Patents

Abstract

The present invention provides high affinity anti-TCR delta variable 1 (anti-vδ1) antibodies and antibody fragments thereof. The invention also provides compositions and pharmaceutical compositions comprising such antibodies, and methods of making such antibodies. The invention also provides therapeutic methods and medical uses involving the antibodies.

Description

Anti-TCR delta variable 1 antibodies

Technical Field

The present invention relates to antibodies and fragments and variants thereof that specifically bind to T cell receptors of γδ T cells.

Background

The growing interest in T cell immunotherapy of cancer has focused on the apparent ability of subsets of cd8+ and cd4+ alpha beta (alpha beta) T cells to recognize cancer cells and mediate the potential for host protective functions, particularly when the inhibitory pathways of PD-1, CTLA-4 and other receptors are inhibited by clinically mediated antagonism. However, αβ T cells are MHC restricted, which may lead to graft versus host disease.

Γδ T cells (GAMMA DELTA T cells) represent a subset of T cells expressing a unique, defined γδ T Cell Receptor (TCR) on the surface. The TCR consists of one gamma (gamma) chain and one delta (delta) chain, each of which undergoes chain rearrangement but has a limited number of V genes compared to αβt cells. The major TRGV gene segments encoding vγ are TRGV2, TRGV3, TRGV, TRGV5, TRGV8, TRGV9 and TRGV11, and nonfunctional genes TRGV10, TRGV11, TRGVA and TRGVB. The most common TRDV gene segments encode vδ1, vδ2, and vδ3, plus several V segments with both vδ and vα names (Adams et al 296:30-40 (2015) Cell immunol.). Human γδ T cells can be broadly classified according to their TCR chains, as certain γ and δ types are more commonly (but not exclusively) found on cells in one or more tissue types. For example, most blood-resident γδ T cells express vδ2 TCR, typically vγ9vδ2, which is less common in tissue-resident γδ T cells (such as those in the skin), which more frequently use vδ1 TCR paired with the γ chain (e.g. often paired with vγ4 in the gut).

Γδ T cells play a key role in immune monitoring, recognizing malignant or transformed cells (e.g., cancer cells) through stress marker patterns, and then exert potent and selective cytotoxicity. Thus, γδ T cells can act as a coordinator of immune responses. Even in tumors with low mutational burden, which have proven challenging for other immunotherapies, in situ modulation of these cells offers the potential to increase immunogenicity. The recognition of tumors by γδ T cells is not dependent on any single tumor antigen, and therefore modulators of γδ T cells have potential in a range of disease indications, including both hematological and solid malignancies. The recognition mechanism of γδ T cells is not MHC restricted.

The authors of WO2019147735 hypothesize that some γδ cells have a pro-tumor activity or inhibit αβ T cell mediated anti-cancer immune responses. The authors postulate that γδ T cells are immunosuppressants and therefore suggest that they should be depleted, inhibited or blocked by antibodies in the cancer setting.

However, while anti- γδ antibodies are generally believed to negatively regulate γδ cell function by blocking or killing such cells, positive correlation between γδ T cell infiltration and prognosis and/or survival of patients has been found.

In contrast to the αβ TCR receptor/ligand interaction, the knowledge of the vδ TCR receptor/ligand interaction is limited. In the absence of such insight, antibodies recognizing vδ TCR to date have been the primary exploratory tool for such interactions. Such tools are typically crude blocking antibodies that demonstrate that TCR receptor/ligand interactions result in blocking, inhibition, or ablation of vδ1+ cells. For example, tool antibodies TS8.2 and TS-1 were used in the study as anti- γδ blocking antibodies, indicating that the antibodies reduced cytotoxicity of vδ1 cells. These studies, in combination with others, demonstrate that the use of such anti-vδ1 antibodies to advantageously modulate the cytotoxicity of vδ1 cells in an in situ disease environment is not envisioned, and thus antibodies that increase rather than decrease the cytotoxicity of vδ1 are desirable.

For immunotherapy with γδ T cells, a means is needed to expand the cells in situ, or to harvest them and expand them ex vivo prior to reinfusion. The latter method has been previously described, which uses the addition of exogenous cytokines, see for example WO2017/072367 and WO2018/212808. Methods of expanding patient autologous γδ T cells have been described that employ a pharmaceutically modified form of hydroxy-methylbut-2-enyl pyrophosphate (HMBPP) or a clinically approved aminobisphosphonate. By these methods, more than 250 cancer patients were treated, seemingly safe, but with little chance of complete remission. However, there remains a need for activators with a confirmed ability to expand large numbers of γδ T cells.

Furthermore, binding agents or activators that are capable of preferentially targeting or binding or recognizing or specifically modulating or increasing the number of vδ1+ cells in situ may be highly desirable as drugs.

However, while there are drugs potentially modulating vδ2+ cells, including aminobisphosphonates such as(Zoledronic acid), but the drug is designed primarily to slow bone resorption. Regardless of the vδ2+ modulation, it is desirable to develop drugs specifically designed to bind, target, modulate, activate or increase vδ1+ cell numbers. This is because, for example, repeated vδ2+ modulation may result in a durable and gradually depleting phenotype.

Furthermore, given the tissue-resident nature of vδ1+ cells, ideal drugs that are able to modulate vδ1+ will also exhibit fewer 'off-target' adverse reactions and rapid renal clearance. In general, the adverse reactions may occur when small molecule chemicals are employed. For example, the foregoing amino bisphosphonates, which are shown to be capable of modulating a separate class of V.delta.2+ cells as a secondary to the primary modulating effect on bone, are associated with nephrotoxicity manifested as worsening renal function and potential renal failure (e.g., markowitz et al (2003) Kidney int.64 (1): 281-289). Other adverse effects of Zometa listed by the European drug administration include anemia, anaphylaxis, hypertension, atrial fibrillation, myalgia, general pain, discomfort, increased blood urea, vomiting, joint swelling, chest pain, and the like.

The in situ environment in which vδ1+ cells themselves are located must be further considered. For example, it has been previously shown that non-hematopoietic tissue resident γδ T cells show a strong proliferative response when first isolated from tissue, but provided they are not in direct contact with autologous fibroblasts. It has been found that non-hematopoietic tissue resident T cells (γδ T cells) must be isolated from non-hematopoietic cells (e.g., stromal cells, particularly fibroblasts) to function. This is because direct contact of lymphocytes with stromal cells or epithelial cells appears to inhibit the expansion of tissue resident γδ T cells. The observation that pre-activated cells exist in situ in a further inhibited state is another reason that vδ1 cells have not been considered as promising therapeutic targets to date. Indeed, until the findings described herein, it was not envisaged how these cells could be advantageously and selectively regulated in situ, with blood and tissue vδ1+ cells generally considered as 'quiescent', 'pre-activated' or 'non-activated'.

Bispecific and multispecific antibodies in various formats have been developed for various therapeutic uses. Bispecific and multispecific antibodies can be classified into separate (albeit overlapping) classes based on the type and mode of action of the biological target. For example, such multispecific antibodies can be divided into classes such as cytotoxic effector cell redirecting agents (also known as bispecific T cell recruiting antibodies, bispecific T cell adaptors, TCEs, or bites) and Dual Immunomodulators (DI).

TCE (T cell adapter) is intended to enhance the immune response of a patient to a tumor by targeting T cells to tumor cells or vice versa, and to act by targeting a first epitope of the T cell receptor complex of T cells (typically CD 3) and a second epitope that is a cancer antigen or a cancer related antigen such as a tumor related antigen (TAA). Such antibodies co-localize tumor cells and T cells to promote tumor cell killing. Examples of bites include the CD3 x CD19 bispecific antibody bolafumab (blinatumomab), the CD3 x EpCAM bispecific antibody katuxomab (catumaxomab), and the CD3 x HER2 bispecific antibody ertuxomab (ertumaxomab). TCEs such as BiTE are typically provided in scFv formats, but other formats have been provided. For example, bikes are similar to BiTE, but they target CD16 on NK cells, rather than CD3.

T cell receptors have been described as the most complex receptor structure in the mammalian immune system. It constitutes a transmembrane polyprotein receptor complex that contains T-cell receptors in close proximity to many CD3 chains. For example, in mammals, a typical such complex comprises one T cell receptor, one CD3 gamma chain, one CD3 delta chain, and two CD3g chains. These chains associate with the T Cell Receptor (TCR) and zeta chain (zeta chain), combine, and then generate typical activation signals in T lymphocytes. However, alternative complexes have also been reported. For example, T cell receptor complexes comprising T cell receptors and zeta chain homodimers have been described. Other co-receptors such as CD4 and CD8 may also assist TCR function.

Regardless of the composition of the receptor complex, it has been well established that the complex converts cell surface binding events to intracellular phosphorylation signalling cascades. These phosphorylation events ultimately lead to activation of transcription factors (e.g., NFAT and NFkB), resulting in increased expression of cytokines and effector proteins (e.g., granzymes and perforins).

However, while the use of such TCEs for the treatment of cancer remains an attractive concept, to date, many of these bispecific antibodies exhibit poorly curable safety, efficacy and manufacturability profiles even after 30 years of joint effort to advance TCEs in early clinical development. In fact, by month 1 of 2020, bordetention remains the only undirected TCE in bulk. This TCE multispecific antibody fragment binds to the T cell receptor complex on a first binding arm and to the CD19 target on a second binding arm.

Bispecific T cell recruiting antibodies are discussed in Lejeune et al, 202O,Front Immunol, 11: 762. However, given the efficacy of CD3 antigen as a signal transducer and its ubiquity in patient T cell populations, existing bispecific antibodies in this class, particularly those that recruit T cells via CD3 binding, have significant off-target effects, leading to serious adverse effects. Thus, for bispecific examples targeting CD3, such as cetuximab (now withdrawn), systemic delivery (e.g., intravenous) is not practical. Instead, more inclusive delivery is more often considered, such as intraoperative, intraperitoneal, and the like. This in turn limits the selectivity and usefulness of the bispecific antibody as a medicament. Indeed, even effector-attenuated anti-CD 3 antibodies (i.e., CD 3-targeting T cell complex adaptors but not bispecific), the associated toxicity makes intravenous delivery challenging. For example, to limit exposure and reduce toxicity, the anti-CD 3 antibody, fu Lei Lushan antibody (Foralumab), is now most often considered for oral delivery (e.g., in the treatment of intestinal diseases).

It is often stated that many of the frustration observed in early clinical trials using such TCEs is currently due to the high affinity T cell complex binding domains employed. Furthermore, it is suggested that this is because those designing these TCEs do not properly consider the low affinity of the native TCR complex binding events, which are hindered by severe dose limiting toxicity, resulting in a treatment window that is too narrow. In connection with this, it is emphasized that many early TCE drug developers relied on three anti-CD 3T cell complex binding domains derived from OKT3, SP34 and UCHT 1. And these original binding domains all bind with a relatively high affinity in the one-digit to two-digit nM range, which corresponds to an affinity approximately 1,000 times higher than the native binding event. In turn, it has been proposed that this can result in a distinct (and often detrimental) effect on T cell activation compared to the natural binding of the T cell receptor complex. For example, TCE developers using a higher affinity OKT 3-based platform may be confused by the fact that OKT3 has an apoptotic effect on T cells in the presence of IL-2.

For these reasons, it is clear that lower affinity T cell complex binding is an important consideration in determining T cell engagement bispecific antibody therapeutic design parameters.

Another problem in designing the TCE is the need to attenuate Fc function. In fact, TCE is often required to fully inhibit Fc-mediated effector function to maximize therapeutic efficacy and minimize off-target toxicity, as Fc binding to fcγ receptor (fcγr) results in activation of immune effector cells. Indeed, most CD 3-targeting bispecific antibodies in current clinical practice have an Fc domain with reduced binding activity to fcγr, or are bispecific fragments that are intentionally devoid of an Fc region. TCEs with unabated Fc function are generally expected to induce antibody dependent cell-mediated cytotoxicity (ADCC) to deplete antibody-recognized γδ T cell populations. However, again, by attenuating this function to avoid toxicity/safety complications, it is also possible to attenuate the potentially important efficacy perspective, for example by engaging cd16+ or cd32+ or cd64+ immune cells, or by reducing the half-life of the bispecific antibody (e.g., if smaller bispecific antibody fragments, such as BITE, are used). Methods of reducing fcγr and TCE interactions (e.g., using IgG formats intended to reduce the interactions) are expected to reduce Fc-mediated TCE immobilization and reduce TCR aggregation by cross-linking with the immobilized TCE.

To address some but not all of these complications, many companies such as Xencor (Pasadena, CA), macrogenics (Gaithersburg, MD) and Genentech (San Francisco, CA) have recently reported decreasing the binding affinity of T cell receptor complex binding arms in their respective TCE platforms. However, decreasing the affinity of this binding can lead to reduced effectiveness and reduced selectivity in TCE design and functionality. For example, affinity-driven in vivo distribution characteristics of binding domains in such TCEs are now demonstrated. In particular, TCE distribution is often observed biased towards its highest affinity target. Thus, by decreasing the affinity of the TCE binding domain to the T cell complex, a distribution away from T cells is generally favored; these T cells are the cells required to drive the efficacy of such TCEs. Partly for these reasons, the TCE therapeutic window is referred to as 'too narrow'.

Furthermore, there are significant additional barriers to immunomodulatory therapeutic agents, particularly for solid tumors. For example, the current state of the art approach includes bispecific antibodies targeting CD3, wherein a first domain binds CD3 and a second domain targets TAA. However, these often prove problematic. For example, middelburg J et al Overcoming Challenges for CD-Bispecific Antibody THERAPY IN Solid Tumors. Cancer.2021; 13 (2): 287 summarizes some of the disorders of such multispecific, T cell-linked immunomodulatory portions in the field of solid tumors, including:

i. Mid-target, extra-tumor toxicity problems-e.g., authors stress that solid tumor TAAs are also often expressed on tissues of healthy organs, which in turn can lead to immune pathology and organ failure, and can be fatal, as shown by preclinical mouse studies using CD3 bispecific antibodies.

Availability of effector cells in Tumor Microenvironment (TME) -for example, traditional cd3+ 'naive' T cells are predominantly present in the blood and lymphatic system, without any additional dendritic cell-mediated activation.

Considering this, the mass of tumor-infiltrating lymphocytes (TILs) may include cd3+ tregs and/or depleted cd3+ cells-e.g., bispecific cells targeting CD3 may induce TIL apoptosis by activating induced cell death, thereby impeding a strong anti-tumor response.

Thus, there is a need for improved multi-specific immunomodulatory drugs in which at least one binding domain binds T cells and at least one second binding domain targets TAAs.

There is also a need to design improved drugs for specifically targeting vδ1+ cells and for treating infections, autoimmune disorders and cancers. In particular, there is a need for agents that can be administered to improve the signs and symptoms of disease by specifically binding to vδ1+ cells, targeting vδ1+ cells, specifically activating vδ1+ cells, specifically enhancing proliferation and/or cytotoxic activity of vδ1+ cells, or specifically blocking activation of vδ1+ cells.

Disclosure of Invention

The present invention provides high affinity anti-TCR delta variable 1 (anti-vδ1) antibodies and antibody fragments thereof. The antibodies of the invention have advantageous functional characteristics. In particular, unlike prior art anti-vδ1 antibodies that focus on vδ1T cell depletion, the antibodies of the present invention may be used to activate vδ1T cells. Although they cause down-regulation of the TCR on the T cells to which they bind, they do not lead to depletion of vδ1T cells, but rather they stimulate T cells and are therefore useful in therapeutic settings that would benefit from activation of this T cell compartment. The activation of vδ1T cells is evident by TCR down-regulation, activation markers such as CD25 and Ki67, degranulation markers CD107a, NCR (natural cytotoxic receptor) and/or changes in 4-1 BB. Activation of vδ1T cells in turn leads to release of inflammatory cytokines such as infγ and tnfα to promote immune licensing. Surprisingly, antibodies with suitably high affinity for TRDV1 cause increased killing of vδ T cells, and unlike antibodies that target CD3, for example, can provide high affinity antibodies without the adverse effects associated with large scale activation by CD 3. In turn, high affinity antibodies can induce strong immunostimulation by Tumor Infiltrating Lymphocytes (TIL). This can be achieved by minimal depletion or killing of vδ1 cells. Thus, the antibodies of the invention may be considered agonistic antibodies.

According to a first aspect of the present invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

A VHCDR3, said VHCDR3 comprising a sequence identical to SEQ ID NO:55 to 78, an amino acid sequence having at least 80% sequence identity; and/or

A VLCDR3, said VLCDR3 comprising an amino acid sequence corresponding to SEQ ID NO:82 to 105, and an amino acid sequence having at least 80% sequence identity to any one of seq id nos

Or comprises:

a VHCDR3, said VHCDR3 comprising a sequence identical to SEQ ID NO:133 to 143, having at least 80% sequence identity; and/or

A VLCDR3, said VLCDR3 comprising an amino acid sequence corresponding to SEQ ID NO:147 to 157 has an amino acid sequence that has at least 80% sequence identity.

According to a second aspect of the present invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

An HCDR1 sequence comprising the sequence of GDSVSSKSX ₁ A (SEQ ID NO: 158);

An HCDR2 sequence comprising SEQ ID NO: 53;

An HCDR3 sequence comprising the sequence of X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO: 162), wherein the HCDR3 sequence is not SEQ ID NO:54;

An LCDR1 sequence comprising SEQ ID NO: 79;

an LCDR2 sequence comprising SEQ ID NO: 80; and

An LCDR3 sequence comprising a sequence of QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO: 166), wherein the LCDR3 sequence is not SEQ ID NO:81;

Wherein each of X ₁ to X ₁₃ is a naturally occurring amino acid;

Or (b)

An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

An HCDR1 sequence comprising SEQ ID NO:130 sequence of 130

An HCDR2 sequence comprising SEQ ID NO:131 sequence

An HCDR3 sequence comprising the sequence of X ₁X₂YX₃X₄ AFDI (SEQ ID NO: 183), wherein the HCDR3 sequence is not SEQ ID NO:132, a part of the material;

an LCDR1 sequence comprising SEQ ID NO:144, a sequence of seq id no;

An LCDR2 sequence comprising SEQ ID NO: 145; and

An LCDR3 sequence comprising a sequence of QQX ₅X₆X₇X₈LX₉ T (SEQ ID NO: 186), wherein the LCDR3 sequence is not SEQ ID NO:146;

wherein each of X ₁ to X ₉ is a naturally occurring amino acid.

In a third aspect of the invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, or wherein the parent anti-vδ1 antibody or antigen binding fragment thereof comprises a VL sequence comprising the amino acid sequence of SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, a VL sequence of the amino acid sequence of seq id no. Optionally, in any affinity matured variant, the first residue of the VH sequence may be Q or E. Optionally, in any affinity matured variant, the first two residues of the VL sequence may be absent in any affinity matured variant as compared to the parent sequence.

In a fourth aspect of the invention, there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises:

a) Comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

b) Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

c) Comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

d) Comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

e) Comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

f) Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

g) Comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

h) Comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

i) Comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

J) Comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, and a VL of the amino acid sequence of 313.

In a fifth aspect of the invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof comprising a kappa light chain variable sequence, wherein the residue at position 74 of the kappa light chain variable sequence is not serine according to the IMGT numbering system. In some embodiments, serine at position 74 may be substituted with a non-human germline amino acid. In some embodiments, the substitution may be a non-conservative mutation, e.g., a serine substitution to a non-polar amino acid. In some embodiments, serine can be substituted with leucine.

In a sixth aspect of the invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof which specifically binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) and competes with an antibody or antigen-binding fragment thereof of any one of the first to fifth aspects of the invention for binding to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR).

In a seventh aspect of the invention, there is provided a polynucleotide sequence encoding an anti-vδ1 antibody or antigen binding fragment thereof of the present invention. For example, a polynucleotide sequence encoding a polypeptide comprising a sequence corresponding to SEQ ID NO:199 to 222, 224 to 247, 249 to 259, or 261 to 271, or an antibody binding fragment thereof.

In an eighth aspect of the invention, there is provided an expression vector comprising a polynucleotide sequence of the invention. Also provided is a host cell comprising a polynucleotide sequence of the invention or an expression vector of the invention. Also provided is a method for producing any of the antibodies or antigen-binding fragments thereof of the invention, comprising culturing a host cell of the invention in a cell culture medium.

In another aspect of the invention, there is provided a composition comprising an antibody or antibody binding fragment thereof of the invention. Also provided is a pharmaceutical composition comprising an antibody or antibody-binding fragment thereof of the invention and a pharmaceutically acceptable diluent or carrier. The compositions and pharmaceutical compositions may also optionally comprise one or more other therapeutically active agents.

In another aspect of the invention, a kit is provided comprising an anti-vδ1 antibody of the present invention or an antibody binding fragment thereof or a pharmaceutical composition of the present invention, optionally comprising instructions for use and/or other therapeutically active agents.

In another aspect of the invention, there is provided a method of treating a disease or disorder in a subject, the method comprising administering to the subject an anti-vδ1 antibody or antibody binding fragment thereof of the present invention, or a pharmaceutical composition of the present invention. Also provided is a method of modulating an immune response in a subject, the method comprising administering to the subject an anti-vδ1 antibody or antibody binding fragment thereof of the present invention, or a pharmaceutical composition of the present invention. Administration of the antibody to the subject may be administration of a therapeutically effective amount.

In yet another aspect of the invention, a method of mutating an antibody or antigen binding fragment thereof is provided, the method comprising providing an antibody comprising a kappa light chain having serine at position 74 according to the IMGT numbering system, and replacing (e.g., mutating) serine to a different amino acid. In some embodiments, serine at position 74 may be substituted with a non-human germline amino acid. In some embodiments, the substitution may be a non-conservative mutation, e.g., a serine substitution to a non-polar amino acid. In some embodiments, serine can be substituted with leucine.

In another aspect of the invention, there is provided a method of making a variant anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising providing a parent antibody comprising:

a) Comprising SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, VL of the amino acid sequence of seq id no;

b) Comprising SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, VL of the amino acid sequence of seq id no;

c) Comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

d) Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

e) Comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

f) Comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

g) Comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

h) Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

i) Comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

j) Comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

k) Comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

L) a polypeptide comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, VL of the amino acid sequence of seq id no;

And affinity maturation of the antibody, wherein the produced antibody has an affinity for the variable 61 (vδ1) chain of γδ T Cell Receptor (TCR) that is greater than that of the parent antibody

In another aspect of the invention, there is provided a method of preparing a pharmaceutical composition comprising providing an antibody prepared according to the method of preparing a variant anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment of the invention and co-formulating the antibody with at least one or more pharmaceutically acceptable diluents or carriers.

In a further aspect of the invention there is provided an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, of the invention, or a pharmaceutical composition of the invention, or a kit of the invention, for use in medicine. Also provided is the use of an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, of the invention for the manufacture of a medicament.

Drawings

Fig. 1: ELISA detection of antigens directly coated with anti-V.delta.1 Ab (REA 173, miltenyi Biotec). Detection was performed only on those antigens containing the vδ1 domain. Leucine Zipper (LZ) format appears to be more efficient than Fc format, consistent with cell-based flow competition assays (data not shown).

Fig. 2: polyclonal phage DELFIA data for DV1 selection. (a) heterodimer selection: the heterodimeric LZ TCR format in rounds 1 and 2 was deselected in both rounds. (B) homodimer selection: round 1 was performed using homodimer Fc fusion TCRs, deselecting human IgG1 Fc, then round 2 was performed on heterodimer LZ TCRs, deselecting heterodimer LZ TCRs. Each figure contains two bars per target to represent selections from different libraries.

Fig. 3: igG capture: left) sensorgram of anti-L1 IgG interaction with L1, right) steady state fit (if available). All experiments were performed on a MASS-2 instrument at room temperature. Steady state fitting was performed according to Langmuir (Langmuir) 1:1 combinations.

Fig. 4: TCR down-regulation assay (with THP-1 preloaded antibodies) results for clones 1245_p01_e07, 1252_p01_c08, 1245_p02_g04, 1245_p01_b07 and 1251_p02_c05 (a) or clones 1139_p01_e04, 1245_p02_f07, 1245_p01_g061245_p01_g09, 1138_p01_b09, 1251_p02_g10 and 1252_p01_c08 (B).

Fig. 5: t cell degranulation assay results for clones 1245_p01_e07, 1252_p01_c08, 1245_p02_g04, 1245_p01_b07 and 1251_p02_c05 (a) or clones 1139_p01_e04, 1245_p02_f07, 1245_p01_g06, 1245_p01_g09, 1138_p01_b09 and 1251_p02_g10 (B).

Fig. 6: the results of killing assays (THP-1 flow-based assays) for clones 1245_p01_e07, 1252_p01_c08, 1245_p02_g04, 1245_p01_b07 and 1251_p02_c05 (a) or clones 1139_p01_e04, 1245_p02_f07, 1245_p01_g06, 1245_p01_g09, 1138_p01_b09 and 1251_p02_g10 (B).

Fig. 7: total cell count during experiment 1 of example 10. Samples were incubated with different concentrations of anti-vδ1 antibodies described herein and compared to samples incubated with a comparison antibody or control. The graph shows the total cell count on days (a), 7, (B), 14 and (C) 18.

Fig. 8: analysis of vδ1T cells during experiment 1 of example 10. The graph shows the percentage of (a) vδ1T cells, (B) vδ1T cell counts, and (C) fold change in vδ1 in the 18 th day samples.

Fig. 9: total cell count during experiment 2 of example 10. Samples were incubated with different concentrations of anti-vδ1 antibodies described herein and compared to samples incubated with a comparison antibody or control. The graph shows the total cell counts on (A) day 7, (B) day 11, (C) day 14, and (D) day 17.

Fig. 10: analysis of vδ1T cells during experiment 2 of example 10. The graph shows the percentage of (a) vδ1T cells, (B) vδ1T cell counts, and (C) fold change in vδ1 in the sample on day 17.

Fig. 11: cell composition analysis. Cell types (including non-vδ1 cells) present in the samples were measured on day 17 of experiment 2. Cells were harvested and analyzed for surface expression of vδ1, vδ2, and αβtcr by flow cytometry. The percentage values are also provided in table 12.

Fig. 12: SYTOX-flow kill assay results. Cell function was tested using a SYTOX-flow-killing assay and presented the results of (A) cells with an effector to target (E: T) ratio of 10:1 on day 14 of experiment 1 and (B) cells with E: T ratios of 1:1 and 10:1 on day 17 of experiment 2 (after freeze thawing).

Fig. 13: total cell count after freeze thawing. The figure shows the total cell count of cultures contacted with B07, C08, E07, G04 or OKT-3 antibodies before freezing after 7 days of cell culture after freeze thawing.

Fig. 14: cell expansion was monitored. For cells cultured after freeze thawing, total cell count was monitored until day 42.

Fig. 15: binding equivalence study of modified anti-vδ1 antibodies.

Fig. 16: binding equivalency of anti-vδ1 antibodies to human germline vδ1 antigen and polymorphic variants thereof was studied.

Fig. 17: anti-vδ1 antibodies increase cytokine secretion levels by vδ1+ cells. Tissue-derived γδ T cells were incubated with the indicated antibodies. (A) An observed level of TNF- α, (B) an observed level of IFN- γ.

Fig. 18: anti-vδ1 antibodies increase granzyme B levels/activity of vδ1+ cells. Cancer cells were co-cultured with tissue-derived γδ T cells at a set t:e ratio of 1:20 for one hour and co-cultured with the indicated antibodies. The results highlight the amount of granzyme B detected in cancer cells at the end of co-culture.

Fig. 19: anti-vδ1 antibodies confer modulation and proliferation of immune cells in human tissues. Human skin punch biopsies (from five different donors) were incubated in culture for 21 days with the indicated antibodies. (A) number of viable pan [ gamma ] [ delta ] + cells. (B) number of viable V.delta.1+ cells. (C) percentage of viable biscationic V.delta.1+CD25+ cells.

Fig. 20: anti-vδ1 antibodies confer modulation and proliferation of Tumor Infiltrating Lymphocytes (TILs) in human tumors. Investigation of Renal Cell Carcinoma (RCC) +/-antibodies: (A) fold increase in TILvδ1+ cells. (B) total number of TILvδ1+ cells. (C) example gating strategy. (D) comparison of cell surface phenotype profiles of TILvδ1+ cells. (E) analysis of TIL V.delta.1 negative gating scores.

Fig. 21: anti-vδ1 antibodies enhance vδ1+ mediated cytotoxicity and diseased cell-specific cytotoxicity. Cytotoxicity/potency assays in model systems comprising ternary cultures of vδ1+ effector cells, THP-1 monocyte cancer cells, and non-diseased, healthy primary monocytes. (A) Quantification of THP-1 and monocyte numbers in ternary co-culture with γδ T cells in the presence of anti-vδ1 mAb or control. (B) Bar graph shows, highlighting the window between diseased cell specific killing and non-diseased healthy cells: left bar graph; fold increase in killing of diseased cells (THP-1) versus non-diseased cells (primary human monocytes); right side bar graph; the same data, but expressed as percent of enhanced killing compared to control. (C) The results of the list summarize the percentage increase in efficacy of vδ1+ effector cells to kill THP-1 target cells +/-mAb. (D) Results from the list of EC50 values calculated according to panel (a) are expressed as the number of γδ T cells required to confer 50% thp-1 cell killing.

Fig. 22: multispecific antibodies enhance vδ1+ effector cell mediated cytotoxicity. Targeting of tissue-centric disease-associated antigens: (a-D) vδ1+ effector cells were co-cultured with an example of a-431 cancer cells +/-multispecific antibodies comprising an anti-vδ1×anti-TAA (EGFR) bispecific binding moiety, wherein the anti-vδ1vl+vh binding domain (for the first target) was combined with the CH1-CH2-CH3 domain of the anti-EGFR binding moiety (for the second target). (E-H) vδ1+ effector cells were co-cultured with an example of a-431 cancer cells +/-multispecific antibodies comprising an anti-vδ1×anti-TAA (EGFR) bispecific binding moiety, wherein the anti-vδ1 binding domain (for the first target) comprises a full length antibody (VH-CH 1-CH2-CH 3/VL-CL) followed by combination with an anti-EGFR cetuximab (cetuximab) -derived scFv binding moiety (for the second target). (I-J) represents an alternative method of data: the multispecific antibodies provide an increased percentage of cytotoxicity of vδ1+ effector cells to egfr+ cells relative to the constituent parts.

Fig. 23: the multispecific antibodies enhance vδ1+ mediated cytotoxicity and diseased cell-specific cytotoxicity. Targeting of hematopoietic disease-associated antigens (A) E:T ratio required to induce 50% Raji cell killing, (B) percent improvement after addition of V.delta.1-CD 19 multispecific antibody

Fig. 24: phage selection rounds. (A) round 1 to round 3 phage selection of ADT1-7 library. (B) round 1 to round 3 phage selection of ADT1-4 library. (C) Round 1 to round 3 phage selection of ADT1-4 libraries using a selection strategy to isolate cynomolgus cross-reactive binders.

Fig. 25: schematic representation of selection of mature antibodies by mammalian display. (A) schematic flow-sorting of ADT1-7 library. (B) schematic flow-sorting of ADT1-4 library 1 (human). (C) Flow-sorting schematic of ADT1-4 library 2 (cynomolgus monkey).

Fig. 26: influence of kappa chain S74L changes on affinity. (a) ADT1-4 lineage mammalian display output: human dissociation rates (SPR) bind to human antigen (DELFIA ELISA,0.4nM human antigen) and indicate the use of LC 74S (open circles) or 74L (filled circles). The results highlighted a good agreement between the increased affinity of SPR and antigen binding of DELFIA ELISA. These results also underscore the improvement in binding conferred by the variable domain S74L modification. (B) ADT1-4 lineage mammalian display output: cynomolgus monkey dissociation rate (SPR) binds to cynomolgus monkey antigen (DELFIA ELISA,10nM cynomolgus monkey antigen) and indicates the use of LC 74S (open circles) or 74L (filled circles). (C) ADT1-4 lineage mammalian display output: the human antigen dissociation rate (SPR) increased the correlation with cynomolgus monkey antigen dissociation rate (SPR) plus LC 74S (open circles) or 74L (filled circles) use. (D) The effect of S74 and L74 on ADT1-4 lineage affinities was further confirmed. Mutating the starting serine at kappa chain position 74 in the lower affinity parent ADT1-4 to leucine to produce the kappa chain SEQ ID NO:504. the modified light chain was paired with the starting ADT1-4VH (SEQ ID NO: 1) to create a new molecule "ADT1-4 (S74L)", as shown. Reversion of the starting leucine at kappa chain position 74 of high affinity ADT1-4-2 to serine to produce kappa chain SEQ ID NO:505. it was paired with ADT1-4-2VH (SEQ ID NO: 15) to create a new molecule "ADT1-4-2 (L74S)". The affinity of these molecules (relative to the starting molecules ADT1-4 and ADT1-4-2, respectively) was determined by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human or cynomolgus vδ1 heterodimers flowed through cells at concentrations of 100nM, 50nM, 25nM, 12.5nM, 6.25nM with the following parameters: 180 seconds association, 480 seconds dissociation, flow rate 25. Mu.L/min, running buffer PBS+0.05% Tween 20. All experiments were performed at room temperature: (D i) the resulting SPR trace depicts the affinity of the indicated antibodies for human and cynomolgus vδ1 antigen and a tabulated summary of (D ii) results.

Fig. 27: CDR3 usage and cross sharing. (A) Examples of cross sharing of heavy and light chains between affinity matured antibodies in ADT1-4 (G04) lineage (0.4 nM human TRDV ELISA results heat-map fractionation). This "heat map" demonstrates the cross sharing of CDR3 sequences between different affinity matured antibodies in the ADT1-4 lineage, showing that the antibody sequences provided by the affinity maturation process need not be provided as specific heavy and light chain pairs. The results underscored that affinity matured antibodies can cross-share light and heavy chains, and that different LC/HC combinations produce equivalent or improved results in high stringency antigen binding studies. ADT1-4 (G04) parental clones (bottom left) are also included. (B) ADT1-4 lineage final selection: CDR3 was used and cross-shared + cynomolgus monkey antigen binding 'heatmap' with the starting parent ADT1-4 parent G04 mAb (bottom left). (C) ADT1-7 lineage final selection: CDR3 was used and cross-shared + human antigen binding 'heatmaps' with the starting ADT1-7 parent E07 mAb (bottom left).

Fig. 28: fold enhancement of ADT1-4 lineage binding compared to ADT1-4 parent G04. (A) fold enhancement of binding of recombinant human V.delta.1 antigen. (B) fold enhancement of binding of recombinant cynomolgus monkey vδ1 antigen. (C) enhancement fold of primary V.delta.1 MFI. (D) fold enhancement of MFI in PEER V.delta.1 cell lines.

Fig. 29: fold enhancement of ADT1-7 lineage binding compared to ADT1-7 parent E07. (A) fold enhancement of binding of recombinant human V.delta.1 antigen. Enhancement fold of primary vδ1 MFI. Enhancement fold of MFI for the PEER vδ1 cell line.

Figure 30 fold improvement in human (and cynomolgus monkey) antigen binding compared to parental clones. (A) fold improvement in binding to human antigen. Affinity matured clones were signaled DELFIA ELISA to the parental ADT1-4 (G04), 0.4nM recombinant human L1 antigen (containing human vδ1 TCR antigen). (B) Fold improvement in binding to cynomolgus monkey antigen (DV 1/GV77 containing cynomolgus monkey SEQ ID NO:308 (mature, minus leader sequence). DELFIA ELISA signals of affinity matured clones with parental ADT1-4 (G04), 0.4nM recombinant human L1 antigen. (C) fold improvement in binding to human antigen. DELFIA ELISA signals of affinity matured clones with parental ADT1-7 (E07), 0.4nM recombinant human L1 antigen.

FIG. 31 fold improvement in KD for human (and cynomolgus monkey) antigens compared to parental clones. (A) Fold improvement in KD of ADT1-4 (G04) lineage to human antigen. (B) Fold improvement in KD of ADT1-4 (G04) lineage against cynomolgus monkey antigen. (C) Fold improvement of the ADT1-7 (E07) lineage on human antigen KD.

FIG. 32 binding affinity assay (KD, by SPR) for human V.delta.1 antigen. (A) surface plasmon resonance of ADT1-4 lineage. (B) surface plasmon resonance of ADT1-7 lineage. (C) KD values for ADT1-4 lineages and fold change relative to parental clones. (D) KD values for ADT1-7 lineages and fold changes relative to parental clones. (E) Fold change in KD of ADT1-4 lineage relative to parental clones. (F) Fold change in KD of ADT1-7 lineage relative to parental clones.

FIG. 33 binding affinity assay to cynomolgus monkey antigen (KD, by SPR). (A) Surface plasmon resonance of the ADT1-4 lineage on cynomolgus monkey antigens. (B) KD value of ADT1-4 lineage to cynomolgus monkey antigen.

FIG. 34 binding affinity to cell surface V.delta.1 TCR (EC 50 for binding to cell surface V.delta.1). Binding levels of (A, B) vδ1 mAb to two γδ T cell donors ATS006 (a) and TS164 (B). (C) Bars represent the average 50% binding values for ADT1-4 and ADT1-7 clones to vδ1-positive γδ T cells, expressed as an average of two donors, provided in the table as% improvement. (D) The table summarizes the IC50 plotted in (a) and (B), as well as the vδ1 negative cell types, including HEK293A, raji cells and various leukocyte subpopulations within primary blood mononuclear cells. For vδ1 positive γδ T cells, the data are expressed as the average of two donors.

Tcr down-regulation (using soluble antibodies) fig. 35. (A, B) ADT1-4 lineage (A) and ADT1-7 lineage (B). (C) Average TCR down-regulation-IC 50 results from two GD cell donors. (D, E) TCR downregulation fold improvement over the parental ADT1-4 clone (D) and ADT1-7 clone (E). (F) 50% effect value from (C), wherein the percentage improvement is calculated based on the parents of each of ADT1-4 (up) and ADT1-7 (down). (G) 50% effect value from (C), and fold improvement calculated from the parents of each of ADT1-4 (up) and ADT1-7 (down).

FIG. 36 effects of V.delta.1 monoclonal antibodies on gamma.delta.activation as measured by CD107a expression. (A, B) cloning of ADT1-4-2 in GD cells either alone (A) or together with THP-1 cells (B). (C, D) cloning of ADT1-7-3 in GD cells alone (C) or together with THP-1 cells (D). (E) Table shows the percentage increase in γδ CD107a expression of co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, non-co-cultured γδ cells. (F) Table shows the percentage increase in γδ CD107a expression for co-cultured cells treated with the highest concentration vδ1 mAb compared to untreated, co-cultured and non-co-cultured γδ cells.

FIG. 37 effect of V.delta.1 monoclonal antibodies on gamma.delta.activation as measured by CD25 expression. (A, B) cloning of ADT1-4-2 in cell GD cells alone (A) or together with THP-1 cells (B). (C, D) cloning of ADT1-7-3 in cell GD cells alone (C) or together with THP-1 cells (D). (E) Table shows the percentage increase in γδ CD25 expression for co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, non-co-cultured γδ cells. (F) Table shows the percentage increase in γδ CD25 expression for co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, co-cultured and non-co-cultured γδ cells.

Fig. 38 cynomolgus TCR down-regulation (using soluble antibodies). (A) Comparison of the ability of ADT1-4-2 and ADT1-4 to bind VD1 on cynomolgus monkey γδ -T cells and reduce their expression. (B) Percent cell surface expression of VD1 after ADT1-4-2 treatment. (C) Individual EC50 values for different donors were combined with mean and standard deviation. (D-G) other studies were performed on cynomolgus monkey whole blood samples (n=5).

FIG. 39. Quantification of viable THP-1 cells after 24 hours of co-culture with γδT cells in the presence of V.DELTA.1 mAb or control. (A) THP-1 cell killing assay of ADT1-4 clone. (B) THP-1 cell killing assay of ADT1-7 clone. (C) Average EC50 in THP-1 killing assay of ADT1-4 and ADT1-7 clones. (D) Table summarizes the EC50 s plotted in (C).

FIG. 40. Influence of V.delta.1 monoclonal antibodies on Antibody Dependent Cellular Cytotoxicity (ADCC).

FIG. 41. Influence of V.delta.1 monoclonal antibodies on Complement Dependent Cytotoxicity (CDC).

FIG. 42 healthy cell retention of clone ADT 1-4-2.

FIG. 43 effect of anti-V.delta.1 antibodies on TIL populations of primary tumor biopsies. (A, B) shows a decrease in vδ1 TCR expression on total tumor infiltrating γδ T cells after mAb stimulation 48 (a) or 72 (B) hours in two independent donors confirming target engagement in each case. (C) Shows enhanced expression of CD25 and Ki67 on V.delta.1+ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or ADT1-4 stimulation. (D) It was shown that IFN-. Gamma.production of TIL was significantly increased by stimulation with ADT1-4-2 or ADT1-7-3 in the presence of 50ng/ml IL-15 for 72 hours. (E) It was shown that stimulation of TIL with ADT-1-4-2 or ADT1-7-3 at this time point did not enhance secretion of the type 17 related cytokines IL-6 or IL-17. (F, G) showed a decrease in vδ1 TCR expression on total tumor infiltrating γδ T cells 24 (F) or 72 (G) hours after mAb stimulation of two individual donors confirming target engagement in TIL isolated by enzymatic digestion. (H) Shows a dose-dependent increase in Ki67 expression on γδ T cells after 72 hours of stimulation with ADT 1-4-2. (I, J) shows fold increase in IFN-gamma production by enzyme digestion at 24 (I) or 72 (J) hours from two individual donors and TIL stimulated with ADT1-4-2 at a concentration of 6.66nM in the presence of 2ng/ml IL-15. In all cases, the concentrations of control anti-RSV and parent ADT1-4 matched the highest concentrations used in the study (i.e., A, C, F, G, H, I, J at 6.66nM, and 66.6nM for B and E).

FIG. 44 effects of anti-V.delta.1 antibodies on TIL populations of primary tumor biopsies. (A) Shows enhanced expression of CD25 and Ki67 in γδ T cells stimulated with ADT1-4-2 for 10 days. (B) It was shown that perforin ⁺ granzyme B ⁺ γδ T cells were greatly increased after 10 days of stimulation with ADT 1-4-2. (C) It was shown that granzyme B and perforin expression of cd8+ and CD8- αβ T cells was significantly enhanced after 10 days of tumor infiltration vδ ⁺ T cells stimulated with ADT 1-4-2. (D) Shows a significant increase in IFN-gamma production of lung tumor derived TIL and a modest increase in IL-17 and IL-6 production following stimulation with ADT 1-4-2. I demonstrates an enhancement in production of the chemokines CCL2, CCL4 and CXCL10 by TIL after 10 days of stimulation with ADT 1-4-2. In all cases, the control anti-RSV and parental ADT1-4 matched the same concentration as affinity matured ADT 1-4-2.

Fig. 45: sequences of ADT1-4 lineage clones (light chain)

Fig. 46: sequences (heavy chain) cloned from ADT1-4 lineage.

Fig. 47: sequences cloned in ADT1-7 lineage (light chain).

Fig. 48: sequences (heavy chain) of ADT1-7 lineage clones

Fig. 49: the anti-vδ1/CD19 bispecific antibodies showed high affinity binding to human vδ1 and cynomolgus vδ1, comparable to the parent monoclonal vδ1 mAb. Surface Plasmon Resonance (SPR) analysis was performed on ADT1-4-2/CD19 bispecific antibodies to assess binding to human V.delta.1 and CD19 antigens. Bispecific antibodies bind both human and cynomolgus vδ1 with only a slight decrease in affinity for both antigens.

Fig. 50: the vδ1-CD19 bispecific T cell cement enhances cytotoxicity and γδ T cell activation of CD19 ⁺ target cells while retaining healthy CD19 ⁺ B cells. (A) The expression of CD19 on cancerous NALM-6, raji, primary B cells and V.delta.1γδ T cells was determined by flow cytometry. (B-D) Effect of the parent (ADT 1-4) and affinity maturation (ADT 1-4-2) V.delta.1-CD 19 bispecific antibodies on cytotoxicity of NALM-6 cells (B), raji cells (C) and B cells (D). Antibodies were titrated with vδ1γδ T cells in the presence of a 1:1 e:t ratio for 12 hours. The percentage of viable cells was calculated by high content confocal microscopy and normalized to viable cell count in the absence of vδ1γδ T cells. (E) Bars represent the percentage of viable cells after 12 hours of co-culture of vδ1γδ T cells (NALM-6, raji or B cells) in the presence of vδ1 bispecific antibodies and controls, as shown in (B), (C) and (D). (F-G) the effect of the V.delta.1-CD 19 bispecific antibody on V.delta.1 TCR surface expression in the presence of NALM-6 target cells (F) and healthy B cells (G) was determined by flow cytometry after 4 hours of co-culture. (H) Bar graph represents the maximum TCR down-regulation percentage at the highest concentration (3 μg/ml), as shown in (F) and (G). (I-K) the effect of a Bv.delta.1-CD 19 bispecific antibody on the up-regulation of CD107a on the surface of Bv.delta.1 cells in the presence of NALM-6 target cells (I) and healthy B cells (J) was determined by flow cytometry after four hours of co-culture. All data represent mean ± standard deviation, and represent n=3. Bleb mab (CD 3-CD19 BiTE) was included as a control in all assays.

Fig. 51: affinity matured vδ1 clones in dual-specific format bind to Her2 ⁺ target cells and exhibit enhanced binding to vδ1γδ T cells, as well as enhanced cytotoxicity of Her2 ⁺ target cells. (A) Cell surface expression of Her2 and vδ1 on breast cancer cell lines and vδ1γδt cells (B). Binding of (C-F) V.delta.1-Her 2 bispecific antibody and Her2 mAb control (Trastuzumab) to Her2+ (SK-BR-3 (C), BT-474 (D)), her2- (MDA-MD-231 (E)) and V.delta.1 ⁺ cells (F). (G-I) percentage of viable cells remaining after 24 hours of co-culture with V.delta.1-Her 2 bispecific antibody at a 1:1E:T ratio with V.delta.1γδ T cells and SK-BR-3 cells (G), BT-474 (H) and MDA-MB-231 cells (I). (J) Bar graphs show the percent increase in cytotoxicity of vδ1γδ T cell vδ1-Her2 bispecific antibody after 24 hours.

Fig. 52: the vδ1/EGFR bispecific antibodies exhibit high affinity binding to human EGFR and human vδ1 binding affinity comparable to their parent mAb. (A, B) Surface Plasmon Resonance (SPR) analysis was performed on vδ1/EGFR bispecific antibodies to assess binding to human vδ1 (a) and EGFR antigen (B). Parent mAb, cetuximab, and negative control mAb were included for comparison purposes.

Fig. 53: the vδ1/EGFR bispecific antibody binds to EGFR ⁺ a431 target cells and vδ1γδ T cells. (A) Cell surface expression of EGFR and vδ1 on a431 cell line and primary vδ1γδ T cells. (B) Binding levels of vδ1/EGFR bispecific antibodies to a431 cell line or primary vδ1γδ T cells. Target cells were stained with different concentrations of antibody followed by fluorescent anti-human IgG detection antibody. All incubation steps were performed at 4 ℃ and mAb binding was determined using flow cytometry to measure median fluorescence levels. GRAPHPAD PRISM 9 was used to fit a log-four parameter dose-response curve.

Fig. 54: the vδ1/EGFR bispecific antibody induces EGFR-specific T cell activation and degranulation, resulting in γδ T cell mediated increase in cytotoxicity of a431 target cells. (A) Cell surface expression of γδ TCR on primary vδ1γδ T cells after 24 hours of incubation with bispecific antibody in the presence or absence of a431 cells. (B, C) number of viable A431 cells after 24 hours of co-culture at a 1:1 ratio and different concentrations of antibody (B) and activation status of primary V.delta.1γ.delta.T cells (C). Viability was measured by viability dye and activation status using CD25 antibodies. (D) Degranulation of primary V.delta.1γ.delta.T cells after four hours of co-culture with A431 cells at a 1:1 ratio and varying concentrations of antibody. Degranulation was determined by adding fluorophore conjugated anti-CD 107 a antibodies directly to the cell-antibody mixture at the beginning of co-culture. (E) Number of viable a431 cells after 24 hours co-culture with 10pM antibody and varying amounts of primary vδ1γδ T cells. (A-E) in each case, fluorescence was determined using flow cytometry to measure median fluorescence levels. GRAPHPAD PRISM 9 was used to fit a log-four parameter dose-response curve. Data are expressed as mean ± SD of two biological replicates.

Fig. 55: further evidence of non-depletion and down-regulation of CD3 in blood-derived and tumor-associated vδ1T cells. (A) V delta 1 TCR MFI following antibody stimulation as an indication of mAb target engagement is shown. (B) MFI of CD3 expression on positively gated blood-derived vδ1 cells is shown. The use of vδ1 antibodies stimulated the engagement of vδ1 cells and caused the down-regulation of vδ1 and CD3 on vδ1 cells. (C, D and E) show that exemplary anti-V.delta.1 antibodies of the invention confer activation and non-depletion effects on tumor-associated V.delta.1T cells.

Fig. 56: vδ1×fapα bispecific antibodies enhance vδ1γδ T cell activation and fapα ⁺ fibroblast lysis. (A) The binding kinetics of anti-vδ1 (ADT 1-4-2), anti-fapα monoclonal (based on sibutrab (Sibrotuzumab)) and anti-vδ1×fapα (ADT 1-4-2×sibutrab) bispecific antibodies to recombinant human vδ1 and fapα are shown, as determined by Surface Plasmon Resonance (SPR). (B, C) binding of anti-vδ1 and anti-fapα antibodies to fapα ⁺ fibroblasts (B) and vδ1γδ T cells (C). (D, E) effect of anti-vδ1×fapα bispecific antibodies and monoclonal controls on vδ1 TCR down-regulation on vδ1γδ T cells in the absence (D) or presence of fapα ⁺ fibroblasts (E). The effect of (F-G) anti-vδ1×fapα bispecific antibodies and monoclonal controls on CD107a upregulation on vδ1γδ T cells in the absence (F) or presence of fapα ⁺ fibroblasts (G). (H) Effects of anti-vδ1×fapα bispecific antibodies and monoclonal controls on V δ1γδ T cell induced fibroblast lysis after 24 hours of co-culture. The percent viable cells were calculated by high content confocal microscopy and normalized to viable cell count in the absence of vδ1γδ T cells.

Fig. 57: vδ1×msln bispecific antibodies enhance vδ1γδ T cell activation and MSLN ⁺ target cell lysis. (A) Binding kinetics of anti-vδ1 (ADT-1-4-2), anti-MSLN monoclonal (based on antibodies disclosed in US 2014/0004121) and anti-vδ1×msln (ADT 1-4-2×msln) bispecific antibodies to recombinant human vδ1 and MSLN are shown, as determined by Surface Plasmon Resonance (SPR). (B, C) binding of anti-vδ1 and anti-MSLN antibodies to MSLN ⁺ HeLa cells (B) and vδ1γδ T cells (C). (D, E) effect of anti-vδ1×msln bispecific and monoclonal antibodies on vδ1 TCR down-regulation on vδ1γδ T cells in the absence (D) or presence of MSLN ⁺ OVCAR-3 cells (E). (F, G) effect of anti-vδ1×msln bispecific antibodies and monoclonal controls on CD107a upregulation on vδ1γδ T cells in the absence (F) or presence of MSLN ⁺ OVCAR-3 cells (G). (H) Effects of anti-vδ1×msln bispecific antibody and monoclonal control on HeLa cell lysis by vδ1γδ T cells after 24 hours of co-culture. The percent viable cells were calculated by high content confocal microscopy and normalized to viable cell count in the absence of vδ1γδ T cells.

Fig. 58: the vδ1×pd-1 bispecific antibody enhances activation of vδ1γδ T cells and inhibits checkpoint blockade of PD-1 ⁺ T cells. A) SPR analysis of anti-V.delta.1 (ADT 1-4-2), anti-PD-1 (based on palbockizumab), anti-RSVIgG control X anti-PD-1 and anti-V.delta.1X anti-PD-1 (ADT 1-4-2X palbockizumab) bispecific antibodies binding to recombinant human V.delta.1 and PD-1. B) Dual binding of anti-vδ1 and anti-PD-1 bispecific antibodies to recombinant human PD-1 and vδ1 as determined by SPR. C) Expression of PD-1 on CD4 and CD 8T cells activated with anti-CD 3/anti-CD 28 dynabeads. D) Binding of anti-vδ1 and anti-PD-1 antibodies to PD-1+ activated CD4 and CD 8T cells and vδ1γδ T cells. E) The effect of anti-vδ1×pd-1 bispecific antibodies and monoclonal controls on vδ1 TCR down-regulation on vδ1γδt cells in the presence or absence of PD-1 ⁺ CD 4T cells. F) EC50 of anti-vδ1×pd-1 bispecific antibodies down-regulates vδ1 TCR on vδ1γδt cells in the presence or absence of PD-1 ⁺ CD 4T cells. G) Effects of vδ1 cross-linked anti-vδ1×pd-1 bispecific antibodies on PD-1 ⁺ T cell activation.

Fig. 59: the vδ1x4-1 BB bispecific antibody enhances activation of vδ1γδ T cells and 4-1BB ⁺ T cells. A) SPR analysis of binding of anti-V.delta.1 (ADT 1-4-2), anti-4-1 BB (based on Wu Tuolu mab (utomilumab)), anti-RSVIgG control X anti-4-1 BB and anti-V.delta.1X anti-4-1 BB (ADT 1-4-2X Wu Tuolu mab) bispecific antibodies to recombinant human V.delta.1 and 4-1 BB. B) The anti-vδ1 and anti-4-1 BB bispecific antibodies as determined by SPR double bind to recombinant human 4-1BB and vδ1. C) Expression of 4-1BB on CD4 and CD 8T cells activated with anti-CD 3/anti-CD 28 dynabeads. D) Binding of anti-vδ1 and anti-4-1 BB antibodies to 4-1BB ⁺ activates CD 8T cells and vδ1γδ T cells. E-F) effect of anti-vδ1x4-1 BB bispecific antibody and monoclonal control on vδ1γδt cell down-regulation of vδ TCR in the absence (E) or presence of 4-1BB ⁺ CD 8T cells (F). G) Effect of V.delta.1 Cross-Linked anti-V.delta.1X 4-1BB bispecific antibodies on 4-1BB ⁺ T cell activation.

Fig. 60: the vδ1xox 40 bispecific antibody enhances activation of vδ1γδ T cells and OX40 ⁺ T cells. A) SPR analysis of anti-vδ1 (ADT 1-4-2), anti-OX 40 (based on pergolizumab (pogalizumab)), anti-RSVIgG control x anti-OX 40 and anti-vδ1 x anti-OX 40 (ADT 1-4-2 x pergolizumab) bispecific antibodies binding to recombinant human vδ1 and OX 40. B) Dual binding of anti-vδ1 and anti-OX 40 bispecific antibodies to recombinant human OX40 and vδ1 as determined by SPR. C) Expression of OX40 on CD4 and CD 8T cells activated with anti-CD 3/anti-CD 28 dynabeads. D) Binding of anti-vδ1 and anti-OX 40 antibodies to OX40 ⁺ activated CD 4T cells and vδ1γδ T cells. E-F) effect of anti-vδ1xox 40 bispecific antibodies and monoclonal controls on vδ TCR down-regulation on vδ1γδ T cells in the absence (E) or presence of OX40 ⁺ CD 4T cells (F). G) Effects of vδ1 cross-linked anti-vδ1×ox40 bispecific antibodies on OX40 ⁺ T cell activation.

Fig. 61: vδ1×tigit bispecific antibodies enhance activation of vδ1γδ T cells and inhibit checkpoint blockade of TIGIT ⁺ T cells. A) SPR analysis of binding of anti-V.delta.1 (ADT 1-4-2), anti-TIGIT (based on Tiarella Li Youshan anti (tiragolumab)), anti-RSVIgG control X anti-TIGIT and anti-V.delta.1X anti-TIGIT (ADT-1-4-2X Tiarella Li Youshan anti) bispecific antibodies to recombinant human V.delta.1 and TIGIT. B) The dual binding of anti-vδ1 and anti-TIGIT bispecific antibodies to recombinant human TIGIT and vδ1 as determined by SPR. C) TIGIT expression on CD4 and CD 8T cells activated with anti-CD 3/anti-CD 28 dynabeads. D) Binding of anti-vδ1 and anti-TIGIT antibodies to TIGIT ⁺ activates CD4 and CD 8T cells and vδ1γδ T cells. E) The effect of anti-vδ1×tigit bispecific antibodies and monoclonal controls on vδ1 TCR down-regulation on vδ1γδ T cells with or without TIGIT ⁺ CD 8T cells. F) EC50 of anti-vδ1×tigit bispecific antibodies down-regulate vδ1 TCR on vδ1γδt cells with or without TIGIT ⁺ CD 8T cells. G) Effect of V.delta.1 Cross-Linked anti-V.delta.1×TIGIT bispecific antibodies on TIGIT ⁺ T cell activation.

Fig. 62: ADCC reporter bioassays did not show ADCC caused by anti-vδ1 antibodies. Target cells, γδ cells, were incubated with ADCC bioassay effector cells in the presence of anti-vδ1 antibodies, anti-vδ1LAGA antibodies (Fc disabled) and RSV isotype controls. Luminescence signals were recorded as Relative Light Units (RLU) and fold induction was calculated as described in the methods. The γδ donor n=2 of "anti-vδ1 antibody", "anti-v 61 LAGA antibody", "RSV", "OKT3" (performed in technical duplicate). The Raji cell line n=1 for the "rituximab+raji" condition (in technical replicates) and the γδ donor n=1 for the "anti-vδ1 antibody+effector" and "anti-vδ1LAGA antibody+effector" conditions (in technical replicates and single runs, respectively). The ratio of effector cells to target cells was 3:1.

Fig. 63: the vδ1-CD19 bispecific T cell cement enhances cd19+ target cell cytotoxicity and γδ T cell activation while retaining healthy cd19+ B cells. a-F) effects of anti-vδ1×cd19 and cd3×cd19 bispecific antibodies on γδ T cells or αβ T cell mediated Raji cells or healthy primary B cell lysis. The percentage of positive Raji or healthy primary B cells was determined by confocal microscopy at 24 hours in the following: ternary cultures of vδ1γδ T cells with Raji cells (a) and primary healthy B cells (B); ternary cultures of αβγt cells with Raji cells (C) and primary healthy B cells (D); or quaternary cultures of vδ1γδ T cells and αβ T cells with Raji cells (E) and primary healthy B cells (F). G-I) quantification of IL-17A secreted by γδT cells or αβT cells 24 hours after stimulation with anti-vδ1xCD19 and CD3xCD19 bispecific antibodies. Cell culture supernatants from co-cultures of Raji cells, primary B cells, and the following were collected: γδ T cells (G); or αβ T cells (H); or γδ T cells and αβ T cells (I), and IL-17A secretion is determined by MSD. The dashed line represents the lowest quantitative level.

Fig. 64: stimulation with high affinity anti-vδ1+ antibodies enhanced expression of 4-1BB on vδ1γδ T cells. (A) Enhancement of 4-1BB expression by addition of exemplary high affinity anti-V.delta.1 antibodies: tissue-derived V.delta.1 ⁺ T cells from 4 different human donors were incubated with the indicated antibodies (1. Mu.g/ml) in the presence of THP-1 cells (2:1E:T). After 4 hours, expression of 4-1BB on the surface of V.delta.1 ⁺ T cells was analyzed by flow cytometry. At this time point, cells stimulated with ADT1-4-2 significantly increased 4-1BB expression, comparable to stimulation with OKT3 (< 0.05, < 0.01, < p), as determined by common one-way anova with Sidak post hoc testing. (B, C) enhancement of 4-1BB expression; further study: tissue-derived V.delta.1 ⁺ T cells were incubated with a drop of the indicated ADT1-4-2 or anti-IgG 1 isotype control at a 1:1 ratio in the presence of (B) THP-1 cancer cells or (C) healthy blood-derived monocytes. After 4 hours, expression of 4-1BB on the surface of V.delta.1 ⁺ T cells was analyzed by flow cytometry. Stimulation with ADT1-4-2 increased expression of 4-1BB on the surface of V.delta.1T cells. In addition, even though activated vδ1 ⁺ cells retain healthy cells (see further discussion elsewhere herein), similar antibody-induced activation was observed when tumor or healthy fcγr ⁺ cells provided cross-linking.

Fig. 65: upregulation of Native Cytotoxic Receptor (NCR) on vδ1+ T cells in primary PBMCs (a) experimental culture setup, including validated TCR downregulation assays: human PBMC cultures from three different donors were incubated for 10 days with exemplary affinity matured ADT1-4 and ADT1-7 anti-V.delta.1 antibody clones (see also B, C and D) or anti-RSV controls shown at 3.33 nM. Cultures were also supplemented with 4ng/ml IL-2 and 10ng/ml IL-15 (Miltenyi, 130-097-746 and Peprotech,200-15-500ug, respectively). After 10 days of culture, vδ1+ T cell% was analyzed by flow cytometry. All cultures incubated with anti-vδ1 antibodies showed a significant decrease in detectable vδ1+ cells (in% pan gamma) relative to anti-RSV control culture j. Representative and typical results for one such antibody are shown herein. From this analysis, it was confirmed that all affinity-matured anti-vδ1 antibodies included in this study exhibited significant target binding and conferred TCR down-regulation. (B-D) anti-vδ1 antibodies enhance the expression of NCR (NKp 30, NKp44, NKp 46) on detectable vδ1+ cells: in addition to confirming target engagement of all anti-vδ1 antibodies used in this study, fold changes in NCR were studied on gated vδ1+ cells. Specifically, (B) exhibits a fold change in biscationic vδ1+, nkp30+% relative to the RSV control (RSV control normalized on the far right and expressed as 1-fold). (C) Fold changes were presented for biscationic vδ1+, nkp44+% relative to RSV control (RSV control normalized on the far right and expressed as 1 fold). (D) Fold changes were presented for biscationic vδ1+, nkp46+% relative to RSV control (RSV control normalized on the far right and expressed as 1 fold). These combined results underscore the consistent upregulation of NCR (donor dependent) in detectable vδ1+ conferred by all affinity matured anti-vδ1+ antibodies used in this extended 10 day primary PBMC culture system.

Detailed Description

The present invention provides high affinity anti-TCR delta variable 1 (anti-vδ1) antibodies, multispecific antibodies, and antibody fragments thereof. More specifically, the invention relates to optimizing antibodies, e.g., antibodies prepared starting from a parent anti-vδ1 antibody according to an optimization selection procedure, e.g., the provision and characterization of parent antibodies referred to herein as G04, E07, C08, B07, C05, E04, F07, G06, G09, B09, G10, and E01. The invention relates in particular to optimised antibodies derived from G04 and E07.

Definition of the definition

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the following terms have the meanings ascribed to them below.

Γδ (GAMMA DELTA) T cells represent a small subset of T cells that express a unique, defined T Cell Receptor (TCR) on the surface. The TCR consists of a gamma (gamma) chain and a delta (delta) chain. Each chain contains a variable (V) region, a constant (C) region, a transmembrane region and a cytoplasmic tail. The V region contains an antigen binding site. There are two main subtypes of human γδ T cells: a subtype that predominates in peripheral blood and a subtype that predominates in non-hematopoietic tissue. Both subtypes may be defined by the type of delta and/or gamma present on the cell. For example, γδt cells, which predominate in peripheral blood, express mainly δ variable 2 chains (vδ2). γδ T cells that predominate in non-hematopoietic tissues (i.e., tissue resides) express predominantly δ variable 1 chains. Reference to "vδ1T cells" or "vδ1+t cells" refers to γδt cells having a vδ1 chain, i.e. vδ1 ⁺ cells.

Reference to "delta variable 1" may also be referred to as vδ1 or Vd1, and the nucleotide encoding the TCR chain containing the region or the TCR protein complex comprising the region may be referred to as "TRDV1". The antibody or antigen-binding fragment thereof that interacts with the vδ1 chain of the γδ TCR is virtually all an antibody or antigen-binding fragment thereof that binds vδ1 and may be referred to as an "anti-TCR δ variable 1 antibody or antigen-binding fragment thereof" or an "anti-vδ1 antibody or antigen-binding fragment thereof" or an "anti-TRDV 1 antibody or antigen-binding fragment thereof.

Additional references are made herein to other delta chains, such as "delta variable 2" chains. These can be cited in a similar manner. For example, a delta variable 2 chain may be referred to as vδ2, while a nucleotide encoding a TCR chain containing or a TCR protein complex comprising that region may be referred to as "TRDV2". In a preferred embodiment, the antibody or antigen binding fragment thereof that interacts with the vδ1 chain of the γδ TCR does not interact with other δ chains such as vδ2. In the present invention, the antibody is specific for TRDV1 and does not bind to TRDV (SEQ ID NO: 310) or other antigens present on the γδ T cell receptor, such as TRDV (SEQ ID NO: 311).

Also referred to herein are "gamma variable chains". These may be referred to as gamma-chains or V gamma, and the nucleotide encoding the TCR chain containing the region or the TCR protein complex comprising the region may be referred to as TRGV. For example TRGV4 refers to the vγ4 chain. In a preferred embodiment, the antibody or antigen binding fragment thereof that interacts with the V.delta.1 chain of the gamma.delta.TCR does not interact with a gamma chain such as V.gamma.4 (e.g., SEQ ID NO: 309). Antibodies also do not bind or interact with other domains found in γδ TCRs (e.g., TRDJ, TRDC, TRGJ or TRGC).

The term "T cell receptor complex" is a complex comprising proteins found on the surface of T cells that are responsible for the recognition of "T cell receptors" (or "TCRs") of various antigens. Although the exact composition of the T cell receptor complex may vary, the T cell receptor complex comprises the alpha and beta chains of the T cell receptor, or in the case of γδ T cells, the γ and δ chains of the T cell receptor, and up to 6 or more additional chains, such as cd3δ, cd3γ, cd3ε, and cd3ζ. The T cell receptor complex mediates intracellular signaling in T cells, which can lead to T cell activation.

The term "antibody" includes any antibody protein construct comprising at least one antibody variable domain comprising at least one Antigen Binding Site (ABS). Antibodies include, but are not limited to, immunoglobulins of the type IgA, igG, igE, igD, igM (and subtypes thereof). The overall structure of an immunoglobulin G (IgG) antibody assembled from two identical heavy (H) chains and two identical light (L) chain polypeptides is well established and highly conserved in mammals (Padlan (1994) mol. Immunol. 31:169-217).

Conventional antibodies or immunoglobulins (Ig) are proteins comprising the following four polypeptide chains: two heavy (H) chains and two light (L) chains. Each chain is divided into a constant region and a variable domain. The heavy (H) chain variable domain is abbreviated herein as VH and the light (L) chain variable domain is abbreviated herein as VL. These domains, domains related to these domains, and domains derived from these domains may be referred to herein as immunoglobulin chain variable domains. VH and VL domains (also known as VH and VL regions) can be further subdivided into regions termed "complementarity determining regions" ("CDRs") interspersed with regions that are more conserved (termed "framework regions" ("FR")). The framework regions and complementarity determining regions have been precisely defined (Kabat et al Sequences of Proteins of Immunological Interest, fifth edition U.S. Department of Health and Human Services, (1991) NIH publication No. 91-3242). Alternative numbering conventions for CDR sequences also exist, such as Chothia et al (1989) Nature 342:877-883 or those outlined in imgt. In conventional antibodies, each VH and VL consists of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Conventional antibody tetramers of two immunoglobulin heavy chains and two immunoglobulin light chains are formed from immunoglobulin heavy chains and immunoglobulin light chains that are interconnected by, for example, disulfide bonds, and the heavy chains are similarly linked. The heavy chain constant region comprises three domains, CH1, CH2 and CH3. The light chain constant region comprises one domain CL. The variable domains of the heavy and light chains are binding domains that interact with an antigen. The antibody constant region generally mediates binding of the antibody to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1 q).

As used herein, a "fragment" of an antibody (which may also be referred to as an "antibody fragment," "immunoglobulin fragment," "antigen binding fragment," or "antigen binding polypeptide") refers to a portion of an antibody (or construct containing such a portion) that specifically binds to a target, i.e., a delta variable 1 (vδ1) chain of γδ T cell receptor (e.g., a molecule in which one or more immunoglobulin chains are not full length but specifically bind to the target). Examples of binding fragments encompassed within the term antibody fragment include:

(i) Fab fragments (monovalent fragments consisting of VL, VH, CL and CH1 domains);

(ii) F (ab') 2 fragments (bivalent fragments consisting of two Fab fragments linked at the hinge region by a disulfide bridge);

(iii) Fd fragment (consisting of VH and CH1 domains);

(iv) Fv fragment (consisting of VL and VH domains of the antibody single arm);

(v) A single chain variable fragment scFv (consisting of a VL domain and a VH domain joined by a synthetic linker using recombinant methods, the synthetic linker enabling the VL domain and VH domain to form a single protein chain, wherein the VL region and VH region pair to form a monovalent molecule);

(vi) VH (immunoglobulin chain variable domain consisting of VH domains);

(vii) VL (immunoglobulin chain variable domain consisting of VL domains);

(viii) Domain antibodies (dabs, consisting of VH or VL domains);

(ix) Minibodies (consisting of a pair of scFv fragments linked via a CH3 domain); and

(X) Diabodies (consisting of non-covalent dimers of scFv fragments consisting of VH domains from one antibody linked by small peptide linkers to VL domains from another antibody).

"Human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. The human subject to whom the human antibody is administered does not develop a cross-species antibody response (e.g., known as HAMA response-human anti-mouse antibody) to the primary amino acids contained within the antibody. The human antibodies may include amino acid residues in the CDRs, and in particular in CDR3, that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis or by somatic mutation). However, the term is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted onto human framework sequences. Human antibodies are prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant combinatorial human antibody libraries, antibodies isolated from animals (e.g., mice) that are transgenic for human immunoglobulin genes, or antibodies prepared, expressed, created or isolated by any other means that involves the splicing of human immunoglobulin gene sequences to other DNA sequences, also referred to as "recombinant human antibodies".

Substitution of at least one amino acid residue in the framework region of a non-human immunoglobulin variable domain with a corresponding residue from a human variable domain is referred to as "humanization". Humanization of the variable domains may reduce immunogenicity in humans.

"Specificity" refers to the number of different types of antigens or antigenic determinants to which a particular antibody or antigen-binding fragment thereof can bind. The specificity of an antibody is the ability of an antibody to recognize a particular antigen as a unique molecular entity and to distinguish it from other antigens. Antibodies that "specifically bind" to an antigen or epitope are terms well known in the art. A molecule is said to exhibit "specific binding" if it reacts more frequently, more rapidly, longer in duration, and/or with higher affinity to a particular target antigen or epitope than it reacts to an alternative target. An antibody "specifically binds" to a target antigen or epitope if it binds to the target antigen or epitope with greater affinity, higher avidity, easier and/or longer duration than it binds to other substances.

Antibodies of the invention include monospecific antibodies (i.e., antibodies that bind only one antigen) and multispecific antibodies. A "multispecific antibody" is an antibody capable of binding to multiple different epitopes simultaneously or sequentially. Typically, the epitopes are not on the same antigen. Thus, multispecific antibodies have the ability to selectively bind to epitopes present on different antigens via a plurality of different binding domains. This is in contrast to conventional monospecific antibodies that do not have this capability. In contrast, "monospecific antibodies" have binding specificity for only one antigen, although they may have multiple binding sites for that antigen (e.g., a fully human IgG antibody may have a valence of 2 and other antibodies may have higher valence, if an antibody recognizes only one antigen, it is still classified as a monospecific antibody). Thus, the multispecific antibodies of the present invention bind to a plurality of different antigens simultaneously and/or sequentially.

In some embodiments of the invention, the antibody is a bispecific antibody. A "bispecific antibody" is an antibody capable of binding two different epitopes simultaneously and/or sequentially. Typically, the epitopes are not on the same antigen. Thus, bispecific antibodies have the ability to selectively bind to two different epitopes present on two different antigens via two different binding domains. This is in contrast to conventional monospecific antibodies that do not have this capability. Thus, the bispecific antibodies of the invention bind two different antigens simultaneously and/or sequentially.

The "affinity", expressed by the equilibrium constant (KD) for the dissociation of an antigen from an antigen-binding polypeptide, is a measure of the strength of binding between an epitope and an antigen-binding site on an antibody (or antigen-binding fragment thereof). The smaller the KD value, the stronger the binding strength between the epitope and the antigen-binding polypeptide. Alternatively, affinity can be expressed as affinity constant (KA), i.e., 1/KD. Affinity can be determined by known methods based on the particular antigen of interest. For example, KD can be determined by surface plasmon resonance.

Any KD value less than 10 ^-6 is considered to indicate binding. Specific binding of an antibody or antigen binding fragment thereof to an antigen or antigenic determinant may be determined in any suitable known manner, including, for example, scatchard analysis (SCATCHARD ANALYSIS) and/or competitive binding assays, such as Radioimmunoassays (RIA), enzyme Immunoassays (EIA) and sandwich competition assays, equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance or spectrometry (e.g., using fluorescent assays), as well as different variants thereof known in the art.

"Avidity" is a measure of the strength of binding between an antibody or antigen-binding fragment thereof and the antigen of interest. Avidity is related to the affinity between an epitope and its antigen binding site on an antibody, and the number of relevant binding sites present on the antibody.

"In situ" means in a natural or original position, rather than being moved to another position. For example, in situ vδ1+ cells of a patient refer to vδ1 cells in vivo, as opposed to in vitro or ex vivo cells.

"Human tissue vδ1+ cells" and "hematopoietic and blood vδ1+ cells" and "Tumor Infiltrating Lymphocytes (TIL) vδ1+ cells" are defined as vδ1+ cells contained in or derived from human tissue or hematopoietic system or human tumor, respectively. All of the cell types can be identified by (i) their position or positions from which they are derived and (ii) their vδ1+ TCR expression.

A "regulatory antibody" is an antibody that confers a measurable change upon contact or binding to a cell expressing a target to which the antibody binds, including, but not limited to, a cell cycle, and/or cell number, and/or cell viability, and/or secretion of one or more cell surface markers, and/or one or more secreted molecules (e.g., cytokines, chemokines, leukotrienes, etc.), and/or a measurable change in function (e.g., cytotoxicity to a target cell or diseased cell). A method of "modulating" a cell or population thereof refers to a method of causing at least one measurable change in the secretion of the one or more cells or thereof to produce one or more "modulated cells".

An "immune response" is a measurable change in at least one cell, one cell type, one endocrine pathway, or one exocrine pathway of the immune system (including but not limited to a cell-mediated response, a humoral response, a cytokine response, a chemokine response) following the addition of a regulatory antibody.

"Immune cells" are defined as cells of the immune system, including but not limited to CD34+ cells, B cells, CD45+ (lymphocyte common antigen) cells, alpha-beta T cells, cytotoxic T cells, helper T cells, plasma cells, neutrophils, monocytes, macrophages, erythrocytes, platelets, dendritic cells, phagocytes, granulocytes, innate lymphocytes, natural Killer (NK) cells, and γδ T cells. Typically, immune cells are classified by means of combinatorial cell surface molecular analysis (e.g., via flow cytometry) to identify or group or cluster immune cells into subpopulations. These can be further subdivided with additional analysis. For example, cd45+ lymphocytes can be further subdivided into a vδ positive population and a vδ negative population.

A "model system" is a biological model or biological representation intended to aid in understanding how a drug, such as an antibody or antigen binding fragment thereof, acts like a drug in ameliorating a sign or symptom of a disease. Such models typically involve the use of diseased cells, non-diseased cells, healthy cells, effector cells, tissues, etc., in vitro, ex vivo, and in which the performance of the drug is studied and compared.

"Diseased cells" exhibit a phenotype associated with a disease (e.g., cancer), an infection (e.g., viral infection), or the progression of an inflammatory disorder or disease. For example, the diseased cell may be a tumor cell, an autoimmune tissue cell, or a virus-infected cell. Thus, the diseased cell may be defined as a tumor cell or a virally infected cell or an inflammatory cell.

"Healthy cells" refers to normal cells without lesions. They may also be referred to as "normal" or "non-diseased" cells. Non-diseased cells include non-cancerous or non-infectious or non-inflammatory cells. The cells are typically used with relevant diseased cells to determine the diseased cell specificity conferred by the drug and/or to better understand the therapeutic index of the drug.

"Diseased cell specificity" is a measure of the effectiveness of effector cells or populations thereof (e.g., like vδ1+ cell populations) in differentiating and killing diseased cells (e.g., cancer cells) while retaining non-diseased or healthy cells. This potential can be measured in a model system and can involve comparing the propensity of an effector cell or population of effector cells to selectively kill or lyse diseased cells with the potential of the effector cells to kill or lyse non-diseased or healthy cells. The diseased cell specificity may inform the drug of the potential therapeutic index.

"Enhanced diseased cell specificity" describes a phenotype of effector cells such as, for example, vδ1+ cells or populations thereof that have been modulated to further increase their ability to specifically kill diseased cells. This enhancement can be measured in a number of ways, including a fold or percentage increase in lesion cell killing specificity or selectivity.

"ADCC" or "antibody-dependent cell-mediated cytotoxicity" describes an immune response to cells coated with antibodies that bind to cell surface antigens. This is a cell-mediated process whereby immune effector cells (such as NK cells, for example) recognize cell-bound antibodies, thereby triggering degranulation and lysis of target cells. Typically, this is mediated via Fc-fcγ interactions. The Fc region of a cell-bound antibody recruits effector cells (e.g., NK cells) expressing fcγ receptors, resulting in degranulation of effector cells and death of target cells.

"Fc effective" refers to an antibody comprising a functional Fc region (fragment crystallizable region), i.e., an Fc region that has not been disabled by mutation or other means. Fc-effective antibodies exhibit unabated Fc function. An Fc-effective antibody may comprise a human IGHC heavy chain sequence as listed in IMGT, which has not been modified or engineered or constructed to reduce binding to one or more fcγ receptors. For example, via IGHC hinge mutations or by constructing antibodies comprising heavy chain constant domains that are chimeric or hybridized to the IgG1/IgG2A or IgG1/IgG4 IGHC sequences.

Suitably, the antibodies or antigen binding fragments (i.e., polypeptides) of the invention are isolated. An "isolated" polypeptide is one that has been removed from its original environment. The term "isolated" may be used to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds vδ1 or a fragment thereof is substantially free of antibodies that specifically bind antigens other than vδ1). The term "isolated" may also be used to refer to preparations in which the isolated antibody is sufficiently pure to be administered therapeutically when formulated as an active ingredient of a pharmaceutical composition, or is at least 70-80% (w/w) pure, more preferably at least 80-90% (w/w) pure, even more preferably 90-95% pure; and, most preferably, at least 95%, 96%, 97%, 98%, 99% or 100% (w/w) pure.

Suitably, the polynucleotide used in the present invention is isolated. An "isolated" polynucleotide is a polynucleotide that has been removed from its original environment. For example, a naturally occurring polynucleotide is isolated if it is separated from some or all of the coexisting materials in the natural system. For example, a polynucleotide is considered isolated if it is cloned into a vector that is not part of its natural environment, or if it is contained in a cDNA.

An antibody or antigen binding fragment thereof may be a "functionally active variant," which also includes naturally occurring allelic variants, as well as mutants or any other non-naturally occurring variants. As known in the art, an allelic variant is an alternative form of a (poly) peptide, characterized by substitution, deletion, or addition of one or more amino acids that do not substantially alter the biological function of the polypeptide. As non-limiting examples, the functionally active variants can still function when the framework containing the CDRs is modified, when the CDRs themselves are modified, when the CDRs are grafted onto alternative frameworks, or when incorporated into N-terminal or C-terminal extensions. Further, the CDR-containing binding domain can be paired with a different partner chain (e.g., a partner chain shared with another antibody). After sharing with a so-called 'universal' light chain or 'universal' heavy chain, the binding domain can still function. Further, the binding domain may function upon multimerization. Further, 'antibodies or antigen binding fragments thereof' may also comprise functional variants in which VH or VL or constant domains have been modified away from or towards different canonical sequences (e.g. as listed in imgt. Org) and still function.

To compare two closely related polypeptide sequences, the "percent sequence identity" between a first polypeptide sequence and a second polypeptide sequence can be calculated using NCBI BLAST v2.0 using the standard set of polypeptide sequences (BLASTP). To compare two closely related polynucleotide sequences, "% sequence identity" between a first nucleotide sequence and a second nucleotide sequence can be calculated using NCBI BLAST v2.0 using the standard set-up (BLASTN) of nucleotide sequences.

A polypeptide or polynucleotide sequence is said to be identical or "identical" to another polypeptide or polynucleotide sequence if the polypeptide or polynucleotide sequence shares 100% sequence identity over their entire length. Residues in the sequence are numbered from left to right, i.e., from N-terminus to C-terminus for the polypeptide; numbering is from the 5 'to 3' end for polynucleotides.

In some embodiments, any given% sequence identity of a sequence is calculated in the absence of sequences for all 6 CDRs of an antibody. For example, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: a variable heavy chain region sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a specified variable heavy chain region sequence; and/or a variable light chain region sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the specified variable light chain region sequence, wherein any amino acid variation occurs only in the framework regions of the variable heavy and light chain region sequences. In such embodiments, an anti-vδ1 antibody or antigen-binding fragment thereof having a certain sequence identity retains the complete heavy and light chain CDR1, CDR2 and CDR3 sequences of the corresponding anti-vδ1 antibody or antigen-binding fragment thereof. In more specific examples, although in NO way limiting and merely to further illustrate these embodiments of the invention, an anti-vδ1 antibody or antigen binding fragment thereof is provided comprising a polypeptide that hybridizes to SEQ ID NO:15 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:40 or a VL consisting of said amino acid sequences, wherein any amino acid variation occurs only in the framework regions of the variable heavy and light chain region sequences. Thus, the antibody of this particular example further comprises a polypeptide comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 comprising the amino acid sequence of SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, and VLCDR3 of the amino acid sequence of seq id no.

Furthermore, the antibodies and antigen binding fragments thereof provided herein may comprise a kappa light chain variable sequence and retain a non-serine amino acid residue, e.g., a non-polar and/or non-germline residue, at position 74 according to the IMGT numbering system, e.g., they comprise a leucine residue at that position. For example, although in NO way limiting and merely to further illustrate these embodiments of the invention, an anti-vδ1 antibody or antigen binding fragment thereof is provided comprising a polypeptide that hybridizes to SEQ ID NO:15 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:40, wherein any amino acid variation occurs only in the framework regions of the variable heavy and light chain region sequences, and wherein the antibody comprises a kappa light chain variable sequence comprising an amino acid residue that is non-germline and/or non-polar at position 74 according to the IMGT numbering system (e.g., a leucine residue at that position). The antibody of this embodiment further comprises a polypeptide comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 comprising the amino acid sequence of SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, and VLCDR3 of the amino acid sequence of seq id no.

"Difference" between sequences refers to the insertion, deletion or substitution of a single amino acid residue in one position of a second sequence as compared to a first sequence. Two polypeptide sequences may contain one, two or more such amino acid differences. Insertions, deletions or substitutions in the second sequence that are otherwise identical to the first sequence (100% sequence identity) result in a decrease in% sequence identity. For example, if the same sequence is 9 amino acid residues long, one substitution in the second sequence results in 88.9% sequence identity. If the first polypeptide sequence and the second polypeptide sequence are 9 amino acid residues long and share 6 identical residues, the first polypeptide sequence and the second polypeptide sequence share greater than 66% identity (the first polypeptide sequence and the second polypeptide sequence share 66.7% identity).

Or the number of additions, substitutions and/or deletions made to the first sequence for the purpose of comparing the first reference polypeptide sequence to the second comparison polypeptide sequence may be determined. "adding" is the addition of an amino acid residue to the sequence of a first polypeptide (including at either end of the first polypeptide). "substitution" is the replacement of one amino acid residue in the sequence of a first polypeptide with a different amino acid residue. The substitutions may be conservative or non-conservative. A "deletion" is a deletion of one amino acid residue from the sequence of a first polypeptide (including a deletion at either end of the first polypeptide).

Using three-letter codes and one-letter codes, naturally occurring amino acids can be referred to as follows: glycine (G or Gly), alanine (a or Ala), valine (V or Val), leucine (L or Leu), isoleucine (I or lie), proline (P or Pro), phenylalanine (F or Phe), tyrosine (Y or Tyr), tryptophan (W or Trp), lysine (K or Lys), arginine (R or Arg), histidine (H or His), aspartic acid (D or Asp), glutamic acid (E or Glu), asparagine (N or Asn), glutamine (Q or gin), cysteine (C or Cys), methionine (M or Met), serine (S or Ser) and threonine (T or Thr). Where the residue may be aspartic acid or asparagine, the symbols Asx or B may be used. When the residue may be any amino acid, the symbols Xaa or X may be used. In case the residue may be glutamic acid or glutamine, the notation Glx or Z may be used. Unless the context indicates otherwise, reference to aspartic acid includes salts of aspartic acid, and reference to glutamic acid includes salts of glutamic acid.

As used herein, numbering of polypeptide sequences and definition of CDRs and FR are as defined according to EU and/or IMGT numbering systems, as the context indicates. The "corresponding" amino acid residues between the first polypeptide sequence and the second polypeptide sequence are amino acid residues in the affinity of the first sequence that share the same position according to the EU and/or IMGT numbering system as set forth above and below as compared to the amino acid residues in the second sequence, while the amino acid residues in the second sequence may be different from the amino acid residues in the first sequence. Suitably, if the length of the framework and CDR are the same, as defined by EU or IMGT, the corresponding residues will share the same numbers (and letters). Alignment can be achieved manually or by using known computer algorithms, e.g., for sequence alignment, e.g., using the standard set-up NCBI BLAST v2.0 (BLASTP or BLASTN).

Reference herein to an "epitope" refers to a portion of a target that is specifically bound by an antibody or antigen-binding fragment thereof. Epitopes may also be referred to as "antigenic determinants". An antibody binds a "substantially identical epitope" when both antibodies recognize the same or a spatially overlapping epitope with the other antibody. A common method of determining whether two antibodies bind to the same or overlapping epitope is a competition assay, which can be configured in a variety of different formats using labeled antigens or labeled antibodies (e.g., using radioactive or enzyme-labeled well plates, or flow cytometry on antigen-expressing cells). When one antibody recognizes the same epitope as the other (i.e., all contact points between antigen and antibody are the same), the two antibodies bind to the "same epitope".

Epitopes found on a protein target may be defined as "linear epitopes" or "conformational epitopes". Linear epitopes are formed by contiguous amino acid sequences in protein antigens. Conformational epitopes are formed by amino acids that are discontinuous in the protein sequence, but which aggregate together after the protein is folded into its three-dimensional structure.

The term "vector" as used herein is intended to mean a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian and yeast vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. In addition, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors of utility in recombinant DNA technology are often in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably as the plasmid is the most commonly used form of vector. However, the present invention is intended to include such other forms of expression vectors that provide equivalent functions, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), and also phage and phagemid systems. The term "recombinant host cell" (or simply "host cell") as used herein is intended to refer to a cell into which a recombinant expression vector has been introduced. Such terms are intended to refer not only to a particular subject cell, but also to the progeny of such a cell, for example, when the progeny are employed to prepare a cell line or cell bank, which is then optionally stored, provided, sold, transferred, or used to make an antibody or antigen-binding fragment thereof as described herein.

References to "subject," "patient," or "individual" refer to a subject, particularly a mammalian subject, to be treated. Mammalian subjects include humans, non-human primates, farm animals (e.g., cows), sport animals, or pet animals (e.g., dogs, cats, guinea pigs, rabbits, rats, or mice). In some embodiments, the subject is a human. In alternative embodiments, the subject is a non-human mammal, such as a mouse.

The term "sufficient amount" means an amount sufficient to produce the desired effect. The term "therapeutically effective amount" is an amount effective to ameliorate a symptom of a disease or disorder. A therapeutically effective amount may be a "therapeutically effective amount" because control may be considered treatment.

A disease or disorder is "improved" if the severity of the sign or symptom of the disease or disorder, the frequency with which the subject experiences such sign or symptom, or both are reduced (as compared to an earlier point in time, e.g., prior to administration of any antibody).

As used herein, "treating a disease or disorder" means reducing the frequency and/or severity of at least one sign or symptom of a disease or disorder experienced by a subject (as compared to an earlier point in time, e.g., prior to administration of any antibody).

As used herein, "cancer" refers to abnormal growth or division of cells. In general, cancer cells grow and/or longevity beyond and uncoordinated with normal cells and their surrounding tissues. Cancers may be benign, premalignant, or malignant. Cancers occur in a variety of cells and tissues, including the oral cavity (e.g., mouth, tongue, pharynx, etc.), digestive system (e.g., esophagus, stomach, small intestine, colon, rectum, liver, bile duct, gall bladder, pancreas, etc.), respiratory system (e.g., larynx, lung, bronchi, etc.), bone, joint, skin (e.g., basal cell, squamous cell, meningioma, etc.), breast, reproductive system (e.g., uterus, ovary, prostate, testis, etc.), urinary system (e.g., bladder, kidney, ureter, etc.), eye, nervous system (e.g., brain, etc.), endocrine system (e.g., thyroid, etc.), and hematopoietic system (e.g., lymphoma, myeloma, leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, etc.).

As used herein, the term "about" as used herein includes up to 10% and including 10% and up to 10% and including 10% higher than the specified value, suitably up to 5% and including 5% and up to 5% and including 5% lower than the specified value, particularly the specified value. The term "between … …" includes values specifying boundaries.

Antibodies and antigen binding fragments thereof

Provided herein are antibodies or antigen binding fragments thereof capable of specifically binding to the delta variable 1 chain (vδ1) of a γδ T Cell Receptor (TCR). The present invention relates to the use of said antibodies as a medicament for administration to a subject to be treated.

In one embodiment, the antibody or antigen-binding fragment thereof is scFv, fab, fab ', F (ab') 2, fv, variable domain (e.g., VH or VL), diabody, minibody, or monoclonal antibody. In another embodiment, the antibody or antigen binding fragment thereof is an scFv.

The antibodies of the invention may be of any class, e.g., igG, igA, igM, igE, igD or an isoform thereof, and may comprise kappa or lambda light chains. In one embodiment, the antibody is an IgG antibody, e.g., at least one of the isotypes IgG1, igG2, igG3, or IgG 4. In another embodiment, the antibody may be in a format, such as an IgG format, that has been modified to confer a desired property (e.g., fc is mutated to reduce effector function, extend half-life, alter ADCC, or improve hinge stability). Such modifications are well known in the art.

In one embodiment, the antibody or antigen binding fragment thereof is human. Thus, an antibody or antigen binding fragment thereof may be derived from a human immunoglobulin (Ig) sequence. The CDRs, framework and/or constant regions of the antibody (or antigen binding fragment thereof) can be derived from human Ig sequences, in particular human IgG sequences. For human Ig sequences, particularly human IgG sequences, the CDRs, framework and/or constant regions can be substantially identical. The advantage of using human antibodies is that they have low or no immunogenicity in humans.

The antibody or antigen binding fragment thereof may also be chimeric, e.g., a mouse-human antibody chimeric.

Or the antibody or antigen binding fragment thereof is derived from a non-human species, such as a mouse. Such non-human antibodies may be modified to increase their similarity to antibody variants naturally occurring in humans, and thus the antibodies or antigen binding fragments thereof may be partially or fully humanized. Thus, in one embodiment, the antibody or antigen binding fragment thereof is humanized.

Summary of specific antibodies provided herein

Summary of some specific antigen binding molecules (i.e., antibodies) provided by the present invention, as well as the identification of the assigned SEQ ID NOs in the accompanying sequence listing, are provided below. Antigen binding variants, derivatives and fragments thereof are also provided as part of the invention. The sequences are provided in the attached sequence listing and figures. If there is any difference between the sequences in the sequence table and the sequences in FIGS. 45 to 48, the sequences in the figures should be taken into account.

TABLE 1 summary of antibodies (heavy chain) derived from ADT1-4 and related SEQ ID NOs

Antibodies to	VH	HFR1	VHCDR1	HFR2	VHCDR2	HFR3	VHCDR3	HFR4
									Parental ADT1-4	1	170	51	172	53	173	54	174
ADT1-4-105	2	170	51	172	53	173	55	174
									ADT1-4-107	3	170	51	172	53	173	56	174
ADT1-4-110	4	170	51	172	53	173	57	174
									ADT1-4-112	5	170	51	172	53	173	58	174
ADT1-4-117	6	170	51	172	53	173	59	174
									ADT1-4-19	7	170	51	172	53	173	60	174
ADT1-4-21	8	170	52	172	53	173	61	174
									ADT1-4-31	9	170	51	172	53	173	62	174
ADT1-4-139	10	170	51	172	53	173	63	174
									ADT1-4-4	11	170	51	172	53	173	64	174
ADT1-4-143	12	171	51	172	53	173	65	174
									ADT1-4-53	13	170	52	172	53	173	66	174
ADT1-4-173	14	170	51	172	53	173	67	174
									ADT1-4-2	15	170	51	172	53	173	68	174
ADT1-4-8	16	170	51	172	53	173	69	174
									ADT1-4-82	17	170	51	172	53	173	70	174
ADT1-4-83	18	170	51	172	53	173	71	174
									ADT1-4-3	19	170	51	172	53	173	72	174
ADT1-4-84	20	170	51	172	53	173	73	174
									ADT1-4-86	21	170	52	172	53	173	74	174
ADT1-4-95	22	170	51	172	53	173	75	174
									ADT1-4-1	23	170	51	172	53	173	76	174
ADT1-4-6	24	170	51	172	53	173	77	174
									ADT1-4-138	25	170	51	172	53	173	78	174

TABLE 2 summary of antibodies derived from ADT1-4 (light chain) and related SEQ ID NOs

TABLE 3 summary of antibodies (heavy chain) derived from ADT1-7 and related SEQ ID NOs

Antibodies to	VH	HFR1	VHCDR1	HFR2	VHCDR2	HFR3	VHCDR3	HFR4
									Parental ADT1-4	106	189	130	190	131	191	132	192
ADT1-7-10	107	189	130	190	131	191	133	192
									ADT1-7-15	108	189	130	190	131	191	134	192
ADT1-7-17	109	189	130	190	131	191	135	192
									ADT1-7-18	110	189	130	190	131	191	136	192
ADT1-7-19	111	189	130	190	131	191	137	192
									ADT1-7-20	112	189	130	190	131	191	138	192
ADT1-7-22	113	189	130	190	131	191	139	192
									ADT1-7-23	114	189	130	190	131	191	140	192
ADT1-7-42	115	189	130	190	131	191	141	192
									ADT1-7-3	116	189	130	190	131	191	142	192
ADT1-7-61	117	189	130	190	131	191	143	192

TABLE 4 summary of antibodies derived from ADT1-7 (light chain) and related SEQ ID NOs

TABLE 5 summary of other parent antibodies and related SEQ ID NOs

Parent antibodies	VH	VL
			C08	273	282
B07	274	283
			C05	275	284
E04	276	285
			F07	277	286
G06	278	287
			G09	279	288
B09	280	289
			G10	281	290
E01	312	313

ADT 1-4-and ADT 1-7-derived antibodies

The present invention provides antibodies derived from parent antibody ADT1-4 (having a variable heavy chain region sequence according to SEQ ID NO:1 and a variable light chain region sequence according to SEQ ID NO: 26), and antibodies derived from parent antibody ADT1-7 (having a variable heavy chain region sequence according to SEQ ID NO:106 and a variable light chain region sequence according to SEQ ID NO: 118). ADT1-4 is also referred to herein as G04, and ADT1-4 and G04 are used interchangeably. ADT1-7 is also referred to herein as E07, and ADT1-7 and E07 are used interchangeably.

In some embodiments, the invention provides an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 55 to 78 and 133 to 143, a VHCDR3 having or consisting of an amino acid sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the sequence of the group consisting of VHCDR 3; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 82 to 105 and 147 to 157 has an amino acid sequence of or a VLCDR3 consisting of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In some embodiments, the invention provides an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, the VHCDR1 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO: 51. 52 and 130;

A VHCDR2, the VHCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:53 and 131; and

A VHCDR3 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:55 to 78 and 133 to 143; and

A light chain variable region comprising:

A VLCDR1, the VLCDR1 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:79 and 144;

A VLCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:80 and 145; and

A VLCDR3 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:82 to 105 and 147 to 157.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In some embodiments, the invention provides an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 2 to 25 and 107 to 117 has or consists of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 27 to 50 and 119 to 129 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

ADT1-4 derived antibodies

The present invention provides antibodies derived from parent antibody ADT1-4 (having a variable heavy chain region sequence according to SEQ ID NO:1 and a variable light chain region sequence according to SEQ ID NO: 26), e.g., as set forth below. ADT1-4 is also referred to herein as G04, and ADT1-4 and G04 are used interchangeably.

Antibodies comprising specific CDR sequences derived from ADT1-4

Antibodies provided herein include the following antibodies having specific sequences derived from ADT 1-4.

For example, in some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody, or antigen-binding fragment thereof, comprising: a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 55 to 78 or a VHCDR3 consisting of said amino acid sequences; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 82 to 105 or a VLCDR3 consisting of said amino acid sequences. Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 55 to 77 or a VHCDR3 consisting of said amino acid sequences; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 82 to 104 or a VLCDR3 consisting of said amino acid sequence. Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 58. 60, 61, 62, 65, 66, 68, 74, 76 and 77 or a VHCDR3 consisting of said amino acid sequences; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 85. 87, 88, 89, 92, 93, 95, 101, 103 and 104 or a VLCDR3 consisting of said amino acid sequences. Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, said VHCDR1 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:51 and 52;

A VHCDR2, said VHCDR2 comprising SEQ ID NO:53 or consists of said amino acid sequence; and

A VHCDR3, said VHCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:55 to 78; and

A light chain variable region comprising:

A VLCDR1, said VLCDR1 comprising the amino acid sequence of SEQ ID NO:79 or consists of the amino acid sequence of seq id no;

a VLCDR2, said VLCDR2 comprising SEQ ID NO:80 or consists of the amino acid sequence of seq id no; and

A VLCDR3, the VLCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:82 to 105.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, said VHCDR1 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:51 and 52;

A VHCDR2, said VHCDR2 comprising SEQ ID NO:53 or consists of said amino acid sequence; and

A VHCDR3, said VHCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:55 to 77; and

A light chain variable region comprising:

A VLCDR1, said VLCDR1 comprising the amino acid sequence of SEQ ID NO:79 or consists of the amino acid sequence of seq id no;

a VLCDR2, said VLCDR2 comprising SEQ ID NO:80 or consists of the amino acid sequence of seq id no; and

A VLCDR3, the VLCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:82 to 104.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, said VHCDR1 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO:51 and 52;

A VHCDR2, said VHCDR2 comprising SEQ ID NO:53 or consists of said amino acid sequence; and

A VHCDR3, said VHCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO: 58. 60, 61, 62, 65, 66, 68, 74, 76 and 77; and

A light chain variable region comprising:

A VLCDR1, said VLCDR1 comprising the amino acid sequence of SEQ ID NO:79 or consists of the amino acid sequence of seq id no;

a VLCDR2, said VLCDR2 comprising SEQ ID NO:80 or consists of the amino acid sequence of seq id no; and

A VLCDR3, the VLCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NO: 85. 87, 88, 89, 92, 93, 95, 101, 103 and 104.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-V delta 1) antibody or antigen binding fragment thereof comprising

A) Comprising SEQ ID NOs: 51. 53 and 55 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 82 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

b) Comprising SEQ ID NOs: 51. 53 and 56 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 83 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

c) Comprising SEQ ID NOs: 51. 53 and 57 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 84, or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

d) Comprising SEQ ID NOs: 51. 53 and 58 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 85 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

e) Comprising SEQ ID NOs: 51. 53 and 59 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 86 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

f) Comprising SEQ ID NOs: 51. 53 and 60 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 87 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

g) Comprising SEQ ID NOs: 52. 53 and 61 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 88 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

h) Comprising SEQ ID NOs: 51. 53 and 62 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 89 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

i) Comprising SEQ ID NOs: 51. 53 and 63 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 90 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

j) Comprising SEQ ID NOs: 51. 53 and 64 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 91 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

k) Comprising SEQ ID NOs: 51. 53 and 65 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 92 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

l) each comprises the sequence of SEQ ID NO: 52. 53 and 66 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 93, or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

m) each comprises the sequence of SEQ ID NO: 51. 53 and 67 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 94 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

n) each comprises the sequence of SEQ ID NO: 51. 53 and 68 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 95 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

o) comprises the amino acid sequences of SEQ ID NOs: 51. 53 and 69 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 96 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

p) each comprises the sequence of SEQ ID NO: 51. 53 and 70 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 97 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

q) each comprises SEQ ID NO: 51. 53 and 71 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 98 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

r) each comprises the sequence of SEQ ID NO: 51. 53 and 72 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 99 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

s) comprise the amino acid sequences of SEQ ID NOs: 51. 53 and 73 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 100 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

t) comprises the amino acid sequence of SEQ ID NO: 52. 53 and 74 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 101 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

u) each comprises SEQ ID NO: 51. 53 and 75 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 102 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

v) comprises the amino acid sequences of SEQ ID NOs: 51. 53 and 76 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 103 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

w) comprises the amino acid sequence of SEQ ID NO: 51. 53 and 77 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 104 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences; or;

x) comprises the amino acid sequence of SEQ ID NO: 51. 53 and 78 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 105 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences.

The anti-vδ1 antibody or antigen binding fragment thereof may comprise a kappa light chain variable sequence (or comprise a variable light chain derived from a kappa light chain variable sequence) wherein the residue at position 74 of the kappa light chain variable sequence is not serine, e.g. a non-human germline residue at position 74 and/or a non-polar residue, e.g. the residue at position 74 is a leucine residue, according to the IMGT numbering system.

Other embodiments are provided below.

ADT1-4-105 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-105 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:55, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. And/or a light chain variable region comprising a sequence identical to SEQ ID NO:82 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, there is provided an antibody or antigen binding fragment or variant thereof j comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:55, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:82 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:55, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:82, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-105, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:55, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:82, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:55, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:82, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:55, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:82, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:55, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:82, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-105, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:55, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:82, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-107 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-107 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:56 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:83 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:56, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:83, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:56, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:83, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-107, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:56, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:83, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:56, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:83, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:56, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:83, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:56, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:83, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-107, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:56, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:83, optionally comprising 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-110 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-110 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:57, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:84 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:57, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:84, the amino acid sequence has a VLCDR3 of an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:57, a VHCDR3 of an amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:84, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-110, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:57, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:84, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:57, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:84, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:57, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:84, and a VLCDR3 of at least 90% identity to the amino acid sequence of 84.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:57, a VHCDR3 of an amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:84, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-110, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:57, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:84, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-112 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-112 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:58, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. And/or a light chain variable region comprising a sequence identical to SEQ ID NO:85, has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, there is provided an antibody or antigen binding fragment or variant thereof j comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:58, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:85, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:58, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:85, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-112, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:58, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:85, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:58, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:85, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:58, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:85, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:58, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:85, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-112, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:58, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:85, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-117 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-117 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:59 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:86 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:59, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:86 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:59, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:86, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-117, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:59, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:86, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:59, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:86, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:59, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:86, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:59, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:86, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-117, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:59, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:86, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-19 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-19 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:60, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of the VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:87 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:60, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of 60; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:87 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:60, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:87, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-19, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:60, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:87, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:60, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:87, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:60, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:87, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:60, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:87, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-19, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:60, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:87, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-21 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-21 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:61, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:88 has an amino acid sequence of VLCDR3 of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:61, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:88 has an amino acid sequence having at least 90% identity to the VLCDR3 of the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:61, a VHCDR3 of an amino acid sequence; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:88, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-21, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:61, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:88, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:61, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:88, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:61, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:88, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:61, a VHCDR3 of an amino acid sequence; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:88, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-21, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:61, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:88, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-31 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-31 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:62, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. And/or a light chain variable region comprising a sequence identical to SEQ ID NO:89 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:62, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:89, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:62, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:89, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-31, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:62, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:89, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:5l, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:62, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:89, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:62, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:89, and a VLCDR3 of at least 90% identity to the amino acid sequence of 89.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:62, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:89, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-31, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:62, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:89, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-139 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-139 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:63, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:90 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:63, a VHCDR3 of an amino acid sequence having at least 90% identity to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:90, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:63, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:90, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-139, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:63, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:90, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:63, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:90, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:63, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:90, and a VLCDR3 of at least 90% identity to the amino acid sequence of 90.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:63, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:90, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-139, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:63, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:90, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-4 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-4 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:64, a VHCDR3 having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:91 has a VLCDR3 of an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:64, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of 64; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:91 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:64, a VHCDR3 of an amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:91, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-4, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:64, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:91, optionally comprising 1 or 2 amino acid substitutions of VLCDR3. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:64, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:91, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:64, a VHCDR3 of at least 90% identity to the amino acid sequence of 64; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:91, and a VLCDR3 of at least 90% identity to the amino acid sequence of 91.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:64, a VHCDR3 of an amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:91, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-4, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:64, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:91, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-143 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-143 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:65 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:92 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, there is provided an antibody or antigen binding fragment or variant thereof j comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:65, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:92 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:65, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:92, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-143, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:65, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:92, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:65, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:92, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:65, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:92, and a VLCDR3 of at least 90% identity to the amino acid sequence of 92.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:65, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:92, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-143, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:65, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:92, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-53 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-53 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:66, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:93 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:66, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of 66; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:93 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:66, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:93, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-53, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:66, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:93, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:66, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:93, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:66, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:93, and a VLCDR3 of at least 90% identity to the amino acid sequence of 93.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:66, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:93, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-53, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:66, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:93, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-173 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-173 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:67, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:94 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the VLCDR3 of the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:67, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:94, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:67, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:94, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-173, for example, to provide an antibody or antigen-binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:67, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:94, optionally comprising 1 or 2 amino acid substitutions of VLCDR3. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:67, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:94, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:67, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:94, and a VLCDR3 of at least 90% identity to the amino acid sequence of 94.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:67, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:94, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-173, for example, to provide an antibody or antigen-binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:67, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:94, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-2 and fragments and variants thereof

Certain embodiments relate to antibody ADT1-4-2 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:68 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:95 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the VLCDR3 of the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:68, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:95 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:95, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-2, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:68, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:95, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:68, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:95, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:68, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:95, and a VLCDR3 of at least 90% identity to the amino acid sequence of 95.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-2, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:95, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-8 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-8 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:69 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:96 has an amino acid sequence of VLCDR3 of an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:69, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:96 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:69, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:96, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-8, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:69, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:96, optionally comprising 1 or 2 amino acid substitutions of VLCDR3. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:69, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:96, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:69, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:96, and a VLCDR3 of at least 90% identity to the amino acid sequence of 96.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:69, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:96, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-8, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:69, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:96, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-82 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-82 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:70, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of the VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:97 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:70, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:97, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:70, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:97, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-82, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:70, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:97, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:70, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to a VHCDR3 of the amino acid sequence of 70; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:97, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:70, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:97, and a VLCDR3 of at least 90% identity to the amino acid sequence of 97.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID N0:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:70, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:97, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-82, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:70, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:97, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-83 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-83 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:71, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:98 has an amino acid sequence of VLCDR3 of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:71, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:98, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:71, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:98, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-83, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:71, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:98, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:71, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:98, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:71, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:98, and a VLCDR3 of at least 90% identity to the amino acid sequence of 98.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:71, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:98, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-83, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:71, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:98, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-3 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-3 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:72 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:99 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the VLCDR3 of the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:72, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:99 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:72, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:99, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-3, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:72, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:99, optionally comprising 1 or 2 amino acid substitutions of VLCDR3. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:72, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:99, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:72, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:99, and a VLCDR3 of at least 90% identity to the amino acid sequence of 99.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:72, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:99, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-3, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:72, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:99, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-84 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-84 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:73 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:100 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:73, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:100, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:73, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:100, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-84, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:73, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:100, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:73, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:100, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:73, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:100, and a VLCDR3 of at least 90% identity to the amino acid sequence of 100.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:73, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:100, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-84, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:73, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:100, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-86 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-86 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:74, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:101 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:74, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:101 has an amino acid sequence having at least 90% identity to the amino acid sequence of VLCDR3. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:74, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:101, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-86, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:74, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:101, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:74, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:101, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:52, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:74, a VHCDR3 of at least 90% identity to the amino acid sequence of 74; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:101, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:74, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:101, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-86, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:52, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:74, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:101, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-95 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-95 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:75, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. And/or a light chain variable region comprising a sequence identical to SEQ ID NO:102 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:75, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:102 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:75, a VHCDR3 of an amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:102, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-95, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:75, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:102, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:75, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:102, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:75, a VHCDR3 of at least 90% identity to an amino acid sequence of 75; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:102, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:75, a VHCDR3 of an amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:102, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-95, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:75, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:102, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-1 and fragments and variants thereof

Certain embodiments relate to antibody ADT1-4-1 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:76, at least one of the amino acid sequences of which has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:103 has an amino acid sequence of VLCDR3 of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:76, a VHCDR3 having an amino acid sequence that is at least 90% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:103 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:76, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:103, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-1, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:76, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:103, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:76, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:103, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:76, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:103, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:76, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:103, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-1, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:76, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:103, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-6 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-6 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:77, a VHCDR3 having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:104 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:77, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:104, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:77, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:104, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-6, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:77, optionally comprising a VHCDR3 of 1 or2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:104, optionally comprising a VLCDR3 of 1 or2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:77, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:104, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:77, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:104, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:77, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:104, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-6, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:77, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:104, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-4-138 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-4-138 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:78 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:105 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:78, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:105 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:78, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:105, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-138, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:78, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:105, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:53, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:78, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to SEQ ID NO:80, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:105, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:51, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:78, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:79, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:105, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:78, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:105, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-4-138, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:53, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions and a polypeptide comprising the amino acid sequence of SEQ ID NO:78, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:80, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:105, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

Antibodies comprising heavy and/or light chain variable regions derived from ADT1-4

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 25, a sequence having or consisting of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 50, has or consists of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity.

Also provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 25, wherein the heavy chain variable region sequence is not SEQ ID NO:1, a step of; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 50, wherein the light chain variable region sequence is not SEQ ID NO:26.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 24, a sequence having or consisting of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 49, has or consists of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 5,7,8,9, 12, 13, 15, 21, 23 and 24 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 30. 32, 33, 34, 37, 38, 40, 46, 48 and 49 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 25, or consists of a sequence having at least 90% identity to the sequence of the group consisting of seq id no; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 50, or a sequence having or consisting of at least 90% identity to a sequence of the group consisting of seq id no.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 24, or consists of a sequence having at least 90% identity to the sequence of the group consisting of seq id no; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 49, or a sequence having or consisting of at least 90% identity to a sequence of the group consisting of seq id no.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 5. 7, 8, 9, 12, 13, 15, 21, 23 and 24, or a sequence having at least 90% identity thereto; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 30. 32, 33, 34, 37, 38, 40, 46, 48 and 49, has or consists of a sequence having at least 90% identity.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 25 or consists of said sequence; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 50 or a sequence consisting of said sequence.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 2 to 24 or a sequence consisting of said sequence; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 27 to 49 or consists of said sequence.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 5. 7, 8, 9, 12, 13, 15, 21, 23 and 24 or a sequence consisting of said sequences; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 30. 32, 33, 34, 37, 38, 40, 46, 48 and 49 or a combination thereof.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising a sequence identical to SEQ ID NO:2 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:27 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

b) Comprising a sequence identical to SEQ ID NO:3 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:28 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

c) Comprising a sequence identical to SEQ ID NO:4 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:29 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

d) Comprising a sequence identical to SEQ ID NO:5 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:30 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

e) Comprising a sequence identical to SEQ ID NO:6 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:31 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

f) Comprising a sequence identical to SEQ ID NO:7 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:32 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

g) Comprising a sequence identical to SEQ ID NO:8 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:33 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

h) Comprising a sequence identical to SEQ ID NO:9 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:34 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

i) Comprising a sequence identical to SEQ ID NO:10 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:35 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

j) Comprising a sequence identical to SEQ ID NO:11 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:36 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

k) Comprising a sequence identical to SEQ ID NO:12 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:37 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

l) comprises a sequence identical to SEQ ID NO:13 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID N0:38 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

m) comprises a sequence identical to SEQ ID NO:14 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:39 or a VL consisting of an amino acid sequence at least 90% identical to the amino acid sequence;

n) comprises a sequence identical to SEQ ID NO:15 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:40 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

o) comprises a sequence identical to SEQ ID NO:16 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:41 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

p) comprises a sequence identical to SEQ ID NO:17 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:42 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

q) comprises a sequence identical to SEQ ID NO:18 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:43 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

r) comprises a sequence identical to SEQ ID NO:19 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:44 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

s) comprises a sequence identical to SEQ ID NO:20 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:45 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

t) comprises a sequence identical to SEQ ID NO:21 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:46 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

u) comprises a sequence identical to SEQ ID NO:22 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:47 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

v) comprises a sequence identical to SEQ ID NO:23 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:48 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

w) comprises a sequence identical to SEQ ID NO:24 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID N0:49 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence; or (b)

X) comprises a sequence identical to SEQ ID NO:25 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:50 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising a sequence identical to SEQ ID NO:2 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:27 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

b) Comprising a sequence identical to SEQ ID NO:3 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:28 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

c) Comprising a sequence identical to SEQ ID NO:4 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:29 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

d) Comprising a sequence identical to SEQ ID NO:5 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:30 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

e) Comprising a sequence identical to SEQ ID NO:6 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:31 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

f) Comprising a sequence identical to SEQ ID NO:7 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:32 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

g) Comprising a sequence identical to SEQ ID NO:8 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:33 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

h) Comprising a sequence identical to SEQ ID NO:9 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:34 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

i) Comprising a sequence identical to SEQ ID NO:10 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:35 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

j) Comprising a sequence identical to SEQ ID NO:11 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:36 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

k) Comprising a sequence identical to SEQ ID NO:12 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID N0:37 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

l) comprises a sequence identical to SEQ ID NO:13 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:38 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

m) comprises a sequence identical to SEQ ID NO:14 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:39 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence;

n) comprises a sequence identical to SEQ ID NO:15 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:40 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

o) comprises a sequence identical to SEQ ID NO:16 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:41 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

p) comprises a sequence identical to SEQ ID NO:17 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:42 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence;

q) comprises a sequence identical to SEQ ID NO:18 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:43 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence;

r) comprises a sequence identical to SEQ ID NO:19 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:44 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

s) comprises a sequence identical to SEQ ID NO:20 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:45 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

t) comprises a sequence identical to SEQ ID NO:21 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:46 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

u) comprises a sequence identical to SEQ ID NO:22 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:47 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

v) comprises a sequence identical to SEQ ID NO:23 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:48 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence of seq id no;

w) comprises a sequence identical to SEQ ID NO:24 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:49 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence; or (b)

X) comprises a sequence identical to SEQ ID NO:25 or a VH consisting of or comprising an amino acid sequence at least 96% identical to the amino acid sequence of SEQ ID NO:50 or a VL consisting of an amino acid sequence that is at least 96% identical to the amino acid sequence.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising SEQ ID NO:2 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:27 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

b) Comprising SEQ ID NO:3 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:28 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

c) Comprising SEQ ID NO:4 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:29 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

d) Comprising SEQ ID NO:5 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:30 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

e) Comprising SEQ ID NO:6 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:31 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

f) Comprising SEQ ID NO:7 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:32 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

g) Comprising SEQ ID NO:8 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:33 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

h) Comprising SEQ ID NO:9 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:34 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

i) Comprising SEQ ID NO:10 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:35 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

j) Comprising SEQ ID NO:11 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:36 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

k) Comprising SEQ ID NO:12 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:37 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

l) a polypeptide comprising SEQ ID NO:13 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:38 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

m) comprises the sequence of SEQ ID NO:14 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:39 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

n) comprises the sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

o) comprises SEQ ID NO:16 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:41 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

p) comprises the sequence of SEQ ID NO:17 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:42 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

q) comprises SEQ ID NO:18 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:43 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

r) comprises the sequence of SEQ ID NO:19 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:44 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

s) comprises the sequence of SEQ ID NO:20 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:45 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

t) comprises the sequence of SEQ ID NO:21 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:46 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

u) comprises SEQ ID NO:22 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:47 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

v) a polypeptide comprising SEQ ID NO:23 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:48 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

w) comprises SEQ ID NO:24 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:49 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions; or (b)

X) comprises SEQ ID NO:25 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:50 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions,

Optionally wherein the VH sequence is not SEQ ID NO:1 and/or VL sequence other than SEQ ID NO:26.

"Spans two variable regions" collectively refers to that an antibody may contain up to a specified number of substitutions when considering both heavy and light chain variable regions. Amino acid substitutions may be conservative amino acid substitutions. In some embodiments, substitution (if present) may occur anywhere in the variable region sequence. In preferred embodiments, substitution (if any) may be limited to the framework regions. Thus, in some embodiments, amino acid substitutions do not occur in the CDR sequences.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising SEQ ID NO:2 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:27 or a VL consisting of said amino acid sequence;

b) Comprising SEQ ID NO:3 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:28 or a VL consisting of said amino acid sequence;

c) Comprising SEQ ID NO:4 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:29 or a VL consisting of said amino acid sequence;

d) Comprising SEQ ID NO:5 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:30 or a VL consisting of said amino acid sequence;

e) Comprising SEQ ID NO:6 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:31 or a VL consisting of said amino acid sequence;

f) Comprising SEQ ID NO:7 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:32 or a VL consisting of said amino acid sequence;

g) Comprising SEQ ID NO:8 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:33 or a VL consisting of said amino acid sequence;

h) Comprising SEQ ID NO:9 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:34 or a VL consisting of said amino acid sequence;

i) Comprising SEQ ID NO:10 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:35 or a VL consisting of said amino acid sequence;

j) Comprising SEQ ID NO:11 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:36 or a VL consisting of said amino acid sequence;

k) Comprising SEQ ID NO:12 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:37 or a VL consisting of said amino acid sequence;

l) a polypeptide comprising SEQ ID NO:13 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:38 or a VL consisting of said amino acid sequence;

m) comprises the sequence of SEQ ID NO:14 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:39 or a VL consisting of said amino acid sequence;

n) comprises the sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence;

o) comprises SEQ ID NO:16 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:41 or a VL consisting of said amino acid sequence;

p) comprises the sequence of SEQ ID NO:17 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:42 or a VL consisting of said amino acid sequence;

q) comprises SEQ ID NO:18 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:43 or a VL consisting of said amino acid sequence;

r) comprises the sequence of SEQ ID NO:19 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:44 or a VL consisting of said amino acid sequence;

s) comprises the sequence of SEQ ID NO:20 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:45 or a VL consisting of said amino acid sequence;

t) comprises the sequence of SEQ ID NO:21 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:46 or a VL consisting of said amino acid sequence;

u) comprises SEQ ID NO:22 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:47 or a VL consisting of said amino acid sequence;

v) a polypeptide comprising SEQ ID NO:23 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:48 or a VL consisting of said amino acid sequence;

w) comprises SEQ ID NO:24 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:49 or a VL consisting of said amino acid sequence; or (b)

X) comprises SEQ ID NO:25 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:50 or a VL consisting of said amino acid sequence.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In any embodiment involving a defined VH and/or VL sequence (e.g., defined as any VH and/or VL sequence having a certain percentage of identity and/or substitution), preferably the VH sequence is not SEQ ID NO:1 and the VL sequence is not SEQ ID NO:26.

Other antibodies derived from ADT1-4

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising GDSVSSKSX ₁ a (SEQ ID NO:158 Or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:53 or consists of said sequence

C) An HCDR3 sequence comprising X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO:162 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:54;

d) An LCDR1 sequence comprising SEQ ID NO:79 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:80 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO:166 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:81;

And wherein each of X1 to X13 is a naturally occurring amino acid.

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising GDSVSSKSX ₁ a (SEQ ID NO:159 Or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:53 or consists of said sequence

C) An HCDR3 sequence comprising X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO:163 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:54;

d) An LCDR1 sequence comprising SEQ ID NO:79 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:80 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO:167 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:81;

and wherein: x ₁ is selected from the group consisting of: a and V; x ₂ is selected from the group consisting of: s and T; x ₃ is selected from the group consisting of: v, A and L; x ₄ is selected from the group consisting of: G. e and D; x ₅ is selected from the group consisting of: y and N; x ₆ is selected from the group consisting of: v, A and P; x ₇ is selected from the group consisting of: v, Y and R; x ₈ is selected from the group consisting of: K. r and G; x ₉ is selected from the group consisting of: s and K; x ₁₀ is selected from the group consisting of: t, Q, A, E and D; x ₁₁ is selected from the group consisting of: p, H and D; x ₁₂ is selected from the group consisting of: q, R, K, W, P, E and I; and X ₁₃ is selected from the group consisting of: I. v and L. The antibody may further comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO: 172; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173, a sequence of seq id no; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174, a sequence of seq id no; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO: 175; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO: 176; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or 178; and an LFR4 sequence, the LFR4 sequence comprising the sequence of SEQ ID NO: 179. 180, 181 or 182.

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising GDSVSSKSX ₁ a (SEQ ID NO:160 Or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:53 or consists of said sequence

C) An HCDR3 sequence comprising X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO:164 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:54;

d) An LCDR1 sequence comprising SEQ ID NO:79 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:80 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO:168 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:81;

And wherein X ₁ is selected from the group consisting of: a and V; x ₂ is selected from the group consisting of: s and T; x ₃ is selected from the group consisting of: v, A and L; x4 is selected from the group consisting of: G. e and D; x ₅ is selected from the group consisting of: y and N; x ₆ is selected from the group consisting of: v, A and P; x ₇ is selected from the group consisting of: v, Y and R; x ₈ is selected from the group consisting of: K. r and G; x ₉ is selected from the group consisting of: s and K; x ₁₀ is selected from the group consisting of: t, Q, A, E and D; x ₁₁ is selected from the group consisting of: p and H; x ₁₂ is selected from the group consisting of: q, R, K, W, P, E and I; and X ₁₃ is selected from the group consisting of: I. v and L. The antibody may further comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO: 172; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173, a sequence of seq id no; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174, a sequence of seq id no; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO: 175; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO: 176; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177, a sequence of seq id no; and an LFR4 sequence, the LFR4 sequence comprising SEQ ID NO: 179. 180, 181 or 182.

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising GDSVSSKSX ₁ a (SEQ ID NO:161 Or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:53 or consists of said sequence

C) An HCDR3 sequence comprising X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO:165 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:54;

d) An LCDR1 sequence comprising SEQ ID NO:79 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:80 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO:169 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:81;

And wherein X ₁ is selected from the group consisting of: a and V; x ₂ is selected from the group consisting of: s and T; x ₃ is selected from the group consisting of: v, A and L; x ₄ is selected from the group consisting of: g and D; x ₅ is selected from the group consisting of: y and N; x ₆ is selected from the group consisting of: v, A and P; x ₇ is selected from the group consisting of: v, Y and R; x ₈ is selected from the group consisting of: k and R; x ₉ is selected from the group consisting of: s and K; x ₁₀ is selected from the group consisting of: t, Q, A and E; x ₁₁ is selected from the group consisting of: p and H; x ₁₂ is selected from the group consisting of: q, K, W, P and I; and X ₁₃ is selected from the group consisting of: v and L. The antibody may further comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO: 172; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173, a sequence of seq id no; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174, a sequence of seq id no; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO: 175; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO: 176; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177, a sequence of seq id no; and an LFR4 sequence, the LFR4 sequence comprising the sequence SEQ ID NO:179 or 181.

ADT1-7 derived antibodies

The present invention provides antibodies derived from parent antibody ADT1-7 (having a variable heavy chain region sequence according to SEQ ID NO:106 and a variable light chain region sequence according to SEQ ID NO: 118), e.g., as set forth below. ADT1-7 is also referred to herein as E07, and ADT1-7 and E07 are used interchangeably.

Antibodies comprising specific CDR sequences derived from ADT1-7

Antibodies provided herein include the following antibodies having specific sequences derived from ADT 1-7.

For example, in some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody, or antigen-binding fragment thereof, comprising: a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 133 to 143 or a VHCDR3 consisting of said amino acid sequences; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID N0:147 to 157 or a VLCDR3 consisting of said amino acid sequences. Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 138. 142 and 143 or a VHCDR3 consisting of said amino acid sequences; and/or a light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 152. 156 and 157 or a VLCDR3 consisting of said amino acid sequences, certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, said VHCDR1 comprising the amino acid sequence of SEQ ID NO:130 or consists of the amino acid sequence of seq id no;

A VHCDR2, said VHCDR2 comprising SEQ ID NO:131 or consists of said amino acid sequence; and

A VHCDR3, said VHCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID No:133 to 143; and

A light chain variable region comprising:

A VLCDR1, said VLCDR1 comprising the amino acid sequence of SEQ ID NO:144 or consists of the amino acid sequence of 144;

a VLCDR2, said VLCDR2 comprising SEQ ID NO:145 or consists of said amino acid sequence; and

A VLCDR3, said VLCDR3 comprising a sequence selected from the group consisting of SEQ ID nos: 147 to 157 or consists of an amino acid sequence of the group consisting of the amino acid sequences.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, said VHCDR1 comprising the amino acid sequence of SEQ ID NO:130 or consists of the amino acid sequence of seq id no;

A VHCDR2, said VHCDR2 comprising SEQ ID NO:131 or consists of said amino acid sequence; and

A VHCDR3, said VHCDR3 comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID No:138, 142 and 143; and

A light chain variable region comprising:

A VLCDR1, said VLCDR1 comprising the amino acid sequence of SEQ ID NO:144 or consists of the amino acid sequence of 144;

a VLCDR2, said VLCDR2 comprising SEQ ID NO:145 or consists of said amino acid sequence; and

A VLCDR3, said VLCDR3 comprising a sequence selected from the group consisting of SEQ ID nos: 152. 156 and 157 or a combination of said amino acid sequences.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

Other embodiments are provided below.

ADT1-7-10 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-10 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:133, a VHCDR3 having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:147 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:133, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:147 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:133, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:147, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-10, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:133, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:147, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:133, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:147, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:133, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:147, and a VLCDR3 of at least 90% identity to the amino acid sequence of 147.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:133, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:147, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-10, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:133, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:147, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-15 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-15 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:134, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:148 has an amino acid sequence of VLCDR3 of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:134, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:148, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:134, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:148, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-15, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:134, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:148, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:134, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:148, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:134, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:148, and a VLCDR3 of at least 90% identity to the amino acid sequence of 148.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:134, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:148, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-15, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:134, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:148, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-17 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-17 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:135 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:149 has a VLCDR3 of an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:135, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:149, a VLCDR3 having an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:135, a VHCDR3 of an amino acid sequence; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:149, and a VLCDR3 of the amino acid sequence of 149.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-17, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:135, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:149, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:135, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:149, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:135, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:149, at least 90% identical VLCDR3 of the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:135, a VHCDR3 of an amino acid sequence; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:149, and a VLCDR3 of the amino acid sequence of 149.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-17, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:135, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:149, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-18 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-18 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:136 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:150 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:136, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:150 has an amino acid sequence having at least 90% identity to the amino acid sequence of VLCDR3. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:136, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:150, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-18, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:136, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:150, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:136, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:150, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:136, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:150, and a VLCDR3 of at least 90% identity to the amino acid sequence of 150.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:136, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:150, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-18, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:136, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:150, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-19 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-19 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:137, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:151 has a VLCDR3 of an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:137, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:151 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:137, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:151, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-19, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:137, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:151, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:137, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:151, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:137, at least 90% identity of the amino acid sequence of VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:151, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:137, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:151, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-19, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:137, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:151, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-20 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-20 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:138, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of the VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:152 has a VLCDR3 of an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:138, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:152 has a VLCDR3 of an amino acid sequence having at least 90% identity to the amino acid sequence of seq id no. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:138, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:152, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-20, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:138, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:152, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:138, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:152, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:138, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:152, and a VLCDR3 of at least 90% identity to the amino acid sequence of 152.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:138, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:152, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-20, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:138, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:152, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-22 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-22 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:139, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:153 have a VLCDR3 of an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:139, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of 139; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:153, and a VLCDR3 having an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO: VHCDR3 of the amino acid sequence of 139; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:153, and a VLCDR3 of the amino acid sequence of 153.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-22, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:139, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:153, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:139, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:153, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:139, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:153, and a VLCDR3 of at least 90% identity to the amino acid sequence of 153.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: VHCDR3 of the amino acid sequence of 139; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:153, and a VLCDR3 of the amino acid sequence of 153.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-22, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:139, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:153, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-23 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-23 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:140, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical amino acid sequence of the sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:154 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:140, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:154, and a VLCDR3 having an amino acid sequence of at least 90% identity thereto. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:140, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:154, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-23, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:140, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:154, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:140, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:154, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a sequence identical to SEQ ID NO: VHCDR1 comprising at least 90% identity to the amino acid sequence of l 30 comprising a sequence identical to SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:140, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:154, and a VLCDR3 of at least 90% identity to the amino acid sequence of seq id no.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:140, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:154, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-23, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:140, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:154, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-42 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-42 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:141 having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:155 has an amino acid sequence of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:141, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of 141; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:155 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:141, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:155, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-42, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:141, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:155, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:141, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity of VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:155, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:141, a VHCDR3 of at least 90% identity to the amino acid sequence of 141; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:155, and a VLCDR3 of at least 90% identity to the amino acid sequence of 155.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:141, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:155, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-42, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:141, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:155, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-3 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-3 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:142 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of the amino acid sequence; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:156 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:142, a VHCDR3 having an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:156 has an amino acid sequence having at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:142, a VHCDR3 of the amino acid sequence of seq id no; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:156, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-3, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:142, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:156, optionally comprising a VLCDR3 of 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:142, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:156, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VLCDR3.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:142, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:156, and a VLCDR3 of at least 90% identity to the amino acid sequence of 156.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:142, a VHCDR3 of the amino acid sequence of seq id no; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:156, and a VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-3, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:142, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:156, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

ADT1-7-61 and fragments and variants thereof

Certain embodiments relate to antibodies ADT1-7-61 and fragments and variants thereof.

For example, in some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:143 has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to the amino acid sequence of VHCDR3; and/or a light chain variable region comprising a sequence identical to SEQ ID NO:157 has an amino acid sequence of VLCDR3 of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a sequence identical to SEQ ID NO:143, a VHCDR3 having an amino acid sequence that is at least 90% identical; and/or a light chain variable region comprising a sequence identical to SEQ ID no:157 has an amino acid sequence of at least 90% identity. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:143, a VHCDR3 of the amino acid sequence; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:157, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-61, for example to provide an antibody or antigen binding fragment or variant thereof comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:143, optionally comprising a VHCDR3 of 1 or 2 amino acid substitutions; and/or a light chain variable region comprising a light chain sequence comprising SEQ ID NO:157, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

In some embodiments, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:131, and comprising a VHCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:143, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to VHCDR3; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to SEQ ID NO:145, and comprises a VLCDR2 comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to the amino acid sequence of SEQ ID NO:157, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical VLCDR3. In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a sequence identical to SEQ ID NO:130, comprising a VHCDR1 that is at least 90% identical to the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:143, a VHCDR3 of at least 90% identity to the amino acid sequence of seq id no; and/or

A light chain variable region comprising a sequence identical to SEQ ID NO:144, comprising a VLCDR1 having at least 90% identity to the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising at least 90% identity to the amino acid sequence of SEQ ID NO:157, and a VLCDR3 of at least 90% identity to the amino acid sequence of 157.

In one embodiment, an antibody or antigen-binding fragment or variant thereof is provided, comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:131 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:143, a VHCDR3 of the amino acid sequence; and

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:145 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:157, and VLCDR3 of the amino acid sequence of seq id no.

Amino acid substitutions may be made to provide variant antibodies derived from ADT1-7-61, for example to provide an antibody or antigen binding fragment or variant thereof comprising:

A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:130, a VHCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:131, a VHCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:143, a VHCDR3 optionally comprising 1 or 2 amino acid substitutions; and/or

A light chain variable region comprising a light chain sequence comprising SEQ ID NO:144, a VLCDR1 optionally comprising 1 or 2 amino acid substitutions, a polypeptide comprising the amino acid sequence of SEQ ID NO:145, a VLCDR2 optionally comprising 1 or 2 amino acid substitutions, and a polypeptide comprising the amino acid sequence of SEQ ID NO:157, optionally comprising 1 or 2 amino acid substitutions. Amino acid substitutions may be conservative amino acid substitutions.

Antibodies may alternatively consist of the indicated sequences (with or without amino acid substitutions).

Antibodies comprising heavy and/or light chain variable regions derived from ADT1-7

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 107 to 117 has or consists of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 119 to 129 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

Also provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 107 to 117, wherein the heavy chain variable region sequence is not SEQ ID NO: 106. And/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 119 to 129, wherein the light chain variable region sequence is not SEQ ID NO:118.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 112. 116 and 117 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 124. 128 and 129 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 107 to 117, or consists of a sequence having at least 90% identity to the sequence of the group consisting of seq id no; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 119 to 129, or a sequence having or consisting of at least 90% identity.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 112. 116 and 117, or a sequence having or consisting of at least 90% identity to the sequence of the group consisting of seq id no; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 124. 128 and 129 has or consists of a sequence having at least 90% identity.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

Provided herein is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 107 to 117 or a sequence consisting of said sequence; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 119 to 129 or a sequence consisting of said sequence.

Also provided is an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 112. 116 and 117 or a sequence consisting of said sequences; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID nos: 124. 128 and 129 or a sequence consisting of said sequences.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising a sequence identical to SEQ ID NO:107 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:119 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence;

b) Comprising a sequence identical to SEQ ID NO:108 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:120 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of 120;

c) Comprising a sequence identical to SEQ ID NO:109 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:121 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

d) Comprising a sequence identical to SEQ ID NO:110 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:122 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

e) Comprising a sequence identical to SEQ ID NO:111 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:123 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

f) Comprising a sequence identical to SEQ ID NO:112 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:124 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

g) Comprising a sequence identical to SEQ ID NO:113 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:125 or a VL consisting of an amino acid sequence at least 90% identical to the amino acid sequence of seq id no;

h) Comprising a sequence identical to SEQ ID NO:114 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:126 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence;

i) Comprising a sequence identical to SEQ ID NO:115 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:127 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no;

j) Comprising a sequence identical to SEQ ID NO:116 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:128 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence of seq id no; or (b)

K) Comprising a sequence identical to SEQ ID NO:117 or a VH consisting of or comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:129 or a VL consisting of an amino acid sequence that is at least 90% identical to the amino acid sequence,

Optionally, wherein the VH sequence is not SEQ ID NO:106 and/or VL sequences other than SEQ ID NO:118.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising SEQ ID NO:107 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:119 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

b) Comprising SEQ ID NO:108 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:120 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

c) Comprising SEQ ID NO:109 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:121 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

d) Comprising SEQ ID NO:110 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:122 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

e) Comprising SEQ ID NO:111 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:123 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

f) Comprising SEQ ID NO:112 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:124 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

g) Comprising SEQ ID NO:113 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:125 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

h) Comprising SEQ ID NO:114 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:126 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

i) Comprising SEQ ID NO:115 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:127 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions;

j) Comprising SEQ ID NO:116 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:128 or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions; or (b)

K) Comprising SEQ ID NO:117 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:129, or a VL consisting of said amino acid sequence, optionally comprising up to 5, up to 4, up to 3, up to 2 or 1 amino acid substitutions across two variable regions.

"Spans two variable regions" collectively refers to that an antibody may contain up to a specified number of substitutions when considering both heavy and light chain variable regions. Amino acid substitutions may be conservative amino acid substitutions. In some embodiments, substitution (if present) may occur anywhere in the variable region sequence. In preferred embodiments, substitution (if any) may be limited to the framework regions. Thus, in some embodiments, amino acid substitutions do not occur in the CDR sequences.

Optionally, the above antibodies retain the corresponding CDR sequences such that any variability in VH and VL sequences occurs in the framework regions.

In some embodiments, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) Comprising SEQ ID NO:107 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:119 or a VL consisting of said amino acid sequence;

b) Comprising SEQ ID NO:108 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:120 or a VL consisting of said amino acid sequence;

c) Comprising SEQ ID NO:109 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:121 or a VL consisting of said amino acid sequence;

d) Comprising SEQ ID NO:110 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:122 or a VL consisting of said amino acid sequence;

e) Comprising SEQ ID NO:111 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:123 or a VL consisting of said amino acid sequence;

f) Comprising SEQ ID NO:112 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:124 or a VL consisting of said amino acid sequence;

g) Comprising SEQ ID NO:113 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:125 or a VL consisting of said amino acid sequence;

h) Comprising SEQ ID NO:114 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:126 or a VL consisting of said amino acid sequence;

i) Comprising SEQ ID NO:115 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:127 or a VL consisting of said amino acid sequence;

j) Comprising SEQ ID NO:116 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:128 or a VL consisting of said amino acid sequence; or (b)

K) Comprising SEQ ID NO:117 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:129 or a VL consisting of said amino acid sequence.

Certain amino acid substitutions may be made to provide one or more variant antibodies as described herein.

In any embodiment involving a defined VH and/or VL sequence (e.g., defined as any VH and/or VL sequence having a certain percentage of identity and/or substitution), preferably the VH sequence is not SEQ ID NO:106 and the VL sequence is not SEQ ID NO:118.

Other antibodies derived from ADT1-7

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising SEQ ID NO:130 or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:131 or consists of said sequence;

c) An HCDR3 sequence comprising X ₁X₂YX₃X₄ AFDI (SEQ ID NO:183 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:132, a part of the material;

d) An LCDR1 sequence comprising SEQ ID NO:144 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:145 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₅X₆X₇X₈LX₉ T (SEQ ID NO:186 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:146;

wherein each of X ₁ to X ₉ is a naturally occurring amino acid.

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising SEQ ID NO:130 or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:13l or consists of said sequence;

c) An HCDR3 sequence comprising X ₁X₂YX₃X₄ AFDI (SEQ ID NO:184 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:132, a part of the material;

d) An LCDR1 sequence comprising SEQ ID NO:144 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:145 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₅X₆X₇X₈LX₉ T (SEQ ID NO:187 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:146;

Wherein X ₁ is selected from the group consisting of: D. i and V; x ₂ is selected from the group consisting of: d and S; x ₃ is selected from the group consisting of: n, E, D, Q and A; x ₄ is selected from the group consisting of: d and E; x ₅ is selected from the group consisting of: t and S; x ₆ is selected from the group consisting of: A. g, Y and S; x ₇ is selected from the group consisting of: s and D; x ₈ is selected from the group consisting of: t, E and G; and X ₉ is selected from the group consisting of: l and D. The antibody may further comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO: 189; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO: 190; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191, a sequence of 191; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192, a sequence of seq id no; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:193 and 194; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO: 195; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO: 196; and an LFR4 sequence, the LFR4 sequence comprising the sequence of SEQ ID NO: 197.

In one embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, is provided, the anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a) An HCDR1 sequence comprising SEQ ID NO:130 or consists of said sequence;

b) An HCDR2 sequence comprising SEQ ID NO:131 or consists of said sequence;

c) An HCDR3 sequence comprising X ₁X₂YX₃X₄ AFDI (SEQ ID NO:185 A) or consists of said sequence, wherein said HCDR3 sequence is not SEQ ID NO:132, a part of the material;

d) An LCDR1 sequence comprising SEQ ID NO:144 or consists of said sequence;

e) An LCDR2 sequence comprising SEQ ID NO:145 or consists of said sequence; and

F) An LCDR3 sequence comprising QQX ₅X₆X₇X₈LX₉ T (SEQ ID NO:188 A) or consists of said sequence, wherein said LCDR3 sequence is not SEQ ID NO:146;

Wherein X ₁ is selected from the group consisting of: d and V; x ₂ is selected from the group consisting of: d and S; x ₃ is selected from the group consisting of: D. q and A; x ₄ is selected from the group consisting of: d and E; x ₅ is S; x ₆ is selected from the group consisting of: a and Y; x ₇ is S; x ₈ is selected from the group consisting of: e and G; and X ₉ is selected from the group consisting of: l and D. The antibody may further comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO: 189; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO: 190; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191, a sequence of 191; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192, a sequence of seq id no; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO: 193; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO: 195; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO: 196; and an LFR4 sequence, the LFR4 sequence comprising the sequence of SEQ ID NO: 197.

Antibodies with sequence substitutions

The skilled artisan knows that the various amino acids have similar properties. One or more of such amino acids of a substance may typically be substituted with one or more other such amino acids without eliminating the desired activity of the substance.

Thus, the amino acids glycine, alanine, valine, leucine and isoleucine can generally be substituted for each other (amino acids having aliphatic side chains). Among these possible substitutions, it is preferable to use glycine and alanine for substitution with each other (because they have relatively short side chains), and valine, leucine and isoleucine for substitution with each other (because they have large aliphatic side chains, are hydrophobic). Other amino acids that may often be substituted for each other include: phenylalanine, tyrosine, and tryptophan (amino acids having aromatic side chains); lysine, arginine, and histidine (amino acids with basic side chains); aspartic acid and glutamic acid (amino acids with acidic side chains); asparagine and glutamine (amino acids with amide side chains); and cysteine and methionine (amino acids with sulfur-containing side chains).

A "conservative" amino acid substitution is one in which one amino acid residue is replaced by another amino acid residue of similar chemical structure, and is expected to have little effect on the function, activity, or other biological properties of the polypeptide. Such conservative substitutions are suitably substitutions in which one amino acid in the following group is substituted by another amino acid residue in the same group:

suitably, the hydrophobic amino acid residue is a non-polar amino acid. More suitably, the hydrophobic amino acid residue is selected from V, I, L, M, F, W or C. In some embodiments, the hydrophobic amino acid residue is selected from glycine, alanine, valine, methionine, leucine, isoleucine, phenylalanine, tyrosine, or tryptophan.

Thus, reference to a "conservative" amino acid substitution refers to an amino acid substitution in which one or more amino acids in the antibody sequence (e.g., in a CDR or in a VH or VL sequence) are replaced with another amino acid of the same class as shown above. Conservative amino acid substitutions may be preferred in the CDR regions to minimize adverse effects on antibody function. However, conservative amino acid substitutions may also occur in the framework regions. Thus, in some embodiments, any substitution in a CDR may be a conservative substitution, while a substitution in a framework region may be achieved by substituting a naturally occurring amino acid with another other naturally occurring amino acid.

Amino acid deletions or insertions may also be made relative to the amino acid sequences provided for the antibodies described herein. Thus, for example, amino acids which do not have a substantial effect on the activity of the polypeptide, or at least amino acids which do not eliminate such activity, may be deleted. Such deletions may be advantageous because the overall length and molecular weight of the polypeptide may be reduced while still retaining activity. This may reduce the amount of polypeptide required for a particular purpose-e.g., the dosage level may be reduced.

Amino acid changes relative to the sequences provided herein can be made using any suitable technique, for example, by using site-directed mutagenesis or solid state synthesis.

It is to be understood that amino acid substitutions or insertions within the scope of the invention may be made using naturally occurring or non-naturally occurring amino acids, although naturally occurring amino acids may be preferred. Whether natural or synthetic amino acids are used, it may be preferred that only L-amino acids be present.

Provided herein are various embodiments comprising optional amino acid substitutions of the provided sequences. In addition, in one embodiment of the invention, an antibody or antigen binding fragment thereof of the invention is provided comprising up to 10, suitably up to 5, or suitably up to 2 amino acid substitutions in the antibody binding domain or antigen binding domain. For example, in one embodiment of the invention, there is provided a vδ1 antibody or antigen binding fragment thereof, wherein said antibody or antigen binding fragment thereof comprises the 6 CDR regions ：ADT1-4-105、ADT1-4-107、ADT1-4-110、ADT1-4-112、ADT1-4-117、ADT1-4-19、ADT1-4-21、ADT1-4-31、ADT1-4-139、ADT1-4-4、ADT1-4-143、ADT1-4-53、ADT1-4-173、ADT1-4-2、ADT1-4-8、ADT1-4-82、ADT1-4-83、ADT1-4-3、ADT1-4-84、ADT1-4-86、ADT1-4-95、ADT1-4-1、ADT1-4-6、ADT1-4-138、ADT1-7-10、ADT1-7-15、ADT1-7-17、ADT1-7-18、ADT1-7-19、ADT1-7-20、ADT1-7-22、ADT1-7-23、ADT1-7-42、ADT1-7-3 and ADT1-7-61 of an antibody selected from the group consisting of an antibody or antigen binding fragment thereof having up to 10 amino acid substitutions, suitably up to 5 amino acid substitutions or up to 2 amino acid substitutions, over all of its CDR regions. In another embodiment of the invention, there is provided a vδ1 antibody or antigen binding fragment thereof, wherein said antibody or antigen binding fragment thereof comprises VH and VL sequences ：ADT1-4-105、ADT1-4-107、ADT1-4-110、ADT1-4-112、ADT1-4-117、ADT1-4-19、ADT1-4-21、AUT1-4-31、ADT1-4-139、ADT1-4-4、ADT1-4-143、ADT1-4-53、ADT1-4-173、ADT1-4-2、ADT1-4-8、ADT1-4-82、ADT1-4-83、ADT1-4-3、ADT1-4-84、ADT1-4-86、ADT1-4-95、ADT1-4-1、ADT1-4-6、ADT1-4-138、ADT1-7-10、ADT1-7-15、ADT1-7-17、ADT1-7-18、ADT1-7-19、ADT1-7-20、ADT1-7-22、ADT1-7-23、ADT1-7-42、ADT1-7-3 and ADT1-7-61 of an antibody selected from the group consisting of an antibody having up to 10 amino acid substitutions, suitably up to 5 amino acid substitutions or up to 2 amino acid substitutions, on its VH and VL sequences. In another embodiment of the invention, there is provided a vδ1 antibody or antigen binding fragment thereof, wherein said antibody or antigen binding fragment thereof is an antibody ：ADT1-4-105、ADT1-4-107、ADT1-4-110、ADT1-4-112、ADT1-4-117、ADT1-4-19、ADT1-4-21、ADT1-4-31、ADT1-4-139、ADT1-4-4、ADT1-4-143、ADT1-4-53、ADT1-4-173、ADT1-4-2、ADT1-4-8、ADT1-4-82、ADT1-4-83、ADT1-4-3、ADT1-4-84、ADT1-4-86、ADT1-4-95、ADT1-4-1、ADT1-4-6、ADT1-4-138、ADT1-7-10、ADT1-7-15、ADT1-7-17、ADT1-7-18、ADT1-7-19、ADT1-7-20、ADT1-7-22、ADT1-7-23、ADT1-7-42、ADT1-7-3 and ADT1-7-61 selected from the group consisting of said antibody or antigen binding fragment thereof having up to amino acid substitutions, suitably up to 5 amino acid substitutions or up to 2 amino acid substitutions. Substitutions are of course substitutions with reference to the original CDR or variable chain sequences of the starting antibody.

In some embodiments, one or more amino acid substitutions are located in one or more CDR regions. In other embodiments, one or more amino acid substitutions are located in the framework regions, i.e., in the variable heavy and light chains, but not in one or more CDR regions. In other embodiments, one or more amino acid substitutions may be located anywhere in the heavy chain variable region and/or the light chain variable region. In some embodiments, amino acid substitutions do not occur in the CDR sequences.

In some embodiments, amino acid substitutions do not adversely affect the binding specificity and/or affinity of the antibody. Thus, a variant antibody may have the same (or substantially the same) or superior functional characteristics as the antibody from which it was derived.

In some embodiments, amino acid substitutions may be made to increase the binding affinity of an antibody to a particular antigen. For example, in embodiments of the invention involving serine mutagenesis at light chain (variable region) position 74, substitutions may be made to increase the cross-reactivity of the antibody to cynomolgus homologs of the human antigen to which the antibody was made. Unlike the other substitutions described above, such substitutions may preferably be non-conservative. In some embodiments, the substitution can be a substitution of serine at position 74 with a nonpolar amino acid (e.g., a substitution with an amino acid selected from the group consisting of glycine, alanine, valine, methionine, leucine, and isoleucine). Or serine can be substituted with a non-human germline amino acid at position 74 (e.g., with an amino acid selected from the group consisting of arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine). In some embodiments, serine can be substituted with a non-germline and non-polar amino acid, i.e., an amino acid selected from the group consisting of glycine, valine, methionine, leucine, and isoleucine. In some embodiments, serine can be replaced with leucine.

Epitope(s)

Provided herein are antibodies (or antigen binding fragments thereof) that bind to an epitope of the vδ1 chain of γδ TCR. Such binding may optionally have an effect on γδ TCR activity, e.g. activation. The antibodies of the invention are specific for the vδ1 chain of γδ TCRs and do not bind to epitopes of other antigens, for example the vδ2 chain of γδ TCRs or the vδ3 chain of γδ TCRs. The antibodies of the invention may be considered agonistic antibodies, at least in terms of agonism to the vδ1 cells upon binding.

In one embodiment, the epitope may be an activating epitope of γδ T cells. An "activating" epitope may include, for example, stimulation of TCR function, such as cell degranulation, TCR down-regulation, cytotoxicity, proliferation, mobilization, increased survival or resistance to depletion, intracellular signaling, cytokine or growth factor secretion, a phenotypic change, or a change in gene expression. For example, binding of an activating epitope may stimulate expansion (i.e., proliferation) of a γδ T cell population, preferably a vδ1+ T cell population. Thus, these antibodies can be used to modulate γδ T cell activation, thereby modulating immune responses. Thus, in one embodiment, binding of the activating epitope down-regulates γδ TCR. In additional or alternative embodiments, binding of the activating epitope activates degranulation of γδ T cells. In another additional or alternative embodiment, binding of the activating epitope promotes γδ T cell mediated killing.

In some embodiments, the activating epitope of TRDV1 is an epitope that, upon binding by an antibody, results in down-regulation of the receptor and optionally activation of vδ1 cells. In some embodiments, the activating epitope is an epitope that upregulates expression of an activating marker, e.g., CD107a, CD25, CD69, and/or Ki67, on vδ1 cells. In some embodiments, the activating epitope is an epitope that upregulates expression of activating markers, such as CD107 and CD25, and optionally CD69 and/or Ki67 on vδ1 cells. In some embodiments, the upregulation of one or more activation markers (e.g., CD107 a) may be upregulation in the presence of cancer cells. In a preferred embodiment of the invention, the antibody binds to an activating epitope of TRDV a, in particular via the TRDV1 binding domain.

Since T cell receptors are often complexed with other proteins, down-regulation of T cell receptors via vδ1 antibody binding can cause down-regulation of other proteins associated with T cell receptors (i.e., binding of vδ1 antibody causes down-regulation of T cell receptor complex). For example, in some embodiments, the activating epitope of TRDV1 is an epitope that down-regulates the TCR/CD3 receptor complex upon binding. In this way, the antibodies of the invention may cause indirect downregulation of cell surface proteins that are not bound by the antibodies but are complexed with T cell receptors. Whereas the γδ1 chain-expressing T cells (i.e. vδ1 cells) represent only a few of the total T cell population, the antibodies of the present invention may be used to selectively (and indirectly) down-regulate proteins in the TCR complex, such as CD3, simply by down-regulating them in the vδ1 cells.

In some embodiments, the T cell receptor complex activating epitope is an epitope that down-regulates the T cell receptor complex after activation without down-regulating a CD3 molecule unassociated with the TRDV TCR complex.

The epitope preferably comprises at least one extracellular, soluble, hydrophilic or external portion of the vδ1 chain of γδ TCR.

In particular, the epitope does not comprise an epitope found in the hypervariable region of the vδ1 chain of γδ TCR (in particular CDR3 of the vδ1 chain). In a preferred embodiment, the epitope is within the non-variable region of the vδ1 chain of γδ TCR. It will be appreciated that such binding allows unique recognition of the vδ1 chain without limiting the highly variable TCR sequences (particularly CDR 3). In this way, various γδ TCR complexes recognizing antigens can be recognized solely by the presence of the vδ1 chain. Thus, it should be understood that any γδ TCR comprising a vδ1 chain may be recognized using an antibody or antigen binding fragment thereof as defined herein, regardless of the specificity of the γδ TCR. In one embodiment, the epitope comprises SEQ ID NO:272 and/or 35-90, such as a portion of the vδ1 chain that is not part of the CDR1 and/or CDR3 sequence. In one embodiment, the epitope does not comprise SEQ ID NO:272 amino acid residues within amino acid region 91-105 (CDR 3).

In some embodiments, the epitope comprises amino acids in the TRDV-1 CDR2 sequence.

In a similar manner to well-characterized αβt cells, γδ T cells utilize a diverse set of somatic rearrangement variable (V), diversity (D), linkage (J) and constant (C) genes, although γδ T cells contain fewer V, D and J segments than αβt cells. In one embodiment, the epitope bound by the antibody (or antigen binding fragment thereof) does not comprise an epitope found in the J region of the V.delta.1 chain (e.g., one of the four J regions encoded in the human delta.1 chain germline: SEQ ID NO:301 (J1 x 0) or 302 (J2 x 0) or 303 (J3 x 0) or 304 (J4 x 0)) or in the C region of the V.delta.1 chain (e.g., SEQ ID NO:305 (C1 x 0), which contains the C terminal proximal/transmembrane region). In one embodiment, the epitope bound by the antibody (or antigen binding fragment thereof) does not comprise an epitope found in the N-terminal leader sequence of the V.delta.1 chain (e.g., SEQ ID NO: 299). Thus, an antibody or fragment may only bind in the V region of the V.delta.1 chain (e.g., SEQ ID NO: 300). Thus, in one embodiment, the epitope consists of an epitope in the V region of the γδ TCR (e.g., amino acid residues 1-90 of SEQ ID NO: 272).

References to epitopes are made to vδ1 sequences derived from the sequences described in the following: luoma et al (2013) Immunity 39:1032-1042, and RCSB protein database entries: 3OMZ, shown as SEQ ID NO:272:

AQKVTQAQSSVSMPVRKAVTLNCLYETSWWSYYIFWYKQLPSKEMIFLIRQGSDEQNAKSGRYSVNFKKAAKSVALTISALQLEDSAKYFCALGESLTRADKLIFGKGTRVTVEPNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESS(SEQ ID NO：272).

SEQ ID NO:272 denotes a soluble TCR comprising V regions (also referred to as variable domains), D regions, J regions and TCR constant regions. The V region comprises amino acid residues 1-90, the D region comprises amino acid residues 91-104, the J region comprises amino acid residues 105-115, and the constant region (derived from T cell receptor alpha) comprises amino acid residues 116-209. Within the V region, CDR1 is defined as SEQ ID NO:272, amino acid residues 25-34, cd 2 is defined as SEQ ID NO:272, and CDR3 is defined as amino acid residues 50-54 of SEQ ID NO:272 (Xu et al, PNAS USA 108 (6): 2414-2419 (2011)).

Thus, according to one aspect of the present invention there is provided an isolated antibody or antigen binding fragment thereof which binds to an epitope of the variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) comprising one or more amino acid residues within the following amino acid region:

(i) SEQ ID NO:272 3-20; and/or

(Ii) SEQ ID NO:272 from 37 to 77.

In any aspect of the invention, there is provided an isolated antibody or antigen binding fragment thereof that binds to an epitope of the variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) comprising one or more amino acid residues within the following amino acid region:

(i) SEQ ID NO:272 3-20; and/or

(Ii) SEQ ID NO:272 from 37 to 77.

In another embodiment, the antibody or antigen binding fragment thereof additionally recognizes a polypeptide comprising SEQ ID NO:306 or a polymorphic V region consisting of said epitope. Thus, in defining the epitopes described herein, SEQ ID NO:272 and the polymorphic germline variation (amino acids 1-90 of SEQ ID NO: 306) may be considered interchangeable. The studies presented herein have demonstrated that the antibodies of the invention can recognize two variants of such germline sequences. For example, where it is indicated that an antibody or antigen binding fragment thereof as defined herein recognizes a polypeptide comprising SEQ ID NO:272, or an epitope consisting of or consisting of one or more amino acid residues within amino acid regions 1-24 and/or 35-90, which additionally recognizes the amino acid residues of SEQ ID NO:306 (i.e., at the same location).

In one embodiment, the antibody or antigen binding fragment thereof recognizes SEQ ID NO:272 from amino acid region 1 to 90. More specifically, in one embodiment, an antibody or antigen binding fragment thereof as defined herein recognizes a human germline epitope, wherein said germline encodes SEQ ID NO: alanine (a) or valine (V) at position 71 of 272.

In one embodiment, an epitope comprises or consists of one or more, e.g., two, three, four, five, six, seven, eight, nine, ten or more amino acid residues within the region.

In another embodiment, the epitope comprises SEQ ID NO:272, or consists of one or more (e.g., 5 or more, e.g., 10 or more) amino acid residues within amino acid region 3-20. In an alternative embodiment, the epitope comprises SEQ ID NO:272 (e.g., amino acid regions 50-54) within or consisting of one or more (e.g., 5 or more, e.g., 10 or more) amino acid residues. In another embodiment, the epitope comprises SEQ ID NO:272 (e.g., 5-20 or 3-17) and one or more (e.g., 5 or more, e.g., 10 or more) amino acid residues within amino acid region 37-77 (e.g., 62-77 or 62-69), e.g., 5 or more, e.g., 10 or more) amino acid residues.

It will be further understood that the antibody (or antigen binding fragment thereof) need not bind to all amino acids within a defined range. Such epitopes may be referred to as linear epitopes. For example, with a polypeptide comprising SEQ ID NO:272 amino acid residues within amino acid region 5-20 or an epitope consisting of said amino acid residues may bind only one or more amino acid residues within the range, e.g., amino acid residues at each end of the range (i.e., amino acids 5 and 20), optionally including amino acids within the range (i.e., amino acids 5, 9, 16 and 20).

In one embodiment, the epitope comprises SEQ ID NO:272, at least one of amino acid residues 3, 5, 9, 10, 12, 16, 17, 20, 37, 42, 50, 53, 59, 62, 64, 68, 69, 72 or 77, or consists thereof. In other embodiments, the epitope comprises a sequence selected from the group consisting of SEQ ID NOs: 272, one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve amino acids of or consisting of said amino acids.

In one embodiment, the epitope comprises SEQ ID NO:272 or consists of one or more amino acid residues within the following amino acid regions:

(i)3-17；

(ii)5-20；

(iii)37-53；

(iv)50-64；

(v)59-72；

(vi)59-77；

(vii) 62-69; and/or

(viii)62-77。

In other embodiments, the epitope comprises SEQ ID NO:272 or consists of one or more amino acid residues within the following amino acid regions: 5-20 and 62-77 (e.g., without limitation, embodiments involving antibodies derived from parental clone E07, e.g., affinity matured variants thereof); 50-64 (e.g., without limitation, embodiments involving antibodies derived from parental clone C08, e.g., affinity maturation variants thereof); 37-53 and 59-72 (e.g., without limitation, embodiments involving antibodies derived from parental clone G04, e.g., affinity matured variants thereof); 59-77 (e.g., without limitation, embodiments involving antibodies derived from parental clone C05, e.g., affinity maturation variants thereof); or 3-17 and 62-69 (e.g., without limitation, embodiments involving antibodies derived from parent clone E01, e.g., affinity matured variants thereof). In another embodiment, the epitope consists of SEQ ID NO:272, one or more amino acid residues within the following amino acid regions: 5-20 and 62-77;50-64;37-53 and 59-72;59-77; or 3-17 and 62-69.

In another embodiment, the epitope comprises SEQ ID NO:272 or consists of the following amino acid residues: 3. 5,9, 10, 12, 16, 17, 62, 64, 68 and 69, or, where appropriate, the amino acid sequence represented by SEQ ID NO:272, the following amino acid residue composition: 3. 5,9, 10, 12, 16, 17, 62, 64, 68 and 69 (e.g., without limitation, embodiments involving antibodies derived from parent clone E01, e.g., affinity matured variants thereof). In another embodiment, the epitope comprises SEQ ID NO:272 or consists of the following amino acid residues: 5. 9, 16, 20, 62, 64, 72 and 77, or, where appropriate, the amino acid sequence represented by SEQ ID NO:272, the following amino acid residue composition: 5. 9, 16, 20, 62, 64, 72 and 77 (e.g., without limitation, embodiments involving antibodies derived from parental clone E07, e.g., affinity matured variants thereof). In another embodiment, the epitope comprises SEQ ID NO:272 or consists of the following amino acid residues: 37. 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77, or, where appropriate, the amino acid sequence represented by SEQ ID NO:272, the following amino acid residue composition: 37. 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77 (e.g., without limitation, embodiments involving antibodies derived from parental clone G04, e.g., affinity matured variants thereof). In another embodiment, the epitope comprises SEQ ID NO:272 or consists of the following amino acid residues: 50. 53, 59, 62 and 64, or, where appropriate, the amino acid sequence represented by SEQ ID NO:272, the following amino acid residue composition: 50. 53, 59, 62 and 64 (e.g., without limitation, embodiments involving antibodies derived from parental clone C08, e.g., affinity matured variants thereof). In another embodiment, the epitope comprises SEQ ID NO:272 or consists of the following amino acid residues: 59. 60, 68 and 72, or, where appropriate, the sequences represented by SEQ ID NOs: 272, the following amino acid residue composition: 59. 60, 68 and 72 (e.g., without limitation, embodiments involving antibodies derived from parental clone C05, e.g., affinity matured variants thereof).

In one embodiment, the epitope comprises SEQ ID NO:272, or consist of one or more amino acid residues within amino acid regions 37-53 and/or 59-77. In another embodiment, the epitope consists of SEQ ID NO:272 from amino acid residues within amino acid regions 37-53 and 59-77. In another alternative embodiment, the epitope comprises SEQ ID NO:272 or one or more amino acid residues within or consisting of amino acid regions 37-53 or 59-77. Antibodies or antigen binding fragments thereof having such epitopes may have some or all of the sequence of G04, or such antibodies or antigen binding fragments thereof may be derived from G04. For example, an antibody or antigen binding fragment thereof having one or more CDR sequences of G04 or one or both of VH and VL sequences of G04 may bind such an epitope.

In one embodiment, the epitope comprises SEQ ID NO:272 from amino acid residues within or consisting of one or more amino acid residues within amino acid regions 5-20 and/or 62-77. In another embodiment, the epitope consists of SEQ ID NO:272 from amino acid regions 5-20 and 62-77. In another alternative embodiment, the epitope comprises SEQ ID NO:272 or 62-77, or consists of one or more amino acid residues within said amino acid region 5-20 or 62-77. Antibodies or antigen binding fragments thereof having such epitopes may have some or all of the sequence of E07, or such antibodies or antigen binding fragments thereof may be derived from E07. For example, an antibody or antigen-binding fragment thereof having one or more CDR sequences of E07 or one or both of the VH and VL sequences of E07 may bind such an epitope.

In one embodiment, the epitope comprises SEQ ID NO:272, or consists of one or more amino acid residues within amino acid regions 50-64. In another embodiment, the epitope consists of SEQ ID NO:272 of the amino acid sequence of amino acids 50 to 64. Antibodies or antigen binding fragments thereof having such epitopes may have some or all of the sequence of C08, or such antibodies or antigen binding fragments thereof may be derived from C08. For example, an antibody or antigen binding fragment thereof having one or more CDR sequences of C08 or one or both of VH and VL sequences of C08 may bind such an epitope.

In one embodiment, the epitope comprises SEQ ID NO:272 or one or more amino acid residues within amino acid regions 59-72. In another embodiment, the epitope consists of SEQ ID NO:272, and one or more amino acid residues within amino acid regions 59-72. Antibodies or antigen binding fragments thereof having such epitopes may have some or all of the sequence of C05, or such antibodies or antigen binding fragments thereof may be derived from C05. For example, an antibody or antigen-binding fragment thereof having one or more CDR sequences of C05 or one or both of VH and VL sequences of C05 may bind such an epitope.

In one embodiment, the epitope does not comprise SEQ ID NO:272 or consists of amino acid residues within amino acid regions 11-21. In one embodiment, the epitope does not comprise SEQ ID NO:272 or consists of amino acid residues within amino acid regions 21-28. In one embodiment, the epitope does not comprise SEQ ID NO:272, or consists of amino acid residues within amino acid regions 59 and 60. In one embodiment, the epitope does not comprise SEQ ID NO:272 or consists of amino acid residues within amino acid regions 67-82.

The epitope of the affinity matured antibody is typically the same as the epitope identified herein for the parental clone. For those epitopes on cynomolgus TRDV1, the epitope positions of the affinity matured antibodies are typically the same positions as the epitopes identified for the corresponding parental clones. Reference to "position" is necessary because the skilled artisan will appreciate that the identity of some amino acids in an epitope differs from human TRDV1. Despite these differences between human and cynomolgus TRDV1, such antibodies are still capable of specifically binding to both antigens.

In one embodiment, the epitope is different from the epitope to which a commercially available anti-V.delta.1 antibody, such as TS-1 or TS8.2, binds. As described in WO2017197347, when the delta 1 chain included the V delta 1 J1 and V delta 1 J2 sequences but did not include the V delta 1 J3 chain, binding of TS-1 and TS8.2 to the soluble TCR was detected, indicating that binding of TS-1 and TS8.2 involves critical residues in the delta J1 and delta J2 regions.

References herein to "within … …" include the endpoints of the defined range. For example, "within amino acid region 5-20" refers to all amino acid residues from residue 5 (and including residue 5) up to residue 20 (and including residue 20).

Various techniques for establishing the epitope to which an antibody binds are known in the art. Exemplary techniques include, for example, conventional cross-blocking assays, alanine scanning mutation analysis, peptide blot analysis, peptide cleavage analysis crystallography and NMR analysis. In addition, methods such as epitope excision, epitope extraction and chemical modification of antigens may also be employed. Another method that may be used to identify amino acids within a polypeptide that interact with an antibody is hydrogen/deuterium exchange detected by mass spectrometry (as described in example 9). In general, the hydrogen/deuterium exchange method involves deuterium labeling a protein of interest, and then binding the antibody to the deuterium labeled protein. Next, the protein/antibody complex is transferred into water and exchangeable protons within the amino acids protected by the antibody complex undergo deuterium-to-hydrogen reverse exchange at a slower rate than exchangeable protons within amino acids not part of the interface. Thus, amino acids forming part of the protein/antibody interface may retain deuterium and thus exhibit relatively higher quality than amino acids not included in the interface. After dissociation of the antibody, the target protein is subjected to protease cleavage and mass spectrometry analysis, revealing deuterium labeled residues corresponding to the particular amino acids with which the antibody interacts.

The antibody and antigen binding fragments thereof suitably bind specifically to human TRDV1 (polymorphic variants of SEQ ID NO:272 and SEQ ID NO: 306) and to orthologs in cynomolgus monkey, cynomolgus monkey TRDV1 (SEQ ID NO:308, see also UniProtKB-G7P9S6 (G7P9S6_ MACFA)). In some embodiments, the antibody or antigen binding fragment thereof is a G04-derived antibody that specifically binds to both human and cynomolgus TRDV 1. It is assumed that the epitope to which the antibodies of the invention bind (e.g. but not limited to those epitopes that bind in the region within amino acid residues 37 to 77 of any of SEQ ID NOs 272, 306 or 308) may be particularly advantageous as it allows for the provision of anti-vδ1 antibodies specific for vδ1 (i.e. do not bind to similar antigens, such as vδ2 or vδ3) but also provides for cross-reactivity with polymorphic variants of vδ1 (i.e. TRDV SEQ ID NOs 272 and TRDV SEQ ID NOs 306, despite the presence of a polymorphism at residue position 20 and identified as possible contact residues or as proximity to the identified contact residues of certain antibodies) and provides cross-reactivity between human and cynomolgus monkey vδ1 (although residues 42, 50, 54, 59, 60, 68, 73, 75 and 76 occurring within the region of amino acid residues 37 to 77 of SEQ ID NOs 272 and 308 are different between human and TRDV1 sequences).

Epitope of ADT1-4 derived antibodies

The ADT1-4 derived antibody binds to the same or substantially the same epitope as the ADT1-4 parent antibody. Thus, in some embodiments, particularly those involving antibodies derived from or related to the ADT1-4 parent antibody, the anti-vδ1 antibody or antigen binding fragment binds a polypeptide comprising SEQ ID NO:272 (or SEQ ID NO: 306) within amino acid regions 37 to 77, for example one or more amino acid residues within amino acid regions 37 to 53 and/or 59 to 77 or an epitope consisting of said amino acid residues. In some embodiments, the epitope comprises SEQ ID NO:272 (or SEQ ID NO: 306) amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77 or consists of said amino acid residues. In some embodiments, the epitope consists of SEQ ID NO:272, amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77 (or SEQ ID NO: 306).

ADT1-4 derived antibodies also bind to an epitope of the cynomolgus monkey variable delta 1 (V delta 1) chain, also known as cynomolgus monkey TRDV1 (SEQ ID NO: 308), of the gamma delta T Cell Receptor (TCR). Thus, in some embodiments, particularly those involving antibodies derived from or related to the G04 parent antibody, the anti-vδ1 antibody or antigen binding fragment binds a polypeptide comprising SEQ ID NO:308, e.g., one or more amino acid residues within amino acid regions 37-53 and/or 59-77 or an epitope consisting of said amino acid residues. In some embodiments, the epitope comprises SEQ ID NO:308 or consists of amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77. In some embodiments, the epitope consists of SEQ ID NO:308, amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

In view of the cross-reactivity of antibodies derived from ADT1-4 in particular, and the ability of the antibodies disclosed herein to bind polymorphic variants of TRDV1, in some embodiments, in particular those embodiments involving antibodies derived from or related to the G04 parent antibody, the anti-vδ1 antibody or antigen binding fragment binds a polypeptide comprising SEQ ID NO: 272. 306 and 308, e.g., one or more amino acid residues within amino acid regions 37-77, e.g., amino acid regions 37-53 and/or 59-77, or an epitope consisting of said amino acid residues. In some embodiments, the epitope comprises SEQ ID NO: 272. 306 and 308, or consists of amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77. In some embodiments, the epitope consists of SEQ ID NO: 272. 306 and 308, and amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

Given the location of amino acid variants between these sequences from both human and cynomolgus species, it is surprising to provide antibodies comprising or consisting of epitopes as described above but providing cross-reactivity between human and cynomolgus TRDV1 sequences.

Epitope of ADT1-7 derived antibodies

Antibodies derived from ADT1-7 bind the same or substantially the same epitope as the ADT1-7 parent antibody. Thus, in some embodiments, particularly those involving antibodies derived from or related to the ADT1-7 parent antibody, the anti-vδ1 antibody or antigen binding fragment binds a polypeptide comprising SEQ ID NO:272 (or SEQ ID NO: 306) one or more amino acid residues within amino acid regions 5-20 and/or 62-77 or an epitope consisting of said amino acid residues. In some embodiments, the epitope comprises SEQ ID NO:272 (or SEQ ID NO: 306) amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77 or consists of said amino acid residues. In some embodiments, the epitope consists of SEQ ID NO:272 (or SEQ ID NO: 306) amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77.

Frames and other sequences

Suitably, the VH and VL regions of an antibody or antigen-binding fragment of the invention each comprise four framework regions (FR 1-FR 4). In one embodiment, the antibody or antigen binding fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and/or FR 4) comprising a sequence having at least 80% sequence identity to a framework region in any one of: SEQ ID NO:2 to 25 (e.g. in the case of a light chain variable sequence derived from G04), SEQ ID NO:27 to 50 (e.g. in the case of heavy chain variable sequences derived from G04), SEQ ID NO:107 to 117 (for example in the case of a light chain variable sequence derived from E07) or SEQ ID NO:119 to 129 (e.g. in the case of a light chain variable sequence derived from E07). In one embodiment, the antibody or antigen binding fragment thereof comprises a framework region (e.g., FRI, FR2, FR3, and/or FR 4) comprising a sequence having at least 90%, e.g., at least 95%, 97%, or 99% sequence identity to a framework region in any one of: SEQ ID NO:2 to 25 (e.g. in the case of a light chain variable sequence derived from G04), SEQ ID NO:27 to 50 (e.g. in the case of heavy chain variable sequences derived from G04), SEQ ID NO:107 to 117 (for example in the case of a light chain variable sequence derived from E07) or SEQ ID NO:119 to 129 (e.g. in the case of a light chain variable sequence derived from E07). In one embodiment, the antibody or antigen binding fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and/or FR 4) comprising a sequence in any one of: SEQ ID NO:2 to 25 (e.g. in the case of a light chain variable sequence derived from G04), SEQ ID NO:27 to 50 (e.g. in the case of heavy chain variable sequences derived from G04), SEQ ID NO:107 to 117 (for example in the case of a light chain variable sequence derived from E07) or SEQ ID NO:119 to 129 (e.g. in the case of a light chain variable sequence derived from E07). In one embodiment, the antibody or antigen binding fragment thereof comprises a framework region (e.g., FR1, FR2, FR3, and/or FR 4) consisting of a sequence in any one of: SEQ ID NO:2 to 25 (e.g. in the case of a light chain variable sequence derived from G04), SEQ ID NO:27 to 50 (e.g. in the case of heavy chain variable sequences derived from G04), SEQ ID NO:107 to 117 (for example in the case of a light chain variable sequence derived from E07) or SEQ ID NO:119 to 129 (e.g. in the case of a light chain variable sequence derived from E07).

In some embodiments, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: HFR1 (i.e., heavy chain framework 1 region) sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or 178 or consists of said sequence; and/or LFR4 sequences, said LFR4 sequences comprising SEQ ID NO: 179. 180, 181 or 182 or a sequence consisting of said sequences.

In some embodiments, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: HFR1 (i.e., heavy chain framework 1 region) sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or consists of said sequence; and/or LFR4 sequences, said LFR4 sequences comprising SEQ ID NO: 179. 180, 181 or 182 or a sequence consisting of said sequences.

In some embodiments, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: HFR1 (i.e., heavy chain framework 1 region) sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or consists of said sequence; and/or LFR4 sequences, said LFR4 sequences comprising SEQ ID NO:179 or 181 or consists of said sequence.

In some embodiments, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:189 or consists of said sequence; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:190 or consists of said sequence; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191 or consists of said sequence; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192 or consists of said sequence; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:193 and 194 or consists of said sequences; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:195 or consists of said sequence; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:196 or consists of said sequence; and an LFR4 sequence, the LFR4 sequence comprising the sequence of SEQ ID NO:197 or consists of said sequence.

In some embodiments, an anti-vδ1 antibody or antigen binding fragment thereof may comprise: an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:189 or consists of said sequence; an HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:190 or consists of said sequence; an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191 or consists of said sequence; an HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192 or consists of said sequence; an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:193 or consists of said sequence; an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:195 or consists of said sequence; an LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:196 or consists of said sequence; and an LFR4 sequence, the LFR4 sequence comprising the sequence of SEQ ID NO:197 or consists of said sequence.

For fragments comprising both VH and VL regions, these regions may be associated covalently (e.g., via disulfide bonds or linkers) or non-covalently. The antibody fragments described herein may comprise scFv, i.e. fragments comprising a VH region and a VL region joined by a linker. In one embodiment, the VH region and VL region are joined by a (e.g., synthetic) polypeptide linker. The polypeptide linker may comprise (Gly ₄Ser)_n linker, where n = 1 to 8, e.g. 2, 3, 4, 5 or 7. The polypeptide linker may comprise [ (Gly ₄Ser)_n(Gly₃AlaSer)_m]_p linker, where n = 1 to 8, e.g. 2, 3, 4, 5 or 7, m = 1 to 8, e.g. 0, 1, 2 or 3, and p = 1 to 8, e.g. 1, 2 or 3. In another embodiment, the linker comprises SEQ ID NO:291. In another embodiment, the linker consists of SEQ ID NO: 291).

Those skilled in the art will appreciate that scFv constructs can be designed and made to include N-terminal and C-terminal modifications to aid translation, purification, and detection. For example, at the N-terminus of the scFv sequence, additional methionine and/or alanine amino acid residues may be included prior to normalizing the VH sequence (e.g., start QVQ or EVQ). At the C-terminus (i.e., the C-terminus of the canonical mature VL domain sequence ending according to the IMGT definition), additional sequences may be included, such as (i) partial sequences of the constant domain and/or (ii) additional synthetic sequences, including tags, such as His tags and Flag tags, to aid in purification and detection (e.g., tags of any one of SEQ ID NOS: 292-295).

The antibodies may be in any format, as described herein. In a preferred embodiment, the antibody is in the IgG1 (e.g., human IgG 1) format (i.e., the antibody is a human IgG1 antibody).

In some embodiments, the antibody or antigen binding fragment thereof comprises: a light chain constant region comprising the amino acid sequence of SEQ ID NO:296 or SEQ ID NO:307 (or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID No. 296 or SEQ ID No. 307); and/or a heavy chain constant region comprising the amino acid sequence of SEQ ID NO:297 or SEQ ID NO:298 (or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID No. 297 or SEQ ID No. 298). In some embodiments, effector function of the heavy chain constant region may be reduced or lost (effector function disabling mutation). Suitable mutations which attenuate effector functions are known to the skilled worker. For example, the L235A and/or G237A mutation ("LAGA") or the L234A and/or L235A mutation ("LALA") according to EU numbering. For example, in one embodiment, the antibody or antigen binding fragment thereof comprises a polypeptide comprising SEQ ID NO:296 and/or a light chain constant region comprising the sequence of SEQ ID NO:297 or SEQ ID NO: 298.

Competing antibodies

In one embodiment, the antibody binds to the same or substantially the same epitope as the antibody or antigen binding fragment thereof as defined herein, or competes with the antibody or antigen binding fragment thereof as defined herein. Whether an antibody binds to the same epitope as a reference anti-vδ1 antibody or competes for binding with a reference anti-vδ1 antibody can be readily determined by using conventional methods known in the art. For example, to determine whether a test antibody binds to the same epitope as a reference anti-vδ1 antibody of the present invention, the reference antibody is allowed to bind to vδ1 protein or peptide under saturation conditions. Next, the ability of the test antibodies to bind to the V.delta.1 chain was assessed. If the test antibody is capable of binding to vδ1 after saturation binding to the reference anti-vδ1 antibody, it may be concluded that the test antibody binds a different epitope than the reference anti-vδ1 antibody. On the other hand, if the test antibody cannot bind to the vδ1 chain after saturation binding with the reference anti-vδ1 antibody, the epitope to which the test antibody binds may be the same as the epitope to which the reference anti-vδ1 antibody of the present invention binds.

The invention also includes anti-vδ1 antibodies that compete for binding to vδ1 with antibodies as defined herein or antigen binding fragments thereof or antibodies having CDR sequences of any exemplary antibodies described herein. For example, competition assays can be performed with the antibodies of the invention to determine which proteins, antibodies, and other antagonists compete with the antibodies of the invention for vδ1 chain binding and/or share epitopes. These assays are readily known to those skilled in the art; they evaluate competition between antagonists or ligands for a limited number of binding sites (e.g., vδ1) on proteins. The antibody (or antigen binding fragment thereof) is immobilized or insoluble either before or after competition and the sample bound to the vδ1 chain is separated from the unbound sample, for example by decantation (where the antibody is pre-insoluble) or by centrifugation (where the antibody precipitates after competition reaction). In addition, competitive binding can be determined by whether the function is altered by binding or not binding of the antibody to the protein, e.g., whether the antibody molecule inhibits or enhances, e.g., the enzymatic activity of the label. ELISA and other functional assays can be used as known in the art and described herein.

Two antibodies bind to the same or overlapping epitope if each of the two antibodies competitively inhibits (blocks) the binding of the other antibody to the target antigen. That is, a 1, 5, 10, 20, or 100-fold excess of one antibody inhibits the binding of the other by at least 50%, but preferably 75%, 90%, or even 99%, as measured in a competitive binding assay. Alternatively, two antibodies have the same epitope if substantially all amino acid mutations in the target antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody.

Additional routine experimentation (e.g., peptide mutation and binding analysis) can then be performed to confirm that no test antibody binding is observed whether it is actually due to the same epitope as the reference antibody binding, or whether steric blocking (or another phenomenon) is responsible for the non-observed binding. Such experiments can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art.

Antibody binding and pharmacological Properties of antibodies

Antibodies of the invention may have advantageous binding and/or pharmacological properties, for example as described below.

Binding affinity (K _D)

In general, affinity matured clones have a higher affinity for their antigen than the parent clone. For example, the affinity for human TRDV1 (SEQ ID NO 272 or 306) will be at least 20%, at least 30%, at least 40%, at least 50%, at least 100% or at least 500% higher than the parent antibody.

The antibodies of the invention, or antigen-binding fragments thereof, may bind to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50nM.

An antibody or antigen-binding fragment thereof may be further defined as binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 10nM, preferably less than about 5 nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-4 lineage or derived from the ADT1-4 lineage, an antibody or antigen-binding fragment thereof of the invention can bind to human TRDV1 at a K _D of less than about 100nM, preferably less than about 50 nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-4 lineage (except for ADT 1-4-138) or derived from the ADT1-4 lineage (except for ADT 1-4-138), an antibody or antigen-binding fragment thereof of the invention may bind to human TRDV1 at a K _D of less than about 10nM, preferably less than about 5 nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-4 lineage members ADT1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112, ADT1-4-143, and/or ADT1-4-1, an antibody or antigen binding fragment thereof of the invention may bind to human TRDV at a K _D of less than about 10nM, preferably less than about 1 nM.

The antibodies of the invention, or antigen binding fragments thereof, may bind to cynomolgus monkey TRDV1 (SEQ ID NO 308) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-4 lineage or derived from the ADT1-4 lineage, an antibody or antigen-binding fragment thereof of the invention can bind to cynomolgus monkey TRDV1 at K _D of less than about 100nM, preferably less than about 50 nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-4 lineage members ADT1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112, ADT1-4-143, and/or ADT1-4-1, an antibody or antigen binding fragment thereof of the invention can bind to cynomolgus monkey TRDV1 at a K _D of less than about 50 nM.

In some embodiments, such as those antibodies associated with the ADT1-4 lineage or derived from the ADT1-4 lineage, the antibodies of the invention, or antigen binding fragments thereof, may bind to human TRDV1 with a K _D of less than about 100nM, preferably less than about 50nM, and to cynomolgus monkey TRDV1 with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-4 lineage (except for ADT 1-4-138) or derived from the ADT1-4 lineage (except for ADT 1-4-138), an antibody or antigen-binding fragment thereof of the invention can bind to human TRDV with a K _D of less than about 10nM and to cynomolgus monkey TRDV1 with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100 nM. Preferably in this embodiment, an antibody or antigen binding fragment thereof of the invention may bind to human TRDV1 with a K _D of less than about 5nM and to cynomolgus monkey TRDV1 with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 50 nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-4 lineage members ADT1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112, ADT1-4-143, and/or ADT1-4-1, an antibody or antigen binding fragment thereof of the invention can bind to human TRDV at a K _D of less than about 10nM and to human TRDV1 at a K _D of less than 50 nM. Preferably in this embodiment, an antibody or antigen binding fragment thereof of the invention may bind to human TRDV with a K _D of less than about 1nM and to cynomolgus monkey TRDV1 with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 50 nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-7 lineage or derived from the ADT1-7 lineage, an antibody or antigen-binding fragment thereof of the invention can bind to human TRDV1 at a K _D of less than about 10nM, preferably less than about 5 nM.

In some embodiments, the binding affinity of an antibody or antigen binding fragment thereof is determined by coating the antibody or antigen binding fragment thereof directly or indirectly (e.g., by capture with anti-human IgG Fc) onto the surface of a sensor (e.g., an amine high capacity chip or equivalent), wherein the target bound by the antibody or antigen binding fragment thereof (i.e., the vδ1 chain of γδ TCR) flows through the chip to detect binding. Suitably, MASS-2 instrumentation (also referred to as SIERRA SPR-32) is used in 30. Mu.l/min PBS+0.02% Tween20 running buffer at 25 ℃.

Other assays useful for defining antibody function are described herein. For example, an antibody or antigen binding fragment thereof described herein can be assessed by γδ TCR engagement, e.g., measuring down-regulation of γδ TCR after antibody binding. Surface expression of γδ TCRs after application of antibodies or antigen binding fragments thereof (optionally present on the cell surface) can be measured, for example, by flow cytometry. Antibodies or antigen binding fragments thereof described herein can also be assessed by measuring γδ T cell degranulation. For example, expression of CD107a (a marker of cell degranulation) can be measured, e.g., by flow cytometry, after application of the antibody or antigen binding fragment thereof (optionally presented on the cell surface) to γδ T cells. The antibodies or antigen binding fragments thereof described herein can also be evaluated by measuring γδ T cell mediated killing activity (to test whether the antibodies have an effect on γδ T cell killing activity). For example, the target cells may be incubated with γδ T cells in the presence of the antibody or antigen binding fragment thereof (optionally present on the cell surface). After incubation, the cultures may be stained with a cell viability dye to distinguish live and dead target cells. The proportion of dead cells can then be measured, for example, by flow cytometry.

Inhibitory concentrations (IC 50 and IC 90)

TCR Down-Regulation

TCR downregulation can be measured according to the assays described herein. For example, antibodies to be tested may be incubated with γδ T cell cultures at different concentrations and measured for down-regulation. If cell killing (e.g., THP-1 cell killing) is measured, γδ T cells are co-cultured with a suitable cell line (e.g., THP-1 cells). TCR downregulation can be measured by flow cytometry. Cell killing may be achieved by any suitable means, for example by flow cytometry. For initial TCR downregulation studies using parent antibodies (e.g., ADT1-7 and ADT 1-4), the TCR downregulation assay involved 'loading' the antibodies onto fcγ receptor+thp-1 cells (see example 1, example 6, and table 5). Thus, in this case, the antibodies are presented on the cell surface prior to co-incubation with γδ T cells. Thus, this loading provides the greatest opportunity to exploit the impact of cross-linking on TCR engagement. In addition to loading the antibody onto Fc receptor + cells, alternative similar methods of presenting the antibody on a solid surface may include pre-incubating the antibody on a plate (so-called plate binding), or using carrier beads to present the antibody. In all such assays, once presented on a solid surface, the technical impact that antibodies impart upon binding to a receptor of interest (γδ T cell receptor in this example) is typically studied and measured, and it is common to present antibodies in this manner, particularly when exploring the impact of antibodies binding to immune cell receptor targets and complexes, such as antibodies binding to targets like CD3, CD28, etc.

In contrast, for affinity matured antibodies of the invention as described herein, the inventors want to most fully profile and compare the effects of affinity maturation, and therefore the inventors explored the ability of these antibodies in a more 'soluble' TCR down-regulation assay format. Assessing the effect in solution may also be more physiologically relevant. For these reasons, and unless otherwise indicated, in all cell-based TCR downregulation experiments, a soluble assay format was explored in which the effect of affinity matured antibodies, multispecific antibodies, or fragments thereof was measured or characterized and compared to the parent antibody (see, e.g., fig. 35). The differences in IC50 using this more stringent solubility assay are summarized below, wherein TCR downregulation of EC50 values for the exemplary parent molecule (ADT 1-4) is summarized by two methods.

TCR down-regulation: 'present' contrast 'soluble' detection format:

ADT1-4 IC50 (loaded on THP-1 cells): 0.01-0.05 ug/ml=0.065 nM to 0.325nM

(See example 1, example 6 and Table 5)

Comparison

ADT1-4 ic50 (solubility assay, added to solution) =38.28 nM

(See FIG. 35F)

The antibodies or antigen binding fragments thereof of the invention may have an IC50 for TCR down-regulation of less than about 50nM. Preferably, the IC50 is less than about 10nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-4 lineage, an antibody or antigen binding fragment thereof of the invention can have an IC50 for TCR downregulation of less than about 50nM and/or an IC90 for TCR downregulation of less than about 100nM. Preferably, the IC50 is less than about 10nM and the IC90 is less than about 50nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with the ADT1-4 lineage (except for ADT 1-4-138) or derived from the ADT1-4 lineage (except for ADT 1-4-138), an antibody or antigen binding fragment thereof of the invention can have an IC50 for TCR down-regulation of less than about 1nM and/or an IC90 for TCR down-regulation of less than about 10nM. Preferably, the IC50 is less than about 0.5nM and the IC90 is less than about 5nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-4 lineage members ADT1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112, ADT1-4-143, and/or ADT1-4-1, an antibody or antigen binding fragment thereof of the invention can have an IC50 for TCR down-regulation of less than about 1nM and/or an IC90 for TCR down-regulation of less than about 10nM. Preferably, the IC50 is less than about 0.5nM and the IC90 is less than about 5nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-7 lineage or derived from the ADT1-7 lineage, an antibody or antigen-binding fragment thereof of the invention allows for TCR down-regulation of an IC50 of less than about 50nM. Preferably, the IC50 is less than about 10nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-7 lineage members ADT1-7-20 or ADT1-7-3, an antibody or antigen binding fragment thereof of the invention can have an IC50 for TCR downregulation of less than about 5nM and/or an IC90 for TCR downregulation of less than about 10nM. Preferably, the IC50 is less than about 10nM and the IC90 is less than about 5nM.

THP-1 cell killing

Cell killing may be measured according to the assays described herein. For example, the antibody to be tested may be incubated with co-cultures of γδ T cells and tumor cells (e.g., THP-1 cells) at varying concentrations. Cell killing may be measured by any suitable means, for example by flow cytometry.

The antibodies or antigen binding fragments thereof of the invention may have an IC50 of less than about 10nM for killing THP-1 cells and/or an IC90 of less than about 50nM for killing THP-1 cells. Preferably, the IC50 is less than about 5nM and the IC90 is less than about 50nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-4 lineage or derived from the ADT1-4 lineage, an antibody or antigen-binding fragment thereof of the invention can have an IC50 of less than about 10nM to kill THP-1 cells and/or an IC90 of less than about 50nM to kill THP-1 cells. Preferably, the IC50 is less than about 5nM and the IC90 is less than about 50nM.

In some embodiments, for example, an antibody or antigen-binding fragment thereof associated with the ADT1-7 lineage or derived from the ADT1-7 lineage, an antibody or antigen-binding fragment thereof of the invention can have an IC50 of less than about 5nM to kill THP-1 cells and/or an IC90 of less than about 50nM to kill THP-1 cells. Preferably, the IC50 is less than about 1nM and the IC90 is less than about 50nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with or derived from ADT1-7 lineage members ADT1-7-20 or ADT1-7-3, an antibody or antigen binding fragment thereof of the invention can have an IC50 of less than about 5nM for killing THP-1 cells and/or an IC90 of less than about 10nM for killing THP-1 cells. Preferably, the IC50 is less than about 1nM and the IC90 is less than about 5nM.

Of course, the advantageous pharmacological properties of the antibodies may be combined such that the antibodies exhibit advantageous K _D and advantageous IC50 and/or IC90 values for various test properties.

For example, in some embodiments, an antibody or antigen binding fragment thereof of the invention may:

Binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

Optionally binding to cynomolgus monkey TRDV1 (SEQ ID NO 308) with a binding affinity (K _D, e.g. measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

IC50 for down-regulating TCR of less than about 50nM (preferably less than about 10 nM);

IC50 for killing THP-1 cells of less than about 10nM (preferably less than about 5 nM) and

IC90 to kill THP-1 cells is less than about 50nM.

In some embodiments, particularly those antibodies, or antigen binding fragments thereof, that relate to ADT1-4 lineages, may:

bind to human TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

Binding to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

IC50 for TCR downregulation of less than about 50nM (preferably less than about 10 nM) and IC90 for TCR downregulation of less than about 100nM (less than about 50 nM);

IC50 for killing THP-1 cells of less than about 10nM (preferably less than about 5 nM)

IC90 to kill THP-1 cells is less than about 50nM.

In some embodiments, for example, antibodies or antigen-binding fragments thereof associated with the ADT1-4 lineage (except for ADT 1-4-138) or derived from the ADT1-4 lineage (except for ADT 1-4-138), an antibody or antigen-binding fragment of the invention can:

Bind to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

Binding to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and IC90 for TCR downregulation of less than about 10nM (less than about 5 nM);

IC50 for killing THP-1 cells of less than about 10nM (preferably less than about 5 nM)

IC90 to kill THP-1 cells is less than about 50nM.

In some embodiments, for example, an antibody or antigen binding fragment thereof associated with ADT1-4 lineage members ADT1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112, ADT1-4-143, and/or ADT1-4-1 or derived from said ADT1-4 lineage members, an antibody or antigen binding fragment thereof of the invention can:

Bind to human TRDV1 with a K _D of less than about 10nM (preferably less than about 1 nM);

binding to cynomolgus monkey TRDV1 with a K _D of less than about 50 nM;

IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and IC90 for TCR downregulation of less than about 10nM (less than about 5 nM);

IC50 for killing THP-1 cells of less than about 10nM (preferably less than about 5 nM)

IC90 to kill THP-1 cells is less than about 50nM.

In some embodiments, particularly those antibodies, or antigen binding fragments thereof, that relate to ADT1-7 lineages, may:

Bind to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

IC50 for down-regulating TCR of less than about 50nM (preferably less than about 10 nM);

IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM)

IC90 to kill THP-1 cells is less than about 50nM.

In some embodiments, particularly those antibodies, antibodies or antigen binding fragments thereof, that relate to ADT1-7 lineages ADT1-7-20 or ADT1-7-3, may:

Bind to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

IC50 for down-regulating TCR of less than about 5nM (preferably less than about 1 nM);

IC90 to down-regulate TCR of less than about 10nM (preferably less than about 5 nM);

IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM)

IC90 to kill THP-1 cells is less than about 10nM (preferably less than about 5 nM).

Pharmacological Properties of ADT1-4 lineage antibodies

The antibody to ADT1-4-105 or the antibody derived from ADT1-4-105 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-107 or an antibody derived from ADT1-4-107 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-110 or the antibody derived from ADT1-4-110 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

Antibodies to ADT1-4-112 or antibodies derived from ADT1-4-112 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50nM. Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

The antibody to ADT1-4-117 or the antibody derived from ADT1-4-117 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-19 or the antibody derived from ADT1-4-19 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV of less than about 1nM (preferably less than about 0.5 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

The antibody to ADT1-4-21 or the antibody derived from ADT1-4-21 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

The antibody to ADT1-4-31 or the antibody derived from ADT1-4-31 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM for killing THP-1 cells.

The antibody to ADT1-4-139 or an antibody derived from ADT1-4-139 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

Antibodies to ADT1-4-4 or antibodies derived from ADT1-4-4 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

Antibodies to ADT1-4-143 or antibodies derived from ADT1-4-143 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage identity) may have K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM) and/or K _D to cynomolgus monkey TRDV1 of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM for killing THP-1 cells.

Antibodies to ADT1-4-53 or antibodies derived from ADT1-4-53 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage identity) may have K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM) and/or K _D to cynomolgus monkey TRDV1 of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

The antibody to ADT1-4-173 or the antibody derived from ADT1-4-173 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

Antibodies to ADT1-4-2 or antibodies derived from ADT1-4-2 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage of identity) may have K _D to human TRDV of less than about 1nM (preferably less than about 0.5 nM) and/or K _D to cynomolgus monkey TRDV1 of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells

Antibodies to ADT1-4-8 or antibodies derived from ADT1-4-8 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-82 or the antibody derived from ADT1-4-82 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-83 or the antibody derived from ADT1-4-83 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

Antibodies to ADT1-4-3 or antibodies derived from ADT1-4-3 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-84 or the antibody derived from ADT1-4-84 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-86 or the antibody derived from ADT1-4-86 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

The antibody to ADT1-4-95 or the antibody derived from ADT1-4-95 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) may have a K _D to human TRDV of less than about 10nM (preferably less than about 1 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 50 nM.

The antibody to ADT1-4-1 or the antibody derived from ADT1-4-1 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV of less than about 1nM (preferably less than about 0.5 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells.

Antibodies to ADT1-4-6 or antibodies derived from ADT1-4-6 (e.g., fragments thereof, variants having one or more amino acid substitutions relative thereto, or having a certain percentage identity) may have K _D to human TRDV nM (preferably less than about 5 nM) and/or K _D to cynomolgus monkey TRDV1 of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 1nM (preferably less than about 0.5 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 10nM (preferably less than about 1 nM) for killing THP-1 cells.

The antibody to ADT1-4-138 or the antibody derived from ADT1-4-138 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a percentage of identity) may have a K _D to human TRDV1 of less than about 100nM (preferably less than about 50 nM) and/or a K _D to cynomolgus monkey TRDV1 of less than about 100nM (preferably less than about 50 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 50nM (preferably less than about 10 nM) and/or an IC90 for TCR downregulation of less than about 100nM (preferably less than about 50 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 10nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 50nM (preferably less than about 10 nM) for killing THP-1 cells.

Pharmacologically active antibodies to ADT1-7 lineages

The antibody to ADT1-7-10 or the antibody derived from ADT1-7-10 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-15 or the antibody derived from ADT1-7-15 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-17 or the antibody derived from ADT1-7-17 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-18 or the antibody derived from ADT1-7-18 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-19 or the antibody derived from ADT1-7-19 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-20 or the antibody derived from ADT1-7-20 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 5nM (preferably less than about 1 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 5 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 5nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells.

The antibody to ADT1-7-22 or the antibody derived from ADT1-7-22 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage of identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-23 or the antibody derived from ADT1-7-23 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-42 or the antibody derived from ADT1-7-42 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV of less than about 10nM (preferably less than about 5 nM).

The antibody to ADT1-7-3 or the antibody derived from ADT1-7-3 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR downregulation of less than about 5nM (preferably less than about 1 nM) and/or an IC90 for TCR downregulation of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 5nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 10nM (preferably less than about 5 nM) for killing THP-1 cells.

The antibody to ADT1-7-61 or the antibody derived from ADT1-7-61 (e.g., a fragment thereof, a variant having one or more amino acid substitutions relative thereto, or a certain percentage identity) can have a K _D to human TRDV1 of less than about 10nM (preferably less than about 1 nM). Alternatively or additionally, such antibodies may have an IC50 for TCR down-regulation of less than about 50nM (preferably less than about 10 nM). Alternatively or additionally, such antibodies may have an IC50 of less than about 5nM (preferably less than about 1 nM) for killing THP-1 cells and/or an IC90 of less than about 50 for killing THP-1 cells.

Other functional Properties of antibodies

The antibodies of the invention have advantageous functional characteristics. In particular, unlike prior art anti-vδ1 antibodies that focus on vδ1T cell depletion, the antibodies of the present invention may be used to activate vδ1T cells. Although they cause down-regulation of the TCR on the T cells to which they bind, they do not lead to depletion of vδ1T cells, but rather they stimulate T cells and are therefore useful in therapeutic settings that would benefit from activation of this T cell compartment. Activation of vδ1T cells was evident by TCR down-regulation, changes in activation markers such as CD25 and Ki67 and degranulation marker CD107 a. Activation of vδ1T cells in turn leads to release of inflammatory cytokines such as infγ and tnfα, thereby promoting immune licensing. Surprisingly, antibodies with suitably high affinity for TRDV1 cause increased killing of vδ T cells, and unlike antibodies that target CD3, for example, can provide high affinity antibodies without the adverse effects associated with large scale activation by CD 3. In turn, high affinity antibodies can induce strong immunostimulation by Tumor Infiltrating Lymphocytes (TIL). This can be achieved by minimal depletion or killing of vδ1 cells. Thus, the antibodies of the invention may be considered agonistic antibodies.

In one embodiment of the invention, there is provided an anti-vδ1 antibody or antigen binding fragment thereof, said anti-vδ1 antibody or antigen binding fragment thereof being characterized in that it:

a) Causing TCR down on vδ1T cells;

b) Does not exhibit CDC or ADCC; and

C) The vδ1T cells were not depleted.

In some embodiments, the anti-vδ1 antibody or antigen binding fragment further stimulates vδ1T cells proliferation.

An antibody or antigen-binding fragment thereof may be further defined as binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 10nM, preferably less than about 5 nM.

The antibody or antigen binding fragment thereof may be further defined as having advantageous K _D, IC50 and/or IC90 values as described above.

T cell depletion is the process of T cell death removal or reduction. Reference to an antibody or antigen binding fragment that does not deplete vδ1T cells refers to a population of viable vδ1T + cells that is less than about 30% or less than about 20% (preferably less than about 10%) depleted when incubated by one or more antibodies of the present invention as described herein (e.g. when the antibodies are provided as IgG1 antibodies) and as measured in a controlled study (e.g. via a controlled flow cytometry method or via other established controlled assays, as described in fig. 41 and 55) by any suitable method.

ADCC and CDC are possible mechanisms of occurrence of T cell depletion. References herein to antibodies or antigen binding fragments that do not result in ADCC or CDC refer to a population of viable vδ T + cells that is less than about 30% or less than about 20% (preferably less than about 10%) depleted when incubated by one or more antibodies of the present invention as described herein (e.g., when the antibodies are provided as IgG1 antibodies) as measured by any suitable method (e.g., via a controlled flow cytometry method or via other established controlled assays, as described in fig. 41).

In one embodiment, an anti-vδ1 antibody or antigen-binding fragment thereof is provided, said anti-vδ1 antibody or antigen-binding fragment thereof being characterized in that it does not induce secretion of IL-17A. IL-17A (interleukin-17A) is a pro-tumor cytokine produced by activated T cells. IL-17A can enhance tumor growth and inhibit anti-cancer immune responses. As shown in fig. 63G to I, when added to a cell population comprising human lymphocytes including vδ1+ cells, the anti-vδ1 antibody did not induce secretion of IL-17A, whereas the comparative anti-CD 3 antibody such as OKT3 induced IL17A in this case. Thus, references herein to an antibody or antigen-binding fragment that does not induce secretion of IL-17A refer to an antibody that induces less than about 30%, or less than about 20%, or less than about 10% of the IL-17A secretion induced by a comparison anti-CD 3 antibody in this case (represented by OKT 3; anti-CD 3 comparison antibody used in FIGS. 63G-I).

Antibody modification

Antibodies and fragments thereof may be modified in other ways using known methods. Sequence modifications to the antibody molecules described herein can be readily incorporated by those skilled in the art. The following examples are non-limiting.

During the discovery of antibodies and recovery sequences from phage libraries, the desired antibody variable domains can be reformatted into full-length IgG by subcloning. To accelerate this process, restriction enzymes are often used to transfer the variable domains. These unique restriction sites may introduce additional/alternative amino acids and be located away from canonical sequences (such canonical sequences may be found in, for example, the International ImMunoGeneTics [ IMGT ] information system, see http:// www.imgt.org). These may be introduced as kappa or lambda light chain sequence modifications.

Light chain modification

The light chain variable sequences can be cloned during reformatting into full length IgG using restriction sites (e.g., nhe1-Not 1). More specifically, at the N-terminus of the light chain, additional Ala-Ser sequences were introduced in the parent (unaffinity matured) antibody to support cloning. Preferably, this additional AS sequence is then removed during further development to generate a canonical N-terminal sequence. Thus, in one embodiment, the light chain-containing antibodies described herein do not contain AS sequences at their N-terminus, i.e., SEQ ID NO: 26. 118, 282 through 290 or 313 do not contain an initial AS sequence. The N-terminus of the light chain sequence of the affinity matured antibody does not already contain this AS motif.

Additional amino acid changes may be made to support cloning. For example, for the parent antibodies described herein (i.e., B07, C05, E04, F07, G06, G09, B09, G10, G04, and E07) with kappa light chains, valine to alanine changes were introduced at kappa light chain variable domain/constant domain boundaries to support cloning when preparing full length sequences. This resulted in kappa constant domain modification. Specifically, this resulted in a constant domain starting with RTAAAPS (from the NotI restriction site). Preferably, the sequence may be modified during further development to generate a canonical kappa light chain constant region starting at RTVAAPS. Such modifications do not alter the functional properties of the antibody. Thus, in some embodiments, the kappa light chain-containing antibodies described herein contain a constant domain (e.g., as in SEQ ID NO: 296) that begins with the sequence RTV.

As another example, for the antibodies described herein (particularly E01 and C08), a lysine to alanine sequence change was introduced at the lambda light chain variable domain/constant domain boundary to support cloning. This results in lambda constant domain modification. Specifically, this resulted in a constant domain starting with GQPAAAPS (from the NotI restriction site). Preferably, the sequence may be modified during further development to generate a canonical lambda light chain constant region starting at GQPKAAPS. Thus, in some embodiments, the lambda light chain-containing antibodies described herein contain a constant domain that begins with sequence GQPK.

Heavy chain modification

Typically, the human variable heavy chain sequence begins with either basic glutamine (Q) or acidic glutamic acid (E). However, both sequences are known to be converted to the acidic amino acid residue pyroglutamic acid (pE). The conversion of Q to pE resulted in a change in the charge of the antibody, whereas the conversion of E to pE did not change the charge of the antibody. Thus, to avoid variable charge changes over time, one option is to first modify the starting heavy chain sequence from Q to E. Thus, in one embodiment, the heavy chain of an antibody described herein having a Q residue at the N-terminus of the heavy chain may contain a Q to E modification at the N-terminus. Specifically, SEQ ID NO: 1. the starting residue of any one of 106, 276 to 279 or 312 may be modified from Q to E. It will be appreciated that this embodiment is also applicable to any embodiment in which these sequences are incorporated into, for example, a full length antibody or antigen binding fragment thereof. In some embodiments, it may be advantageous to replace the E residue at the N-terminus of the heavy chain with an E residue. Thus, in some embodiments, SEQ ID NO: the E residue at the N-terminus of any one of 2 to 25, 107 to 117, 273 to 275, 280 or 281 may be substituted with a Q residue.

Furthermore, the C-terminus of the IgG1 constant domain ends with PGK. However, terminal basic lysines (K, EU position 447) are typically cleaved during expression (e.g., in CHO cells). This in turn leads to a change in charge of the antibody by a different loss of C-terminal lysine residues. Thus, one option is to first remove lysine, thereby producing a uniform and consistent heavy chain C-terminal sequence ending with PG. Another option is to also remove end G (EU position 446). Thus, in one embodiment, the terminal K or terminal GK of the heavy chain of an antibody described herein is removed from the C-terminus of the heavy chain.

In some embodiments, the antibody or antigen binding fragment thereof contains a modified effector function by altering the saccharide attached to Asn 297 (EU numbering scheme). In another such modification, asn 297 is not fucosylated or exhibits reduced fucosylation (i.e., a defucosylated antibody or a nonfucosylated antibody). Fucosylation involves the addition of the sugar fucose to molecules, such as fucose linkages to N-glycans, O-glycans and glycolipids. Thus, in defucosylated antibodies, fucose is not attached to the carbohydrate chain of the constant region. Antibodies may be modified to prevent or inhibit fucosylation of the antibodies. Typically, glycosylation modification involves expression of the antibody or antigen-binding fragment thereof in a host cell containing alternative glycosylation processing capacity by targeted engineering or by targeted or occasional host or clonal selection (see, e.g., example 13). These and other effector modifications are further discussed in recent reviews, such as Xinhua Wang et al (2018) Protein & Cell 9:63-73 and Pereira et al (2018) mAbs 10 (5): 693-711 and are incorporated herein.

Optional allotypic modification

During antibody discovery, specific human allotypes may be employed. Optionally, the antibodies may be converted to different human allotypes during development. As a non-limiting example, for kappa chains, there are three human allotypes, termed Km1,2 and Km3, which define three Km alleles (using allotypic numbering): km1 is associated with valine 153 (IMGT V45.1) and leucine 191 (IMGT L101); km1,2 is associated with alanine 153 (IMGT a 45.1) and leucine 191 (IMGT L101); and Km3 is associated with alanine 153 (IMGT a 45.1) and valine 191 (IMGT V101). Optionally, the sequence may thus be modified from one allotype to another by standard cloning methods. For example, an L191V (IMGT L101V) change will convert Km1,2 allotypes to Km3 allotypes. For further reference to such allotypes, see Jefferis and Lefranc (2009) MAbs 1 (4): 332-8, which are incorporated herein by reference.

Thus, in one embodiment, the antibodies described herein contain amino acid substitutions derived from another human allotype of the same gene. In another embodiment, the antibody contains an L191V (IMGT L101V) substitution to the kappa chain to convert the c-domain from km1,2 to km3 allotypes.

Multispecific antibodies

Antibodies of the invention may be monospecific. In some embodiments, the antibody is not a multispecific antibody. In some embodiments, the antibody is not a bispecific antibody. However, the invention also provides multispecific antibodies. Thus, antibodies can bind additional targets and are therefore bispecific or multispecific. A multispecific antibody may be specific for different epitopes of one target polypeptide, or may be specific for more than one target polypeptide. Thus, in one embodiment, the antibody or antigen binding fragment thereof comprises a first binding specificity for vδ1 and a second binding specificity for a second target epitope or antigen.

In particular, the invention provides a novel class of multispecific antibodies that target the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) and a second antigen. The second antigen may be, for example, a cancer antigen or a cancer-associated antigen (e.g., TAA), and thus the antibody may be referred to as a T cell cement (TCE). Alternatively, the second antigen may be, for example, an immunomodulatory antigen, and thus the antibody may be a dual immunomodulatory antibody. The multispecific antibodies of the present invention can thus be divided into two classes. The first class is multispecific antibodies as T cell cements. The second class is multispecific antibodies that are dual immunomodulators (Dis).

The TCE of the present invention provides several advantages over prior art TCEs. In particular, TCEs can overcome many of the challenges associated with TCEs of the prior art by targeting T cell receptor complexes via entirely new discrete mechanisms. Indeed, by specifically targeting (and activating) the T cell receptor complex via only the epitope bound to the TRDV1 domain, a number of advantages can be achieved, including:

Engaging only a subset of T cells, but not all T cells (e.g., engaging T-reg may be undesirable in a cancer setting);

only the junction is predominantly 'tissue resident' and there is a subset of T cells (TRDV 1 +t cells) that are normally positively correlated with a good prognosis in the cancer/tumor environment;

Activation of the T cell receptor complex via TRDV a binding, thereby providing more selectivity (e.g., increasing the affinity of the binding domain, as in the antibodies of the invention). For example, by developing recombinant TCEs that bind to the T cell receptor complex only via the TRDV1 domain, rather than via CD3, increased affinity can drive more advantageous functions. For example, high affinity TRDV1 binding to TCE can activate but not deplete T cells; and/or

Joining the TCE complex via the novel method and via the recombinant TRDV binding domain may cause less detrimental effects, thereby reducing the need to attenuate Fc function in the TCE moiety. TCEs with unabated Fc function are generally expected to induce antibody dependent cell-mediated cytotoxicity (ADCC) to deplete antibody-recognized γδ T cell populations. However, also avoiding toxicity/safety complications by attenuating such functions, e.g. by engaging cd16+ or cd32+ or cd64+ immune cells, or by reducing the half-life of bispecific antibodies (e.g. if smaller bispecific antibody fragments such as BITE are used), may also attenuate the potential important efficacy angle. Methods of reducing fcγr and TCE interactions (e.g., using IgG formats intended to reduce the interactions) are expected to reduce Fc-mediated TCE immobilization and reduce TCR aggregation by cross-linking with the immobilized TCE. Reducing the need to attenuate Fc function in the TCE moiety may provide additional selectivity, for example, by allowing TRDV1 TCE to bind to TRDV1+ cells via one binding domain, to bind to a second cell type (e.g., cancer cells) via a second binding arm, and to bind to other effector cells, such as cd16+ or cd32+ or cd64+ immune cells via a functional Fc domain.

Activating vδ1 TCR without depleting vδ1+ T cells. Thus, the antibodies described herein can bind to and activate the CD3/γδ TCR complex and confer surface expression down-regulation and loss thereto without unnecessary depletion of vδ1T cells (see figure 55). This enables the antibodies described herein to be used as medicaments for treating a disease or disorder, thereby ameliorating at least one sign or symptom of the disease or disorder by a mechanism involving activation of blood, tissue, and tumor-resident vδ1+ T cells.

Thus, through discovery as described herein, the inventors have generated a new class of recombinant TCEs. In particular, the inventors have discovered a new class of TCEs that bind T cell receptors via the TRDV1 domain, but not the other domains in the T cell receptor signaling complex. More specifically, the present inventors have discovered a new class of TCEs that bind to the complex via an activating epitope on TRDV a and can bind with higher affinity without potentially conferring some of the previously reported deleterious effects associated with binding to high affinity T cell receptor complexes. Furthermore, such novel TCEs may be conjugated in a manner that may allow wild-type Fc function, thereby also providing additional potency potential.

The DI of the present invention provides a completely unique and novel approach to dual immunomodulatory target engagement, providing a vast range of possible novel therapies requiring immunomodulation, such as cancer. The DI platform of the present invention provides a new class of therapeutic approaches that can provide an important replacement or retrofit to existing DI approaches. It was previously not contemplated that TRDV1 specific binding functions as described herein could be incorporated into the DI format.

The multispecific antibodies of the invention may also exhibit improved properties compared to equivalent monospecific antibodies. For example, the multispecific antibodies of the invention may also exhibit improved properties compared to monospecific antibodies having the same antigen-binding domains as the components of the multispecific antibodies. In some embodiments, for example, the recombinant multispecific antibody confers increased γδ T cell mediated cytotoxicity to diseased cells expressing the second epitope compared to the cytotoxicity conferred by an equivalent amount of the first monospecific antibody. The multispecific antibodies of the present invention may also exhibit increased cytotoxicity to diseased cells while still retaining healthy cells.

The identity of the second antigen determines whether the antibody belongs to one of two categories as discussed herein: t cell cement (TCE) antibodies or Dual Immunomodulatory (DI) antibodies.

In embodiments related to TCE, the second antigen is a cancer antigen or a cancer-related antigen. In such embodiments, the antibody specifically binds to a first target epitope, wherein the first target epitope is an epitope of a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR); and a second target epitope, wherein the second target epitope is an epitope of a cancer antigen or a cancer-associated antigen. The identity of a particular possible second antigen in this class will be discussed herein. However, the second antigen may be any antigen expressed by a cancer cell that promotes vδ1-T cell-mediated killing of the cancer cell (e.g. direct killing or via an immunopermissive effect that signals other immune cells upon tumor cell binding). Such vδ1 cell-mediated killing of cancer cells is facilitated by co-locating the vδ1-T cells and the cancer cells, and activating the vδ1-T cells via binding of multispecific antibodies (in particular binding to activating epitopes of the vδ1-T cells). The present invention exemplifies a completely new platform for TCE type antibodies.

In some embodiments, the second antigen is not an antigen of ovarian cancer. In some embodiments, the second antigen is not an antigen of mov19+ ovarian cancer. In some embodiments, the multispecific (suitably bispecific) antibody does not specifically bind to a mov19+ ovarian cancer cell. In some embodiments, the multispecific (suitably bispecific) antibody does not specifically bind to the α -folate receptor (α -FR). alpha-FR is also known as folate receptor 1, FOLR1, folate receptor alpha or fra. It is encoded by FOLR1 gene (UniProt accession number P15328) and has the sequence of SEQ ID NO:390. in some embodiments, the multispecific (suitably bispecific) antibody does not specifically bind to an epitope to which scFv MOV19 binds.

In some embodiments, the antibody or antigen-binding fragment thereof is a bispecific antibody, wherein the second antigen is not an alpha-folate receptor.

In some embodiments, the multispecific antibody is a human recombinant antibody encoded by one or more recombinant nucleic acid open reading frames expressed by a recombinant host cell. In some embodiments, the multispecific antibody is not a rodent or other non-human antibody derived from a B cell fusion hybridoma technique. In some embodiments, the multispecific antibody does not comprise non-human IgG constant domain sequences found only in non-human animal species, such as those found in a rodent-derived hybridoma.

In embodiments related to DI, the second antigen is an immunomodulatory antigen. In such embodiments, the antibody specifically binds to a first target epitope, wherein the first target epitope is an epitope of a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR); and a second target epitope, wherein the second target epitope is an immunomodulatory antigen. An "immunomodulatory antigen" is an antigen that modulates (e.g., promotes) antibody-mediated and/or cell-mediated immunity. An immunomodulatory antigen is an antigen that is present on the cell surface of T cells. In embodiments of the invention in which the second antigen is an immunomodulatory antigen, the second antigen is not a component of a T cell receptor or T cell receptor complex. For example, in embodiments of the invention in which the second epitope is an epitope of an immunomodulatory antigen, the second epitope is not an epitope of TRDV a. In a preferred embodiment of the invention wherein the second epitope is an epitope of an immunomodulatory antigen, the second epitope is not an epitope of a T cell receptor complex. For example, in some embodiments, the second epitope is not an epitope of CD 3. Thus, the antibodies of these embodiments are "dual immunomodulators" in that they specifically bind to T cells via TRDV a1 and can additionally bind to T cells via a second, different epitope, wherein the epitope is not an epitope of the T cell receptor complex. Exemplary second antigens include, for example, immune checkpoint inhibitors PD-L1, PD-1, OX40, CTLA-4, LAG-3, TIM-3, TIGIT, and VISTA. For example, solid tumors recruit immunosuppressive cells, such as bone marrow-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), and regulatory T cells (tregs), all of which suppress the activity of cytotoxic T cells. Thus, the most efficient use of DI in solid tumors may require the use of multi-specific moieties in combination to target T cell regulatory pathways to help overcome immunosuppressive TMEs and convert immune rejection or immune desert "cold" tumors to inflammatory "hot" tumors. However, the present invention is not limited to a particular second immunomodulatory antigen, as it provides a completely new platform for DI-type antibodies.

In a preferred embodiment, the multispecific antibodies (suitably bispecific antibodies) of the invention do not specifically bind (or directly interact with) CD3. In a preferred embodiment, the second antigen is not CD3.

In some embodiments, the second antigen is not CD3 or alpha-folate receptor.

In some embodiments, the second target epitope is on an antigen expressed by vδ1+ T cells (particularly an antigen expressed on the surface of vδ1+ T cells). In other embodiments, the second target epitope is on an antigen expressed by another cell (i.e., a cell that is not a vδ1+ T cell) (particularly an antigen expressed on the surface of a cell that is not a vδ1+ T cell). For example, the second antigen may be expressed by a cancer cell.

References herein to an antigen "on" a cell refer to an antigen expressed on or associated with (outside of) the cell surface membrane of such a cell.

In various embodiments, the second target epitope is an epitope of a cancer antigen or a cancer-associated antigen. In various embodiments, the cancer antigen or cancer-associated antigen is one selected from the group consisting of: AFP, AKAP-4, ALK, alpha fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCA, bcr-abl, beta-catenin, beta-HCG, beta-human chorionic gonadotrophin, BORIS, BTAA, CA, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, CAM 17.1, CAM43, carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33/IL3Ra, CD68\P1, CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG 4), C-Met, CO-029, CSPG4, cyclin B1, cyclophilin C-related protein, CYP1B1, E2A-PRL, EGFR, EGFRvIII, ELF2M, epCAM, ephA, ephran B2, ephran-Bar virus antigen EBVA, ERG (TMPRSS 2ETS fusion gene) ETV6-AML, FAP, FGF-5, fos associated antigen 1, fucosyl GM1, G250, ga733\EpCAM, GAGE-1, GAGE-2, GD3, glioma associated antigen, globoH, glycolipid F77, GM3, GP100 (Pmel 17), H4-RET, HER-2/Neu/ErbB-2, high molecular weight melanoma associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin Growth Factor (IGF) -I, intestinal carboxylesterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, legumain, LMP2, M344, MA-50, HIV-2, mac-2 binding protein 、MAD-CT-1、MAD-CT-2、MAGE、MAGE A1、MAGE A3、MAGE-1、MAGE-3、MAGE-4、MAGE-5、MAGE-6、MART-1、MART-1/MelanA、M-CSF、 melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin, MG7-Ag, ML-IAP, MN-CA IX, MOV18, MUC1, mum-1, hsp70-2, MYCN, MYL-RAR, NA17, NB/70K, neuronal-glial antigen 2 (NG 2), neutrophil elastase, nm-23H1, nuMa, NY-BR-1, NY-CO-1, NY-ESO-1, OY-TES1, p15, p16, p180erbB3, p185erbB2, p53 mutant, page4, PAX3, PAX5, PDGFR-beta, PLAC1, polysialic acid, POLYSAC prostate cancer tumor antigen-1 (PCTA-1), prostate specific antigen, prostatic Acid Phosphatase (PAP), protease 3 (PR 1), PSA, PSCA, PSMA, RAGE-1, ras mutant, RCAS1, RGS5, rhoC, ROR1, RU2 (AS), SART3, SDCCAG, sLe (a), sperm protein 17, SSX2, STn, survivin, TA-90, TAAL, TAG-72, telomerase, thyroglobulin, tie 2, TLP, tn, TPS, TRP-1, TRP-2, TSP-180, tyrosinase, VEGF, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72.

In some embodiments, the second target epitope is in the ErbB subfamily. Erb-B1 (EGFR), erb-B2 (HER 2) are members of the Receptor Tyrosine Kinase (RTK) superfamily subclass I. In some embodiments, the second target epitope is an RTK. RTKs share a similar protein structure, comprising an extracellular ligand binding domain and a single transmembrane helix. They are then further subdivided into independent subclasses primarily according to their intracellular Tyrosine Kinase Domain (TKD) and carboxyl (C-) terminal tail properties. The extracellular domain region of RTKs exhibits a variety of conserved elements, including immunoglobulin (Ig) -like or Epidermal Growth Factor (EGF) -like domains. In some embodiments, the second epitope is an epitope of a receptor tyrosine kinase. Examples of RTK families include VEGFR2, EGFR, c-MET, IGF-I receptor, PDGFR- β, CD115, CD117, CD140A, CD, 140, B, CD167a, CD167b, CD172g, CD220, CD246, CD303, CD331, CD332, CD333, and CD340.

For example, in some embodiments, the second target epitope is HER2 (human epidermal growth factor receptor 2). HER2 (also known as ErbB-2 or CD 340) is a cancer-associated antigen and is an example of a receptor tyrosine kinase in the ErbB subfamily. HER2 expression is lower in healthy tissues and increased up to 40-100 fold in her2+ cancers compared to normal tissues. Her2 overexpression is associated with many breast, gastric, esophageal, ovarian, endometrial, NSCLC and colorectal cancers. In many cancers, HER2 dimerizes with other ErbB receptors and leads to activation of various downstream signaling pathways, which in turn lead to uncontrolled proliferation and resistance to apoptosis. Overexpression of HER2 is associated with lower survival, so it is a target for improved prognosis, as well as a tumor marker. Monoclonal antibodies that specifically bind to epitopes of HER2 are well known in the art. Trastuzumab, for example, is a monoclonal antibody that specifically binds to an epitope of HER 2.

In some embodiments, the second target epitope is EGFR. EGFR (epidermal growth factor receptor) is a cancer-associated antigen and is an example of a receptor tyrosine kinase in the ErbB subfamily. EGFR is expressed in multiple organs and plays an important role in initiating signaling that directs the behavior of epithelial cells and tumors of epithelial origin. EGFR-mediated signaling is also involved in controlling cell proliferation, migration, survival and metastasis by modulating different cellular pathways. Like other receptor tyrosine kinases, mutations that affect EGFR activity or cause EGFR upregulation are associated with many cancers. Indeed, genetic alterations of EGFR are observed in up to 30% of solid tumors, and are often associated with poor prognosis. EGFR-expressing tumor growth may be restricted by inhibiting EGF binding to extracellular domains or by disrupting EGFR signaling by inhibiting intracellular tyrosine kinase activity. Thus, EGFR inhibitors may be anticancer agents. In fact, some tumor cells rely on EGFR signaling and therefore have "oncogene addiction", which makes this receptor an attractive therapeutic target. Monoclonal antibodies that specifically bind to epitopes of EGFR are well known in the art. For example, cetuximab is a monoclonal antibody that specifically binds to an epitope of EGFR.

In some embodiments, the second target epitope is an epitope of a B lymphocyte antigen. Examples of antigens that are significantly expressed on B cells include CD1d、CD5、CD10、CD11b、CD19、CD20、CD21、CD22、CD23、CD24、CD32A、CD32B CD37、CD39、CD40、CD45、CD52、CD72、CD79a、CD79b、CD138、CD166、CD179A、CD179B、CD180、CD185、CD150、CD213a1、CD213a2、CD217、CD244、CD255、CD229、CD232、CD267、CD268、CD269、CD274、CD277、CD279、CD290、CD300A、CD300C、CD305、CD307a、CD307b、CD307c、CD307d、CD307e、CD316、CD319、CD327、CD352 and CD361. For example, in some embodiments, the second target epitope is CD19.CD19 (cluster of differentiation 19) is a cancer-associated antigen. CD19 is also an example of a type I transmembrane glycoprotein in the immunoglobulin superfamily. In some embodiments, the second target is an epitope present on a member of the immunoglobulin superfamily (IgSF). Examples of this family include CD2、CD3、CD4、CD7、CD8、CD19、CD79A、CD79B、CD28、CD48、CD58、CD80、CD86、CD90、CD96、CD147、CD150、CD155、CD229、CD244、CD273、CD274、CD276.CD19 being widely expressed on B cells throughout their development, the surface density of CD19 increasing as B cells mature. CD19 is involved in the recruitment of signaling proteins from the cytoplasm. CD19 is also involved in B cell receptor signaling pathways and is critical to the function of B cell receptors. Its expression on B cells makes it a useful target against leukemia and neoplastic lymphocytes, as well as a diagnostic biomarker for B cell-induced cancers. CD19 mutations can lead to reduced antibody production and immunodeficiency, and thus CD19 can also be a target for the treatment of autoimmune diseases. Monoclonal antibodies that specifically bind to an epitope of CD19 are well known in the art. For example, bleb mab is a monoclonal antibody that specifically binds to an epitope of CD19.

In some embodiments, the second target epitope is present on a tumor stroma antigen. Examples of tumor stromal antigens include fapα, CD29, CD44, CD73, CD105, and CD166. For example, in some embodiments, the second target epitope is fapα (fibroblast activation protein α). Fapα is a cancer-associated antigen, also known as serine membrane protease (seprase) or prolyl endopeptidase FAP. It is selectively expressed in the stroma of a range of epithelial cancers. Fapα is an example of a cell surface serine protease in the dipeptidyl peptidase family. Fapα is expressed by cancer-associated fibroblasts (CAF) that play an important role in the tumor microenvironment. Other molecules that are selectively expressed on CAF include CD10, CD90, CD140A and CD140B. In some embodiments, the second epitope is an epitope present on a molecule that is selectively expressed on CAF. More than 90% of the epithelial cancers (breast, CRC skin and pancreatic cancers) were found to express fapα on CAF surfaces in the surrounding stroma. CAF secretes the chemokine CXCL12, which binds to CXCR4 on T cells and has an immunosuppressive effect. Fapα was found to be expressed in invasive melanoma cell lines and increased significantly in patients with poor breast cancer outcome and survival. Fapα has collagenase and dipeptidase activities and promotes tumor growth, migration, invasion, metastasis, and ECM degradation. Normal healthy adult tissue has no detectable fapα expression outside of the tissue remodelling or wound healing areas, so fapα is a promising anticancer target because it is expressed almost exclusively in tumor stroma and fapα plays a direct role in various aspects of cancer progression. Monoclonal antibodies that specifically bind to epitopes of fapα are well known in the art. For example, cetrimab is a monoclonal antibody that specifically binds to an epitope of fapα.

In some embodiments, the bispecific or multispecific antibody comprises a fapα binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:458, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:459 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:460, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:461, comprising the amino acid sequence of SEQ ID NO:462 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 463. In some embodiments, the bispecific or multispecific antibody comprises a fapα binding component comprising a nucleotide sequence comprising SEQ ID NO:456 or a VH consisting of said amino acid sequence and a nucleotide sequence comprising SEQ ID NO:457 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to fapα and comprises SEQ ID NO:402.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, a bispecific or multispecific antibody specifically binds to TRDV and fapα and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-A fapα binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:458, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:459 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:460, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:461, comprising the amino acid sequence of SEQ ID NO:462 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 463.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV a and fapα and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A fapα binding component comprising a nucleotide sequence comprising SEQ ID NO:456 or a VH consisting of said amino acid sequence and a nucleotide sequence comprising SEQ ID NO:457 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and FAPa and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A fapα binding component comprising the amino acid sequence of SEQ ID NO:402.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and fapα and comprises SEQ ID NO:414 and SEQ ID NO:415. in some embodiments, the antibody specifically binds to TRDV and fapα and comprises SEQ ID NO:504 and SEQ ID NO:415. in some embodiments, the antibody specifically binds to TRDV and fapα and comprises SEQ ID NO:505 and SEQ ID NO:415.

In some embodiments, the second target epitope is present on a cell surface glycoprotein. Protein glycosylation is an important and common post-translational modification. It is believed that more than 50% of human proteins are glycosylated to regulate protein function. Aberrant glycosylation is associated with a variety of diseases such as inflammatory skin diseases, diabetes, cardiovascular disorders, rheumatoid arthritis, alzheimer's disease and prion diseases, and cancer. Examples of cell surface glycoproteins include CD1a、CD1b、CD1c、CD1d、CD1e、CD3d、CD3e、CD3g、CD8a、CD8b、CD11a、CD21、CD36、CD42a、CD42b、CD42c、CD42d、CD43、CD66a、CD66f、CD177、CD235a、CD235b、CD236、CD238、CD243、CD227 and CD301. For example, in some embodiments, the second target epitope is Mesothelin (MSLN). MSLN is a cancer-associated antigen and is an example of a cell surface glycoprotein. MSLN is expressed in healthy cells (pleural, peritoneal and pericardial mesothelial cells) but is also expressed in a variety of cancers (malignant mesothelioma and pancreatic, cholangiocarcinoma, ovarian and lung adenocarcinoma, malignant mesothelioma, pancreatic, ovarian, endometrial, cholangiocarcinoma, gastric and pediatric acute myelogenous leukemia). MSLN is an example of a tumor differentiation antigen. The physiological function of MSLN is not yet clear, but MSLN is a useful target for therapy localization to msln+ tumors, or can be used as a tumor marker. Monoclonal antibodies that specifically bind to epitopes of MSLN are well known in the art. For example, anetuzumab (anetumab) is a monoclonal antibody that specifically binds to an epitope of MSLN.

In some embodiments, the bispecific or multispecific antibody comprises a MSLN binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:466, comprising the amino acid sequence of SEQ ID NO:467 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: VHCDR3 of the amino acid sequence of 468; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:469, comprising the amino acid sequence of SEQ ID NO:470 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:471, and VLCDR3 of the amino acid sequence of 471. In some embodiments, the bispecific or multispecific antibody comprises a MSLN binding component comprising a polypeptide comprising SEQ ID NO:464 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:465 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to MSLN and comprises the amino acid sequence of SEQ ID NO:404.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, a bispecific or multispecific antibody specifically binds to TRDV and MSLN and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-A MSLN binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:466, comprising the amino acid sequence of SEQ ID NO:467 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: VHCDR3 of the amino acid sequence of 468; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:469, comprising the amino acid sequence of SEQ ID NO:470 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:471, and VLCDR3 of the amino acid sequence of 471.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and MSLN and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A MSLN binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:464 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:465 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and MSLN and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A MSLN binding component comprising the amino acid sequence of SEQ ID NO:404

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and MSLN and comprises SEQ ID NO:414 and SEQ ID NO:416. in some embodiments, the antibody specifically binds to TRDV and MSLN and comprises SEQ ID NO:504 and SEQ ID NO:416. in some embodiments, the antibody specifically binds to TRDV and MSLN and comprises SEQ ID NO:505 and SEQ ID NO:416.

In various embodiments, the second target epitope is an epitope of an immunomodulatory antigen. An immunomodulatory antigen is an antigen that modulates (activates or represses) the immune system. In some embodiments, the immunomodulatory antigen is a cell surface protein (i.e., an antigen expressed on the surface of a cell, particularly an antigen expressed on the surface of an immune cell such as a lymphocyte, neutrophil, monocyte, or macrophage). The immunomodulator antigen may be expressed on vδ1+ T cells or may be expressed by different cells, for example cd4+ cells, cd8+ cells or different immune cells. The immunomodulatory antigen may be selected from the group ：B7-1(CD80)、B7-2(CD86)、B7-DC(CD273)、B7-H1(CD274)、B7-H2(CD275)、B7-H3(CD276)、B7-H4(VTCN1)、B7-H5(VISTA)、BTLA(CD272)、4-1BB(CD137)、CD137L、CD24、CD27、CD28、CD38、CD40、CD40L(CD154)、CD54、CD59、CD70、CTLA4(CD152)、CXCL9、GITR(CD357)、HVEM(CD270)、ICAM-1(CD54)、ICOS(CD278)、LAG-3(CD223)、OX40(CD134)、OX40L(CD252)、PD-1(CD279)、PD-L1(CD274)、TIGIT、CD314、CD334、CD335、CD337 and TIM-3 (CD 366) consisting of.

In some embodiments, the second target epitope is an epitope of a stimulatory immune checkpoint molecule. For example, in some embodiments, the second target epitope is OX40 (CD 134). OX40 is an immunomodulator antigen and is an example of a TNFR superfamily (TNFRSF) member. In some embodiments, the second target epitope is an epitope present on a TNFRSF molecule. These proteins are a superfamily of cytokine receptors characterized by their ability to bind Tumor Necrosis Factor (TNF) via an extracellular cysteine-rich domain. Examples include CD18, CD27, CD30, CD40, CD95, CD120a, CD120b, CD134, CD137, CD265, CD268, CD269, CD270, CD271, CD357, and CD358. One such example OX40 (CD 134) is the late co-stimulatory immune checkpoint receptor expressed on cd4+ and cd8+ T cells. OX40 is expressed higher on cd4+ T cells and is not constitutively expressed on naive T cells, since OX40 expression depends on complete activation of T cells. When activated (e.g., by OX 40L), OX40 promotes cd4+/cd8+ T cell activation, survival and expansion of effector T cells and memory T cells, and inhibits Treg activity. This inhibits immune evasion of the tumor. Because OX40 is a stimulatory target, therapies targeting OX40 activate OX 40-expressing immune cells to stimulate an immune response (e.g., a cd48+ T cell response) against a tumor. Monoclonal antibodies that specifically bind to epitopes of OX40 are well known in the art. For example, pergolizumab (pogalizumab) is a monoclonal antibody that specifically binds to an epitope of OX 40.

In some embodiments, the bispecific or multispecific antibody comprises an OX40 binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:490, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:491 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:492, a VHCDR3 of an amino acid sequence; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:493, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:494 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 495. In some embodiments, the bispecific or multispecific antibody comprises an OX40 binding component comprising a polypeptide comprising SEQ ID NO:488 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:489 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to OX40 and comprises the amino acid sequence of SEQ ID NO:410.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and OX40 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-An OX40 binding component, said OX40 binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:490, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:491 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:492, a VHCDR3 of an amino acid sequence; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:493, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:494 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 495.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and OX40 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-An OX40 binding component comprising an amino acid sequence comprising SEQ ID NO:488 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:489 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and OX40 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-An OX40 binding component comprising the amino acid sequence of SEQ ID NO:410.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and OX40 and comprises SEQ ID NO:414 and SEQ ID NO:419. in some embodiments, the antibody specifically binds to TRDV and OX40 and comprises SEQ ID NO:504 and SEQ ID NO:419. in some embodiments, the antibody specifically binds to TRDV and OX40 and comprises SEQ ID NO:505 and SEQ ID NO:419.

In some embodiments, the second target epitope is an epitope present on a TNF superfamily member. For example, in some embodiments, the second target epitope is an epitope of 4-1BB (CD 137). 4-1BB is an immunomodulator antigen and is also an example of a member of the TNF superfamily. This is a superfamily of proteins containing type II transmembrane proteins that contain TNF homology domains, including CD70, CD137, CD153, CD154, CD252, CD253, CD254, CD256, CD257 and CD258.4-1BB (CD 137) is an inducible, co-stimulatory immune checkpoint receptor expressed on T cells as well as NK cells, dendritic Cells (DCs), monocytes, neutrophils and B cells. 4-1BB is a stimulatory antigen, and is specifically expressed on activated CD8+ T cells. In vitro 4-1BB stimulates the expansion of CD4+ T cells, CD8+ T cells, macrophages and DCs and the production of cytokines. In vivo 4-1BB is biased towards CD8+ T cell activation and exhibits potent anti-tumor activity, as cross-linking of 4-1BB has been shown to enhance T cell proliferation, IL-2 secretion, survival and cytolytic activity. Since 4-1BB is a stimulatory target, therapies targeting 4-1BB activate immune cells expressing 4-1BB to stimulate an immune response against a tumor (e.g., cytolytic CD8+ T cell response). Monoclonal antibodies that specifically bind to epitopes of 4-1BB are well known in the art. For example, wu Tuolu mab (utomilumab) is a monoclonal antibody that specifically binds to an epitope of 4-1 BB.

In some embodiments, the bispecific or multispecific antibody comprises a 4-1BB binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:482, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:483 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 484; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:485, comprising the amino acid sequence of SEQ ID NO:486 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:487, a VLCDR3 of the amino acid sequence. In some embodiments, the bispecific or multispecific antibody comprises a 4-1BB binding component comprising a polypeptide comprising SEQ ID NO:480 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:481 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to 4-1BB and comprises the amino acid sequence of SEQ ID NO:408.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, a bispecific or multispecific antibody specifically binds to TRDV1 and 4-1BB and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-A 4-1BB binding component, said 4-1BB binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:482, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:483 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 484; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:485, comprising the amino acid sequence of SEQ ID NO:486 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:487, a VLCDR3 of the amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV1 and 4-1BB and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A 4-1BB binding component comprising a polypeptide comprising SEQ ID NO:480 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:481 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV1 and 4-1BB and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A 4-1BB binding component, said 4-1BB binding component comprising the amino acid sequence of SEQ ID NO:408

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and 4-1BB and comprises SEQ ID NO:414 and SEQ ID NO:418. in some embodiments, the antibody specifically binds to TRDV and 4-1BB and comprises the amino acid sequence of SEQ ID NO:504 and SEQ ID NO:418. in some embodiments, the antibody specifically binds to TRDV and 4-1BB and comprises the amino acid sequence of SEQ ID NO:505 and SEQ ID NO:418.

In some embodiments, the second target epitope is an immune checkpoint inhibitor molecule. For example, in some embodiments, the second target epitope is TIGIT (T cell immune receptor with Ig and ITIM domains). TIGIT is an immunomodulator antigen and is an example of an immune checkpoint inhibitor expressed on T cells (including γδ T cells and NK cells). TIGIT plays a role in immune homeostasis and in the prevention of autoimmunity by binding to its ligand (PVR/CD 155) leading to T cell inhibition. TIGIT is overexpressed on tumor-infiltrating lymphocytes. Therapeutic blockade of TIGIT is desirable because it increases T cell proliferation, cytokine production, and degranulation. Monoclonal antibodies that specifically bind to epitopes of TIGIT are well known in the art. For example, the tirizidine Li Youshan antibody is a monoclonal antibody that specifically binds to an epitope of TIGIT.

In some embodiments, the bispecific or multispecific antibody comprises a TIGIT binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:498, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:499 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:500, a VHCDR3 of an amino acid sequence; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:501, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:502 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:503, and a VLCDR3 of the amino acid sequence of seq id no. In some embodiments, the bispecific or multispecific antibody comprises a TIGIT binding component comprising a polypeptide comprising SEQ ID NO:496 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:497 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to TIGIT and comprises the amino acid sequence of SEQ ID NO:412.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, a bispecific or multispecific antibody specifically binds to TRDV and TIGIT and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-A TIGIT binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:498, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:499 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:500, a VHCDR3 of an amino acid sequence; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:501, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:502 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:503, and a VLCDR3 of the amino acid sequence of seq id no.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and TIGIT and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A TIGIT binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:496 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:497 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and TIGIT and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A TIGIT binding component comprising the amino acid sequence of SEQ ID NO:412

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and TIGIT and comprises SEQ ID NO:439 and SEQ ID NO:420.

In some embodiments, the second target epitope is an immune checkpoint inhibitor molecule. For example, in some embodiments, the second target epitope is PD-1 (programmed cell death protein 1). PD-1 is an immunomodulator antigen and is an example of a member of the immunoglobulin superfamily of cell surface receptors. PD-1 is an example of an immune checkpoint inhibitor expressed on activated CD4+/CD8+ T cells and other types of immune cells (e.g., gamma delta T cells, B cells and macrophages). Binding of PD-1 to its ligand results in inhibition of T cell activation. Under normal conditions, this plays a role in immune homeostasis: autoimmune is prevented by reducing apoptosis of tregs and increasing apoptosis of antigen-specific T cells. In the cancer setting, this can lead to immune escape of tumor cells by inactivating cytolytic cd8+ T cells. Thus, blocking PD-1 is a promising therapeutic target. Monoclonal antibodies that specifically bind to epitopes of PD-1 are well known in the art. For example, palbociclib is a monoclonal antibody that specifically binds to an epitope of PD-1.

In some embodiments, the bispecific or multispecific antibody comprises a PD-1 binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:474, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:475 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:476, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:477, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:478 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:479, and VLCDR3 of the amino acid sequence of seq id no. In some embodiments, the bispecific or multispecific antibody comprises a PD-1 binding component comprising a polypeptide comprising SEQ ID NO:472 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:473 or a VL consisting of said amino acid sequence. In some embodiments, the bispecific or multispecific antibody specifically binds to PD-1 and comprises SEQ ID NO:406.

Such sequences may be combined with any suitable anti-TRDV 1 binding sequence. For example, in some embodiments, a bispecific or multispecific antibody specifically binds to TRDV and PD-1 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no; and

-A PD-1 binding component, the PD-1 binding component comprising: a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:474, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:475 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:476, a VHCDR3 of the amino acid sequence of seq id no; and a light chain variable region comprising a light chain sequence comprising SEQ ID NO:477, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:478 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:479, and VLCDR3 of the amino acid sequence of seq id no.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV a 1 and PD-1 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A PD-1 binding component, said PD-1 binding component comprising a polypeptide comprising SEQ ID NO:472 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:473 or a VL consisting of said amino acid sequence.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV a 1 and PD-1 and comprises:

-TRDV a1 binding component, said TRDV binding component comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence; and

-A PD-1 binding component, said PD-1 binding component comprising the amino acid sequence of SEQ ID NO:406.

In some embodiments, the bispecific or multispecific antibody specifically binds to TRDV and PD-1 and comprises SEQ ID NO:438 and SEQ ID NO:417.

In some embodiments, the second target epitope is a stimulatory immune checkpoint molecule. Stimulatory immune checkpoint molecules include, for example, OX40L, 4-1BB (CD 137), CD137L, CD, CD70, CD28, GITR, ICOS, CD40 and CD40L. In some embodiments, the second target epitope is one or more selected from OX40, OX40L, 4-1BB (CD 137), CD137L, CD, CD70, CD28, GITR, ICOS, CD40, and CD40L. In some embodiments, the second target epitope is one or more selected from OX40 and 4-1 BB.

In some embodiments, the second target epitope is an immune checkpoint inhibitor molecule. Immune checkpoint inhibitor molecules include, for example, TIGIT, CD155, PD-1, PD-L1, CTLA-4, B7-H3, B7-H4, BTLA, LAG-3, VISTA and TIM-3. In some embodiments, the second target epitope is one or more selected from TIGIT, CD155, PD-1, PD-L1, CTLA-4, B7-H3, B7-H4, BTLA, LAG-3, VISTA and TIM-3. In some embodiments, the second target epitope is one or more selected from TIGIT and PD-1.

References herein to an antigen "on" a cell refer to an antigen expressed on or associated with (outside of) the cell surface membrane of such a cell.

In various embodiments, the second target epitope is an epitope of cluster of differentiation CD antigen. Cluster of Differentiation (CD) nomenclature is a unified system by which cell surface molecules can be identified and named. Typically, a cell surface protein is not assigned a CD number until at least two monoclonal antibodies directed against the cell surface protein are produced. Thus, the system ensures that all cell surface proteins assigned a CD number are readily recognized and bound by a particular monoclonal antibody or fragment thereof. In various embodiments, the CD antigen is one antigen ：CD1a、CD1b、CD1c、CD1d、CD1e、CD2、CD3、CD3d、CD3e、CD3g、CD4、CD5、CD6、CD7、CD8、CD8a、CD8b、CD9、CD10、CD11a、CD11b、CD11c、CD11d、CD13、CD14、CD15、CD16、CD16a、CD16b、CD17、CD18、CD19、CD20、CD21、CD22、CD23、CD24、CD25、CD26、CD27、CD28、CD29、CD30、CD31、CD32A、CD32B、CD33、CD34、CD35、CD36、CD37、CD38、CD39、CD40、CD41、CD42、CD42a、CD42b、CD42c、CD42d、CD43、CD44、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD50、CD51、CD52、CD53、CD54、CD55、CD56、CD57、CD58、CD59、CD60a、CD60b、CD60c、CD61、CD62E、CD62L、CD62P、CD63、CD64a、CD65、CD65s、CD66a、CD66b、CD66c、CD66d、CD66e、CD66f、CD68、CD69、CD70、CD71、CD72、CD73、CD74、CD75、CD75s、CD77、CD79A、CD79B、CD80、CD81、CD82、CD83、CD84、CD85A、CD85B、CD85C、CD85D、CD85F、CD85G、CD85H、CD85I、CD85J、CD85K、CD85M、CD86、CD87、CD88、CD89、CD90、CD91、CD92、CD93、CD94、CD95、CD96、CD97、CD98、CD99、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107、CD107a、CD107b、CD108、CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120、CD120a、CD120b、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140A、CD140B、CD141、CD142、CD143、CD144、CDw145、CD146、CD147、CD148、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD157、CD158、CD158A、CD158B1、CD158B2、CD158C、CD158D、CD158E1、CD158E2、CD158F1、CD158F2、CD158G、CD158H、CD158I、CD158J、CD158K、CD159a、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CDw198、CDw199、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218a、CD218b、CD219、CD220、CD221、CD222、CD223、CD224、CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240CE、CD240D、CD241、CD242、CD243、CD244、CD245、CD246、CD247、CD248、CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261、CD262、CD263、CD264、CD265、CD266、CD267、CD268、CD269、CD270、CD271、CD272、CD273、CD274、CD275、CD276、CD277、CD278、CD279、CD280、CD281、CD282、CD283、CD284、CD285、CD286、CD287、CD288、CD289、CD290、CD291、CD292、CDw293、CD294、CD295、CD296、CD297、CD298、CD299、CD300A、CD300C、CD301、CD302、CD303、CD304、CD305、CD306、CD307、CD307a、CD307b、CD307c、CD307d、CD307e、CD308、CD309、CD310、CD311、CD312、CD313、CD314、CD315、CD316、CD317、CD318、CD319、CD320、CD321、CD322、CD323、CD324、CD325、CD326、CD327、CD328、CD329、CD330、CD331、CD332、CD333、CD334、CD335、CD336、CD337、CD338、CD339、CD340、CD344、CD349、CD351、CD352、CD353、CD354、CD355、CD357、CD358、CD360、CD361、CD362、CD363、CD364、CD365、CD366、CD367、CD368、CD369、CD370 and CD371 selected from the group consisting of.

Healthy cell retention

While the mechanism by which γδ T cells recognize antigens and differentiate between healthy and diseased cells is not fully understood (Ming Heng and Madalene Heng,Antigen Recognition byγδT-Cells.Madame Curie Bioscience Database[Internet],Austin(TX)：Landes Bioscience;2000-2013),, γδ T cells are able to differentiate between healthy and diseased cells and exhibit significant diseased cell multicytotoxicity (see non-limiting example cell types in table 6) means that they can be used to provide improved drugs with improved therapeutic window.

TABLE 6 exemplary cancer cells killed by multicellular human V.delta.1+ cells

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As a non-limiting example, recent studies on CD3xHER2 multispecific antibodies have highlighted the challenges of current or conventional approaches. In particular, the use of such conventional methods can result in less favorable toxicity profiles. This is because, like many other tumor-associated antigens (TAAs), HER2 antigen is expressed not only in cancer (e.g., breast cancer), but also in healthy tissue (e.g., heart cells). Thus, the use of CD3xHER2 drugs that bind and co-localize all T cells to HER2 positive cells may result in a less favorable therapeutic window or therapeutic index. This is because such drugs will bind all T cells, most of which in the circulation will be αβ T cells (cd4+ positive, cd8+ positive, etc.). And once co-localized with HER2 positive cells, such conventional αβ T cells exhibit limited ability to retain her2+ healthy cells and limited ability to kill only her2+ diseased cells. Thus, by way of this example, in cynomolgus monkey studies administered such CD3xHER2 bispecific antibodies, early euthanasia (even on the day of administration) is required in some cases. Furthermore, in this example study (see Staflin et al (2020) JCI Insight 5 (7): e 133757) the conclusion was that: retargeting T cells to kill HER2 expressing cells can induce adverse effects on HER2 expressing tissues. Notably, all affected or damaged tissues, except the liver, express HER2.

In another non-limiting example, the second binding specificity may be directed against a tumor-associated moiety that also participates in controlling or modulating immune cell function. For example, the second specificity may be designed to target a so-called "checkpoint inhibitor", such as PD-L1 (CD 274) or CD155. Again, neither PDL-1 nor CD155 is 100% disease specific. Both proteins can also be expressed on healthy cells. However, multispecific antibodies designed to co-localize vδ1+ cell specificity to PD-L1 positive cells or CD155 positive cells can selectively kill PD-L1 or CD155 positive lesions or cancer cells. Further targeting disease-associated checkpoint inhibitors present on diseased cells (e.g., cancer cells) not only co-localizes vδ1+ cells to such tumors, but may also bring additional beneficial effects, such as by modulating or inhibiting PD-1/PD-L1 or TIGIT/CD155 signaling that might otherwise down-regulate T cell mediated immune responses to the disease.

Thus, instead of employing such conventional methods, provided herein are multispecific antibodies in which at least one first binding domain is capable of specifically binding to vδ1+ cells and at least one second binding domain is capable of specifically binding to targets present on diseased tissue and cells. Thus, the use of such multispecific antibodies in this manner allows vδ1+ cells to be co-localized to diseased cells that express the second target. Furthermore, given that such disease-associated targets are generally not 100% disease-specific, this approach of specifically targeting and co-locating vδ1+ effector cells may be preferred over conventional approaches. This is because vδ1+ effector cells may be able to recognize stress patterns in diseased or infected cells and thus be able to selectively kill diseased cells while retaining healthy cells that also express the same target.

Thus, the multispecific antibodies presented herein are capable of engaging the TCR of vδ1 cells, but complete activation does not occur unless tumor cells are also present. Complete engagement of the TCR by the presently presented antibodies results in partial downregulation, and it is believed that the presently presented antibody-bound vδ1 cells are fully activated and cytotoxic only in the presence of stressed cells, such as tumor cells. This is shown, for example, in fig. 50I, J, K and fig. 63A-F. This represents another important safety advantage of the methods presented herein, as off-target cytotoxicity is reduced and the full efficacy of the multispecific antibody to activate vδ1 cells is only released in the presence of tumor cells, meaning that healthy cells (even healthy cells expressing the second target antigen of the multispecific antibody) are preserved.

One mechanism behind γδ T cells being able to detect stress signals on tumor cells is thought to be due to the NCR (natural cytotoxic receptor) they express. NCR is capable of binding NCR ligands on tumor cells. Thus a dual activation mechanism can be employed in which γδ T cells are activated via TCR stimulation, including via NCR, which can sense tumor cells to allow complete activation and cytotoxicity.

This is in contrast to stimulation of αβ T cells via CD3, for example, where all stimulation is via TCR. Thus, such cells hardly distinguish between healthy cells or transformed cells, as they do not have mechanisms such as tumor cell perception (e.g. via NCR) that are independent of antigen presentation. Thus, if CD3 antibodies are Fc-effective, they will attract other immune cells, which may trigger a series of unpredictable and desirable events, such as cytokine storms, immune cell depletion, and even overactivation, leading to NK cell killing T cells, for example. In the present method, stimulation of γδ T cells with the currently presented multispecific antibodies does not raise such concerns, as γδ T cells are able to distinguish healthy cells from tumor cells, including via their NCR sensing mechanism, and thus selectively kill stressed cells, such as cancer cells or virally infected cells, due to the specificity of such diseased cells.

In another non-limiting example, the patient may have liver cancer, wherein no liver cancer specific antigen is known to be present in the patient. In this case, the second specificity of the multispecific antibody may be directed against an epitope present on many or all hepatocytes, such as asialoglycoprotein receptor 1. This will then co-localize γδ T cells to the liver, where γδ T cells can kill liver cancer cells while preserving healthy liver cells. This is shown, for example, in fig. 50I, J, K and fig. 63A-F. As a third non-limiting example, in patients with lung cancer, where it is known that there is no lung cancer antigen in the patient, the second specificity of the multispecific antibody may be directed against an epitope on a normal lung cell, such as, for example, SP-1. This will co-localize γδ T cells to the lung, where γδ T cells can kill lung cancer cells while retaining healthy lung cells. As a fourth non-limiting example, in patients with B-cell lymphoma, where it is known that there is no B-cell lymphoma antigen in the patient, the second specificity of the multispecific antibody may be directed against an epitope on normal B-cells, such as, for example, CD19. This will co-localize γδ T cells to B cells, where γδ T cells can kill lymphoma cells while retaining healthy B cells. Cell-specific antigens, cell-associated antigens, tissue-specific antigens, and tissue-associated antigens are well known in the art, and any such antigen may be targeted by the second specificity of the multispecific antibodies of the present invention.

The second binding specificity may target an antigen on the same cell as vδ1 or on a different cell of the same tissue type or a different tissue type. In certain embodiments, the target epitope may be located on different cells including different T cells, B cells, tumor cells, autoimmune tissue cells, or virus-infected cells. Alternatively, the target epitope may be located on the same cell.

The multispecific antibody or antigen-binding fragment thereof may be prepared in any format so long as the antibody or antigen-binding fragment thereof has multiple specificities. Examples of multispecific antibody formats include, but are not limited to, crossMab, DAF (two-in-one), DAF (four-in-one), dutatMab, DT-IgG, pestle-mortar structure (KIH), pestle-mortar structure (universal light chain), charge pair, fab arm exchange, SEEDbody, triomab, LUZ-Y, fcab, kappa lambda body, orthogonal Fab、DVD-IgG、IgG(H)-scFv、scFv-(H)IgG、IgG(L)-scFv、scFv-(L)IgG、IgG(L,H)-Fv、IgG(H)-V、V(H)-IgG、IgG(L)-V、V(L)-IgG、KIH IgG-scFab、2scFv-IgG、IgG-2scFv、scFv4-Ig、Zybody、DVI-IgG( four-in-one), nanobody, nanoby-HAS, biTE, diabody, DART, tandAb, scDiabody, scDiabody-CH3, diabody-CH3, triabody, morisen format, minibody, triBi minibody, scFv-CH3 KIH, fab-scFv, scFv-CH-CL-scFv, F (ab') 2, F (ab) 2-scFv2, scFv-KIH, fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intracellular antibodies, doand Lock, immTAC, HSAbody, scDiabody-ck, tandem scFv-toxin, igG-Body, duobody, mab ², ovX-scFv and 1-PEG-scFv2 (see SpiescFv et al Molecular Immunology). 95-106).

Antibodies or antigen binding fragments thereof as described herein can also be assessed by measuring their ability to enhance function in a multispecific format, e.g., a bispecific or trispecific format. Surprisingly, through such studies, it is possible to identify further functional improvements in the performance of antibodies or antigen binding fragments thereof as described herein.

The multispecific forms of various antibody sources have been previously described and are typically empirically constructed from component binding moieties. Typically, once constructed, the performance of such multi-specific or multi-target binding formats as described herein can be measured in one or more of the foregoing model systems (cell killing, cell proliferation, healthy cell retention/diseased cell specific models, etc.). Optionally, they are also compared with the component parts and other comparison molecules.

While not limited by this approach, typically when an antibody is constructed as a multispecific antibody, the binding domain module of each target (first, second, third, etc.) is optionally constructed from scFv, fab, fab ', F (ab') 2, fv, variable domains (e.g., VH or VL), diabodies, minibodies, or full-length antibodies. For example, each of the binding domains or modules is produced in one or more of the following non-limiting formats in which the binding domains comprising the variable domains and/or full length antibodies and/or antibody fragments are operably linked in tandem to produce a multispecific antibody.

Notably, multispecific antibodies comprising at least one binding domain that (first) targets the vδ1 chain of a γδ TCR as described herein are further enhanced when the first binding domain is formatted with a multispecific antibody format comprising at least one second binding domain for tissue ("solid") and hematopoietic ("liquid") disease or cell type-related targets.

Multispecific antibodies-non-limiting examples:

to outline the applicability of the method, a series of non-limiting exemplary multispecific antibodies were constructed. These multispecific antibodies comprise at least one binding domain that (first) targets the vδ1 chain of the γδ TCR and at least one binding domain that (second) targets a disease-associated target:

a first instance; vδ1-EGFR multispecific antibody:

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. The resulting dual specificity format is sometimes referred to as the 'Morisen format'. In this case, the first binding domain targets the vδ1 chain of γδ TCR and the second binding domain targets EGFR (see example 20).

A second instance; vδ1-EGFR multispecific antibody:

For this example, one binding domain (for the first target) comprises an antibody variable domain (specifically comprising VH and cognate VL domains) and the second binding domain (for the second target) comprises a binding domain within the heavy chain constant domain (CH 1-CH2-CH 3) (see also EP2546268 A1 table 1/EP3487885 A1). The resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting the EGF receptor (see example 20).

A third example; vδ1-CD19 multispecific antibody:

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting CD19 (see example 21).

A fourth example; another V.delta.1-CD 19 multispecific antibody

For this example, another multispecific antibody specificity that binds to TRDV and CD19 was prepared and tested for antigen binding (including human and cynomolgus monkey-TRDV 1), V.delta.1-cell activation, and V.delta.1-cell cytotoxicity. This example was prepared based on ADT1-4-2, see examples 26 and 27.

A fifth example; v delta 1-Her2 multispecific antibodies

For this example, multispecific antibodies that specifically bind to TRDV and Her2 were prepared and tested for antigen binding, vδ1-cell activation, and vδ1-cell cytotoxicity. This example was prepared based on ADT1-4-2, see example 28.

A sixth example; another V delta 1-EGFR multispecific antibody

For this example, another multispecific antibody that specifically binds to TRDV and EGFR was prepared and tested for antigen binding, V.delta.1-cell activation, and V.delta.1-cell cytotoxicity. This example was prepared based on ADT1-4-2, see examples 29 and 30.

A seventh example; v delta 1-FAP alpha multispecific antibodies

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting fapα (see example 33).

Eighth example; v delta 1-m Pi Suduo specific antibodies

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting mesothelin (see example 34).

A ninth example; v delta 1-PD-1 multispecific antibodies

For this example, one binding domain (for the second target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the first target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting PD-1 and a second binding domain targeting vδ1 chain of γδ TCR (see example 35).

Tenth example; v &1-4-1BB multispecific antibodies

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting 4-1BB (see example 36).

An eleventh example; v &1-OX40 multispecific antibodies

For this example, one binding domain (for the first target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the second target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting the vδ1 chain of γδ TCR and a second binding domain targeting OX40 (see example 34).

A twelfth example; v &1-TIGIT multispecific antibodies

For this example, one binding domain (for the second target) comprises the complete antibody moiety; specifically, VH-CH1-CH2-CH3 and a cognate VL-CL partner, while the second binding domain (to the first target) comprises an antibody fragment; specifically, scFv formats. The two binding modules are then fused by means of a linker. For this example, the resulting bispecific antibody comprises a first binding domain targeting TIGIT and a second binding domain targeting the vδ1 chain of γδ TCR (see example 38).

Notably, in all of the examples comprising at least one binding domain of the vδ1 chain of the (first) targeted γδ TCR and at least one second domain of the targeted second epitope, enhanced function was observed compared to the control and components (see examples 20, 21 and 26 to 38 herein).

In general, these non-limiting examples highlight the flexibility of the multispecific antibodies or antigen-binding fragments thereof as described herein. These non-limiting examples outline a multispecific antibody approach in which an antibody or fragment thereof targeting the germline V.delta.1 chain (amino acids 1-90 of SEQ ID NO: 272) can be further enhanced by combining with a second binding domain to form a multispecific antibody. As non-limiting examples, provided herein are multispecific antibodies that have enhanced functionality and contain binding domains (VH-CH 1-CH2-CH3 and VL-CL), and/or variable domains (VH and cognate VL or VH-CH1 and cognate VL-CL), and/or antibody fragments (scFv) that constitute an intact antibody.

In one embodiment, the multispecific antibody binding domain that targets the vδ1 chain (first target) of the γδ TCR may comprise (i) one or two or more antibody binding domains, each comprising a heavy chain (VH-CH 1-CH2-CH 3) and a cognate light chain partner (VL-CL), and/or (ii) one or two or more antibody binding domains, each comprising a heavy chain variable domain (VH or VH-CH 1) and a cognate light chain variable domain partner (VL or VL-VC), and/or (iii) one or two or more antibody binding domains, each comprising an antibody fragment comprising CDRs.

In one embodiment, a multispecific antibody is provided that comprises at least one first antibody-derived binding domain that targets the vδ1 chain of a γδ TCR and is operably linked to at least one second antibody binding domain that targets a second epitope. Optionally, the binding domain comprises at least one or more VH and cognate VL binding domains, or one or more VH-CH1-CH2-CH3 and cognate VL-CL binding domains, or one or more antibody fragment binding domains. Optionally, the second binding domain targets a second epitope associated with or expressed on the cell surface of the cell. Optionally, the second epitope is located on a cell surface polypeptide associated with a diseased or tumor cell or a virus-infected or autoimmune tissue cell. Optionally, the one or more second epitopes are located on disease and cell type related CD19, EGFR, her2, fapα, mesothelin, PD-1, 4-1BB, OX40 or TIGIT antigens. Optionally, the multispecific antibody comprising at least one binding domain of a first antibody source that targets the vδ1 chain of γδ TCR is operably linked to a second binding domain that binds the EGF receptor and comprises one or more of the following heavy chain modifications according to EU numbering: L358T and/or T359D and/or K360D and/or N361G and/or Q362P and/or N384T and/or G385Y and/or Q386G and/or D413S and/or K414Y and/or S415W and/or Q418Y and/or Q419K.

Optionally, the multispecific antibody comprising at least one binding domain of a first antibody source that targets the vδ1 chain of the γδ TCR is operably linked to a second binding domain comprising the amino acid sequence of SEQ ID NO:385 or SEQ ID NO:386 or SEQ ID NO:387 or SEQ ID NO:314 or SEQ ID NO:394 or SEQ ID NO:395 or SEQ ID NO:397 or SEQ ID NO:402 or SEQ ID NO:404 or SEQ ID NO:406 or SEQ ID NO:408 or SEQ ID NO:410 or SEQ ID NO:412 or a functionally equivalent binding variant thereof and targets EGFR, CD19 Her2, fapα, mesothelin, PD-1, 4-1BB, OX40 or TIGIT. Optionally, the multispecific antibody comprises SEQ ID NO:388 (388); or SEQ ID; or SEQ ID NO:315; or SEQ ID NO:316, a step of; or SEQ ID NO:378; or SEQ ID NO:379; or SEQ ID NO:380, a step of; or SEQ ID NO:382 (382); or SEQ ID NO:383; or SEQ ID NO:384; or SEQ ID NO:393; or SEQ ID NO:396; or SEQ ID NO:398; or SEQ ID NO:399 (V); or SEQ ID NO:401; or SEQ ID NO:403; or SEQ ID NO:405; or SEQ ID NO:407, a step of selecting a specific code; or SEQ ID NO:409; or SEQ ID NO:411; or SEQ ID NO:414 and SEQ ID NO:415, a base; or SEQ ID NO:414 and SEQ ID NO:416; or SEQ ID NO:414 and SEQ ID NO:419; or SEQ ID NO:414 and SEQ ID NO:418; or SEQ ID NO:504 and SEQ ID NO:415, a base; or SEQ ID NO:504 and SEQ ID NO:416; or SEQ ID NO:504 and SEQ ID NO:419; or SEQ ID NO:504 and SEQ ID NO:418; or SEQ ID NO:505 and SEQ ID NO:415, a base; or SEQ ID NO:505 and SEQ ID NO:416; or SEQ ID NO:505 and SEQ ID NO:419; or SEQ ID NO:505 and SEQ ID NO:418; or SEQ ID NO:421 and SEQ ID NO:422, a part of the material; or SEQ ID NO:504 and SEQ ID NO:422, a part of the material; or SEQ ID NO:423 and SEQ ID NO: 424. Or SEQ ID NO:425 and SEQ ID NO:426; or SEQ ID NO:421 and SEQ ID NO:437; or SEQ ID NO:504 and SEQ ID NO:437; or SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; or SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. Or SEQ ID NO:421 and SEQ ID NO:429; or SEQ ID NO:504 and SEQ ID NO:429; or SEQ ID NO:423 and SEQ ID NO:430; or SEQ ID NO:414 and SEQ ID NO:431; or SEQ ID NO:414 and SEQ ID NO:433, respectively; or SEQ ID NO:414 and SEQ ID NO:434, a base; or SEQ ID NO:414 and SEQ ID NO:435; or SEQ ID NO:504 and SEQ ID NO:431; or SEQ ID NO:504 and SEQ ID NO:433, respectively; or SEQ ID NO:504 and SEQ ID NO:434, a base; or SEQ ID NO:504 and SEQ ID NO:435; or SEQ ID NO:505 and SEQ ID NO:431; or SEQ ID NO:505 and SEQ ID NO:433, respectively; or SEQ ID NO:505 and SEQ ID NO:434, a base; or SEQ ID NO:505 and SEQ ID NO:435; or SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

Table a: summary of bispecific sequences

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In one aspect of the invention, the multispecific antibodies of the invention may be used in a therapeutically effective amount to treat a disease or disorder, thereby ameliorating at least one sign or symptom of the disease or disorder.

In one embodiment, a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of γδ TCR in a multispecific antibody format as described herein is provided, wherein the multispecific antibody is applied to vδ1+ cells to measure the effect of the multispecific entity to the vδ1+ cells (e.g. specific for the vδ1+ phenotype and/or cytotoxicity and/or diseased cells and/or enhancement thereof).

Combination therapy

The anti-vδ1 antibodies or antigen binding fragments thereof of the present invention may be used in certain combination therapies. In some embodiments, the anti-vδ1 antibodies or antigen binding fragments thereof may be combined with modulators of cancer antigens or cancer-associated antigens, for example, modulators of cancer antigens or cancer-associated antigens listed above as possible candidates for multispecific antibodies incorporating anti-vδ1 antibodies or antigen binding fragments thereof of the present invention. In some embodiments, the anti-vδ1 antibody or antigen binding fragment thereof may be combined with modulators of cluster CD antigen, for example modulators of cluster CD antigen listed above as possible candidates for multispecific antibodies incorporating the anti-vδ1 antibody or antigen binding fragment thereof of the present invention. In some embodiments, the modulator may be antagonistic or agonistic. Suitable modulators include antibodies, fusion proteins, or small molecules.

Immunoconjugates

The antibodies or antigen binding fragments thereof of the invention may be conjugated to a therapeutic moiety, such as a cytotoxin or a chemotherapeutic agent. Such conjugates may be referred to as immunoconjugates. As used herein, the term "immunoconjugate" refers to an antibody that is chemically or biologically linked to another moiety, such as a cytotoxin, a radioactive agent, a cytokine, an interferon, a target or reporter moiety, an enzyme, a toxin, a peptide or protein, or a therapeutic agent. An antibody may be linked to a cytotoxin, a radioactive agent, a cytokine, an interferon, a target or reporter moiety, an enzyme, a toxin, a peptide or a therapeutic agent at any location along the molecule as long as it is capable of binding to its target. Examples of immunoconjugates include antibody drug conjugates and antibody-toxin fusion proteins. In one embodiment, the agent may be a second, different antibody directed against vδ1. In certain embodiments, the antibody may be conjugated to an agent specific for tumor cells or virus-infected cells. The type of therapeutic moiety that can be conjugated to the anti-vδ1 antibody will take into account the condition to be treated and the desired therapeutic effect to be achieved. In one embodiment, the agent may be a secondary antibody or antigen-binding fragment thereof that binds to a molecule other than vδ1.

Affinity maturation methods and related antibodies

The invention also provides affinity matured anti-TCR delta variable 1 (anti-vδ1) antibodies or antigen-binding fragments thereof.

For example, an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof is provided, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises a polypeptide comprising SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, a VL sequence of the amino acid sequence of seq id no. In another embodiment, an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof is provided, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, a VL sequence of the amino acid sequence of seq id no. Also provided are anti-TCR delta variable 1 (anti-vδ1) antibodies or antigen-binding fragments thereof, wherein the anti-vδ1 antibodies or antigen-binding fragments thereof are affinity matured variants of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises:

Comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

Comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

Comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

Comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

Comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, and a VL of the amino acid sequence of 313.

Any known method may be used to increase the affinity of the antibodies of the invention to produce affinity matured antibodies or humanized affinity matured antibodies having increased affinity for the vδ1 antibody.

Suitably, the affinity matured variant binds to a variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) with an affinity that is at least 20%, at least 30%, at least 40%, more preferably at least 50% higher than the parent antibody, e.g. as measured by Kd.

In some embodiments, the invention provides a method of making an antibody (or antigen binding fragment thereof) of the invention, comprising providing a parent antibody disclosed herein, and subjecting the antibody to affinity maturation, wherein the produced antibody binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) with higher affinity than the parent antibody. Suitably, the produced antibody binds to the variable δ1 (vδ1) chain of the γδ T Cell Receptor (TCR) with an affinity that is at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 500% or more preferably at least about 1000% higher than the affinity of the parent antibody to the variable δ1 (vδ1) chain of the γδ T Cell Receptor (TCR), e.g. as measured by Kd. Methods of measuring affinity are known in the art and are described in the examples below. Affinity matured antibodies produced by such methods can be formulated and used as described herein for other anti-vδ1 antibodies and antigen binding fragments of the present invention.

Affinity maturation may be performed according to any suitable method known to the skilled person. For example, in vitro antibody display systems are widely used to produce specific antibodies with high affinity. In these systems, the phenotype (i.e., antibody fragment) is coupled to the genotype (i.e., antibody gene) so that the sequence of the antibody can be determined directly. Several systems have been developed to enable the display of antibody libraries to allow subsequent selection of binders and to allow selection of higher and higher affinity variants by increasing the stringency of the selection. The antibody fragments may be expressed in yeast, ribosomes, phage display particles or by direct conjugation to DNA.

Current methods of antibody affinity maturation fall into two mutagenesis categories: random and non-random. Error-prone Polymerase Chain Reaction (PCR), mutant strains and saturation mutagenesis are typical examples of random mutagenesis methods. Non-random techniques typically use alanine scanning or site-directed mutagenesis to generate a limited set of specific variants. In addition, shuffling methods that obtain shuffling variants of a parent antibody can also be used to further increase the affinity of the antibody.

Thus, in one embodiment of the invention, the method of affinity maturation is selected from the group consisting of random mutagenesis (e.g., error-prone Polymerase Chain Reaction (PCR), mutant strain or saturation mutagenesis), non-random mutagenesis (e.g., alanine scanning or site-directed mutagenesis), shuffling (e.g., DNA shuffling, strand shuffling or CDR shuffling) and introducing modifications using the CRISPR-Cas9 system.

Affinity maturation methods are described, for example, in the following: rajpal et al, proc NATL ACAD SCI USA,2005, 102 (24): 8466-71; STEINWAND et al, MAbs,2014,6 (1): 204-18; and Handbook of Therapeutic Antibodies, wiley,2014, chapter 6, antibody Affinity (pages 115-140).

In some embodiments, a method of preparing a pharmaceutical composition is provided, the method comprising providing an antibody prepared according to the above method (i.e., producing an antibody by affinity maturation), and co-formulating the antibody with at least one or more pharmaceutically acceptable excipients. Antibodies used in the preparation of the pharmaceutical compositions may be affinity matured variants of G04, E07, C08, B07, C05, E04, F07, G06, G09, B09, G10 or E01, including any and all ADT1-4 and ADT1-7 lineage antibodies and antigen binding fragments and variants thereof. Pharmaceutical compositions produced by such methods can be used in the methods of treatment of the invention as described herein for other anti-vδ1 antibodies.

Thus, provided are anti-vδ1 antibodies and antigen binding fragments thereof, which are affinity matured mutants or variants of the antibodies disclosed herein. For example, in one embodiment, affinity matured variants of an antibody selected from the group consisting of: g04, E07, C08, B07, C05, E04, F07, G06, G09, B09, G10, and E01. In general, affinity maturation mutants have higher affinity for the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) than the parent antibody (the antibody from which the mutant is derived). The invention also provides antibodies and antigen-binding fragments thereof obtainable or obtained by affinity maturation of an antibody or antigen-binding fragment thereof of the invention.

The method of affinity maturation may preferentially introduce favorable mutations into specific parts of the parent sequence. For example, in some embodiments, the affinity matured variant retains certain regions of the parent antibody (or the methods of the invention do not introduce mutations into certain regions of the parent antibody). For example, in some embodiments, the affinity maturation variant retains:

a) HCDR2, LCDR1 and LCDR2 sequences (e.g., without limitation, antibodies derived from G04)

B) HCDR1, HCDR2, LCDR1 and LCDR2 sequences (e.g., but not limited to antibodies derived from E07)

C) HFR2, HFR3, HFR4, LFR1, and LFR2 sequences (e.g., without limitation, antibodies derived from G04)

D) HFR1, HFR2, HFR3, HFR4, LFR2, LFR3, and LFR4 sequences (e.g., without limitation, antibodies derived from E07)

E) HFR2, HCDR2, HFR3, HFR4, LFR1, LCDR1, LFR2, and LCDR2 sequences (such as, but not limited to, antibodies derived from G04); or (b)

F) HFR1, HCDR1, HFR2, HCDR2, HFR3, HFR4, LCDR1, LFR2, LCDR2, LFR3, and LFR4 sequences (such as, but not limited to, antibodies derived from E07).

In some embodiments, the affinity matured antibody comprises a kappa light chain variable sequence but comprises a non-serine residue at residue position 74 according to the IMGT numbering system of the light chain variable sequence. Typically, the amino acid at this residue is a naturally occurring amino acid. In some embodiments, the residue at the position is a non-polar residue (e.g., any amino acid selected from the group consisting of glycine, alanine, valine, methionine, leucine, and isoleucine) and/or a non-human germline residue at the position (e.g., any amino acid selected from the group consisting of arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine). In some embodiments, the amino acid residue at position 74 is leucine.

In some embodiments, the affinity matured antibody retains the HFR2, HCDR2, HFR3, HFR4, LFR1, LCDR1, LFR2, LCDR2, and HFR3 sequences of the parent antibody (e.g., but not limited to an antibody derived from G04), amino acids other than serine (e.g., non-polar and/or non-human germline residues at position 74 in the LFR3 region, such as leucine residues).

In some embodiments, affinity matured antibodies retain the HFR1, HCDR1, HFR2, HCDR2, HFR3, HFR4, LCDR1, LFR2, LCDR2, LFR3, and LFR4 sequences of the parent antibody (e.g., but not limited to, an antibody derived from E07), except that position 74 in the LFR3 region is mutated to a non-serine residue (e.g., a non-human germline and/or a non-polar residue at position 74, such as a leucine residue).

In some embodiments, the affinity matured antibody retains a certain percentage of identity compared to the parent sequence. For example, in some embodiments, the affinity matured antibody comprises VH and VL sequences that are at least 80%, at least 90%, at least 95%, or at least 96% identical to the corresponding parent VH and VL sequences.

In some embodiments, the affinity maturation variant retains:

a) HCDR2, LCDR1 and LCDR2 sequences (such as but not limited to antibodies derived from G04) and comprise VH and VL sequences at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences;

b) HCDR1, HCDR2, LCDR1 and LCDR2 sequences (such as but not limited to antibodies derived from E07) and comprise VH and VL sequences at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences;

c) HFR2, HFR3, HFR4, LFR1 and LFR2 sequences (such as but not limited to antibodies derived from G04) and comprise VH and VL sequences that are at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences;

d) HFR1, HFR2, HFR3, HFR4, LFR2, LFR3, and LFR4 sequences (such as, but not limited to, antibodies derived from E07) and comprise VH and VL sequences at least 80%, at least 90%, or at least 95%, or at least 96% identical to the corresponding parent VH and VL sequences;

e) HFR2, HCDR2, HFR3, HFR4, LFR1, LCDR1, LFR2, and LCDR2 sequences (such as, but not limited to, antibodies derived from G04); and comprising VH and VL sequences at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences; or (b)

F) HFR1, HCDR1, HFR2, HCDR2, HFR3, HFR4, LCDR1, LFR2, LCDR2 LFR3 and LFR4 sequences (such as but not limited to antibodies derived from E07) and comprise VH and VL sequences at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences.

In some embodiments, affinity matured antibodies retain the HFR2, HCDR2, HFR3, HFR4, LFR1, LCDR1, LFR2, LCDR2, and HFR3 sequences of a parent antibody (e.g., but not limited to an antibody derived from G04), except that position 74 in the LFR3 region is mutated to a non-serine residue (e.g., a non-human germline and/or non-polar residue at position 74, such as a leucine residue), and the antibody is at least 80%, at least 90%, or at least 95%, or at least 96% identical VH and VL sequences to the corresponding parent VH and VL sequences.

In some embodiments, the affinity matured antibody retains the HFR1, HCDR1, HFR2, HCDR2, HFR3, HFR4, LCDR1, LFR2, LCDR2, LFR3, and LFR4 sequences of a parent antibody (e.g., but not limited to an antibody derived from E07), except that position 74 in the LFR3 region is mutated to a non-serine residue (e.g., a non-human germline and/or a non-polar residue at position 74, e.g., a leucine residue), and the antibody comprises VH and VL sequences that are at least 80%, at least 90% or at least 95% or at least 96% identical to the corresponding parent VH and VL sequences.

In some embodiments, the affinity matured antibody comprises up to 20, e.g., up to 15, e.g., up to 10 amino acid substitutions compared to the parent antibody sequence.

Specific methods can be used to prepare affinity matured antibodies. For example, affinity matured antibodies of the invention are prepared according to a particular classification scheme to help provide affinity matured antibodies with particularly advantageous properties. Specifically, all anti-Vd 1 mabs were analyzed for binding to the native target γδ TCR expressed by primary γδ T cells. Antibody binding to γδ T cells was tested by incubating a fixed concentration of purified antibody with 3×10 ζ T cells of skin origin. This incubation is performed under blocking conditions to prevent non-specific binding of the antibody via the Fc receptor. Detection was performed by adding a fluorescent dye conjugated secondary antibody to human IgG 1. For negative controls, cells were prepared as follows: a) Only isotype antibodies (recombinant human IgG), b) only fluorochrome conjugated anti-human IgG antibodies. As positive controls, parental clones and commercial anti-Vd 1 mAB clone TS8.2 were included. Most affinity matured antibodies showed improved binding to the native target compared to commercial anti-vδ1 tool antibodies, as indicated by the increased signal amplitude (MFI) and the percentage of vδ1 cells detected in the skin-derived γδt cell heterogeneous population.

To further classify anti-vδ1 mAb, 12 antibodies from each library with the best improved dissociation rate parameters compared to the parental clones were used for full KD assays by SPR with human and cynomolgus monkey antigens for ADT1-4 lineage and human antigens only for ADT1-7 lineage. 8 clones of the ADT1-4 lineage (ADT 1-4-19, ADT1-4-21, ADT1-4-31, ADT1-4-53, ADT1-4-2, ADT1-4-86, ADT1-4-112 and ADT 1-4-143) and 3 clones of the ADT1-7 clone (ADT 1-7-20, ADT1-7-3, ADT 1-7-61) were then used for functional characterization. These clones (as well as ADT1-4-1, ADT1-4-6 and ADT 1-4-138) were characterized in a TCR down-regulation assay to confirm improved target engagement compared to the parental clones. The improvement in activating vδ1 cells was tested in an up-regulation assay of CD107a in an imaging-based killing assay to test for the ability of the affinity matured antibodies to increase cytotoxicity of skin-derived vδ1γδt cells against cancer cells.

Thus, in some embodiments, a method of making an affinity matured anti-vδ1 antibody may comprise the steps of:

a) Providing a parent anti-vδ1 antibody having VH and VL region sequences;

b) Providing a set of one or more anti-vδ1 antibody variants derived from a parent antibody, wherein the VH region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VH region of the parent antibody and the VL region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VL region of the parent antibody;

c) The set of antibody variants is screened to identify antibody variants having a higher affinity for human and/or cynomolgus TRDV1 than the parent anti-vδ1 antibody to provide a subset of antibody variants having a greater affinity for human and/or cynomolgus TRDV1 than the parent antibody. Such a screening step can be performed by testing the binding of the panel of antibody variants to γδ TCRs expressed by primary γδ T cells. Additionally or alternatively, such a screening step may comprise determining KD values for one or more antibody variants in the set, for example, by surface plasmon resonance. In some embodiments, antibody variants may be selected for use in a subgroup when their affinity for human and/or cynomolgus TRVD1 is at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 500% or at least about 1000% higher than the affinity of the parent antibody.

Sequence variation between the parent antibody and one or more antibody variants in the first set may occur anywhere in the variable region sequence. In some embodiments, sequence variation between the parent antibody and one or more antibody variants in the first set may occur in CDR regions.

The method may further comprise:

d) Functionally screening a subset of antibody variants.

Functionally screening the subset of antibody variants may include screening the antibody variants for a polypeptide that causes:

TCR down-regulation in vδ1 cells;

increased NCR expression in vδ1 cells;

increased 4-1BB expression in V.delta.1 cells;

Up-regulation of CD107a in vδ1 cells; and/or

V.V delta 1 cell mediated killing of cancer cells; optionally, a plurality of

V delta 1 cell depletion.

The method may further comprise:

e) One or more antibody variants are selected from a subset of antibody variants to provide one or more affinity matured anti-vd 2 antibodies.

The selection criteria may be:

i. causing TCR down-regulation in vδ1 cells;

causing an increase in NCR expression in vδ1 cells;

causing an increase in 4-1BB expression in vδ1 cells;

Causing up-regulation of CD107a in vδ1 cells; and/or

V. causing vδ1 cell mediated killing of cancer cells; optionally, a plurality of

Does not cause the depletion of vδ1 cells.

In some embodiments, the method may be a method of providing one or more affinity matured anti-vδ1 antibodies, the method comprising:

a) Providing a parent anti-vδ1 antibody having VH and VL region sequences;

b) Providing a set of one or more anti-vδ1 antibody variants derived from a parent antibody, wherein the VH region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VH region of the parent antibody and the VL region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VL region of the parent antibody;

c) The set of antibody variants is screened to identify antibody variants having an affinity for human TRDV1 that is at least 500% higher than the affinity of the parent anti-vδ1 antibody to provide a subset of antibody variants.

D) Functionally screening a subset of antibody variants to identify antibody variants in the subset as follows:

i. causing TCR down-regulation in vδ1 cells;

causing an increase in NCR expression in vδ1 cells;

causing an increase in 4-1BB expression in vδ1 cells;

Causing up-regulation of CD107a in vδ1 cells; and/or

Inducing vδ1 cell mediated killing of cancer cells

E) Selecting one or more antibody variants from a subset of antibody variants to provide one or more affinity matured anti-vδ1 antibodies, wherein the one or more affinity matured anti-vδ1 antibodies:

i. causing TCR down-regulation in vδ1 cells;

causing an increase in NCR expression in vδ1 cells;

causing an increase in 4-1BB expression in vδ1 cells;

Causing up-regulation of CD107a in vδ1 cells; and/or

Inducing vδ1 cell mediated killing of cancer cells

In some embodiments, the method may be a method of providing one or more affinity matured anti-vδ1 antibodies, the method comprising:

a) Providing a parent anti-vδ1 antibody having VH and VL region sequences;

b) Providing a set of one or more anti-vδ1 antibody variants derived from a parent antibody, wherein the VH region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VH region of the parent antibody and the VL region of each of the one or more antibody variants comprises a sequence at least about 96% identical to the VL region of the parent antibody;

c) Screening the set of antibody variants to identify antibody variants having an affinity for human TRDV1 that is at least 500% higher than the affinity of the parent anti-vδ1 antibody;

d) Screening the set of antibody variants to identify antibody variants having an affinity for cynomolgus TRDV1 that is at least 500% higher than the affinity of the parent anti-vδ1 antibody;

e) Providing a subset of anti-vδ1 antibody variants, wherein the subset of anti-vδ1 antibody variants comprises anti-vδ1 antibody variants that bind to human and cynomolgus monkey TRDV1 with an affinity that is at least 500% higher than the affinity of the parent anti-vδ1 antibody for human and cynomolgus monkey TRDV 1;

f) Functionally screening a subset of antibody variants to identify antibody variants in the subset as follows:

i. causing TCR down-regulation in vδ1 cells;

causing an increase in NCR expression in vδ1 cells;

causing an increase in 4-1BB expression in vδ1 cells;

Causing up-regulation of CD107a in vδ1 cells; and/or

Inducing vδ1 cell mediated killing of cancer cells

G) Selecting one or more antibody variants from a subset of antibody variants to provide one or more affinity matured anti-vδ1 antibodies, wherein the one or more affinity matured anti-vδ1 antibodies:

i. causing TCR down-regulation in vδ1 cells;

causing an increase in NCR expression in vδ1 cells;

causing an increase in 4-1BB expression in vδ1 cells;

Causing up-regulation of CD107a in vδ1 cells; and/or

Causing vδ1 cell mediated killing of cancer cells.

Cynomolgus monkey cross-reactivity

Suitably, the anti-vδ1 antibody and antigen binding fragment thereof exhibit a sequence substantially identical to human TRDV SEQ ID NO:272 (including polymorphic variants, SEQ ID NO: 306) and cynomolgus monkey TRDV1 (SEQ ID NO: 308). Cross-reactivity clearly helps to provide antibodies that can be used in vivo animal studies during preclinical evaluation.

Unexpectedly, the inventors identified framework mutations that could confer increased binding of antibodies, such as anti-vδ1 antibodies and antigen binding fragments thereof, to cynomolgus monkey antigens. The framework mutations do not adversely affect the affinity of the antibody or antigen binding fragment thereof for the corresponding human version of the antigen. The mutation is based on mutation of the serine residue at position 74 of the IMGT numbering system of the kappa light chain variable sequence to a non-serine residue (e.g., non-human germline and/or non-polar residues at position 74. Non-polar amino acids may be selected from the group consisting of glycine, alanine, valine, methionine, leucine and isoleucine. Non-germline amino acids may be selected from the group consisting of arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine and valine. Non-polar and non-germline amino acids (i.e., non-polar and non-germline amino acids) may be selected from the group consisting of glycine, valine, methionine, leucine and isoleucine). In some embodiments, the mutation is a mutation of a serine residue to a leucine residue according to IMGT numbering system position 74 of the kappa light chain variable sequence. Mutations are direct substitutions, so the total length of the chain is unchanged. Thus, according to the IMGT numbering system, serine at position 74 is removed and directly replaced with another amino acid (e.g. a non-human germline and/or a non-polar amino acid at position 74, e.g. leucine). Substitution may be accomplished according to any suitable method known to the skilled artisan.

For example, fig. 26 shows this. Figure 26 demonstrates the significant effect conferred by mutating kappa chain serine 74 to leucine. In the kappa lines, position 74 is only partially conserved, and interestingly there are many different polar/non-polar/charged amino acids in different kappa chain line subclasses (see imgt. Org). However, to our knowledge, this is the first example of any kappa chain with leucine at position 74. Regardless, this study ultimately demonstrates that by mutating this single amino acid position, the affinity of the ADT1-4 parent molecule for human and cynomolgus monkey vδ1 TCR can be enhanced by about 13-fold and 5-fold, respectively. To further verify this finding, leucine 74 was reverted to the starting germline serine for the high affinity exemplified sub-molecule (ADT 1-4-2). In contrast, this demonstrates that by reverting back to germline serine, the affinity for human and cynomolgus monkey vδ1 can be reduced by about 11-fold and 23-fold, respectively (see fig. 26D (i) and (ii)). These results dissect the importance of the additive, ratcheting role of CDR3 modification in combination with kappa light chain position 74 modification when the affinity of ADT1-4 for human and cynomolgus vδ 1 TCR was enhanced by 100-fold or more to optimal. These findings are non-exhaustive examples of how to (i) enhance the affinity of the parent ADT1-4 molecule or (ii) reduce the affinity of the high affinity sub-molecule (as shown by ADT1-4-2 in this example). Thus, these findings underscores that one can modulate the affinity of such molecules up and down as desired by making targeted combinatorial changes to heavy and light chain CDR3, in combination with selection of specific amino acids at kappa chain position 74.

References herein to antibodies comprising a kappa light chain variable sequence comprising a non-serine residue at position 74 according to IMGT numbering are alternatively defined as antibodies comprising a light chain variable sequence comprising an LFR1 region, an LCDR1 region, an LFR2 region, an LCDR2 region, an LFR3 region, an LCDR3 region and an LFR4 region, wherein the LFR3 region comprises a non-serine residue at position 74 according to IMGT numbering (e.g., a non-human germline and/or a non-polar amino acid at position 74, e.g., a leucine residue). This alternative definition applies to all antibodies disclosed herein comprising a kappa light chain variable sequence comprising a non-serine residue (e.g., a non-human germline and/or a non-polar amino acid such as a leucine residue) at position 74 according to IMGT numbering.

Accordingly, the present invention provides an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising a kappa light chain variable sequence, wherein the residue at position 74 of the light chain variable sequence is non-serine (e.g. at non-human germline and/or non-polar amino acid position 74, e.g. a leucine residue), according to the IMGT numbering system. In some embodiments, the residue at position 74 of the light chain variable sequence is leucine according to the IMGT numbering system. Suitably, the antibody may be an IgG antibody. For example, the antibody may be an IgG1 antibody.

The anti-vδ1 antibodies and antigen binding fragments thereof provided herein may possess a substitution at position 74 of the light chain variable sequence (according to the IMGT numbering system). For example, the anti-vδ1 antibodies and antigen-binding fragments and variants thereof described herein may comprise a kappa light chain variable sequence comprising residues other than serine at position 74 (e.g., non-human germline and/or non-polar amino acids such as leucine residues at position 74) according to the IMGT numbering system. In some embodiments, the anti-vδ1 antibodies and antigen-binding fragments and variants thereof described herein may comprise a kappa light chain variable sequence comprising a leucine residue at position 74 according to the IMGT numbering system. Embodiments comprising a mutation at position 74 according to the IMGT numbering system of the kappa light chain may be particularly relevant for antibodies derived from ADT1-4, although the mutation may be equally applicable to other antibodies. For example, light chain position 74 was observed to be not highly conserved among different kappa/lambda light chain germlines. Furthermore, it is noted that different light chain germlines contain different polarized and/or charged amino acids at this position, ranging from non-polar (e.g., alanine) to polar neutral (e.g., serine) to negatively or positively charged (e.g., aspartic acid or asparagine, respectively). It is well known that the polarity/charge of an amino acid at any given position may affect the protein structure. For example, it is well known that changes in polarity and charge affect hydrophobicity and the tendency of amino acids to be more surface exposed or more buried. Regardless, this remarkable finding underscores that non-conservative amino acid changes in light chain position 74 can alter affinity for a selected target by more than a factor of 10. Thus, in addition to increasing affinity as outlined herein, the use of this knowledge may also provide a new approach to increasing or decreasing affinity more broadly. For example, when one wishes to increase or decrease affinity for any given antibody, a method involving changing the residue at position 74 from more polar to non-polar, or for example changing non-polar to more charged, may be more preferred than a complex/cumbersome mutagenesis method such as saturation mutagenesis.

In another embodiment of the invention, there is also provided a method of mutating an antibody or antigen binding fragment thereof, the method comprising providing an antibody comprising a kappa light chain having serine at position 74 of the light chain variable sequence according to the IMGT numbering system and mutating serine to a different residue, e.g. a non-human germline and/or a non-polar amino acid, e.g. leucine. In some embodiments, the antibody is an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof. In some embodiments, the antibody is an antibody having a sequence comprising a sequence identical to SEQ ID NO:1, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO:26, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

In some embodiments, the antibody is an antibody having a sequence comprising a sequence identical to SEQ ID NO:106 at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:118 at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

In some embodiments, the antibody is an antibody comprising:

Comprising a sequence identical to SEQ ID NO:273, and VH comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:282, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:274, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:283, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:275, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:284, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:276, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:285 at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:277, and VH comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:286, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:278, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:287, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:279, and VH comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:288, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:280, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:289, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

Comprising a sequence identical to SEQ ID NO:281 at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO:290, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence. Or (b)

Comprising a sequence identical to SEQ ID NO:312, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO:313, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical amino acid sequence.

In some embodiments, the antibody is an antibody comprising:

Comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

Comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

Comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

Comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

Comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, and a VL of the amino acid sequence of 313.

Antibodies produced by this method have 100% identity to the specified VH and VL sequences, except for the mutation at position 74.

In some embodiments, the mutation at position 74 increases the affinity of the antibody or antigen binding fragment thereof for a cognate cynomolgus monkey (cynomolgus monkey) antigen. The increase in affinity of the antibody or antigen binding fragment thereof is relative to the affinity of the antibody or antigen binding fragment thereof prior to introduction of the mutation (when measured under the same or substantially the same conditions). In some embodiments, the antibody or antigen binding fragment thereof is an anti-TCR delta variable 1 (anti-V δ1) antibody or antigen binding fragment thereof that binds to a variable δ1 (V δ1) chain of a human γδ T Cell Receptor (TCR) (e.g. SEQ ID NO:272 and/or 306) after the mutation is introduced, and the antibody has increased affinity for a variable δ1 (V δ1) chain of a cynomolgus γδ T Cell Receptor (TCR) (e.g. SEQ ID NO: 308) after the mutation is introduced.

In any embodiment, the amino acid at position 74 of the light chain variable sequence (according to IMGT numbering system) may preferably be leucine.

Polynucleotide sequences and expression vectors

In one aspect of the invention, polynucleotides encoding the anti-vδ1 antibodies or multispecific antibodies or fragments of the invention are provided. In one embodiment, the polynucleotide comprises a nucleotide sequence that hybridizes to SEQ ID NO:199 to 222, 224 to 247, 249 to 259 or 261 to 271, has or consists of a sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity. In another embodiment, the polynucleotide comprises SEQ ID NO:199 to 222, 224 to 247, 249 to 259 or 261 to 271 or consists of said sequence. In another aspect, a cDNA comprising the polynucleotide is provided.

In one aspect of the invention, there is provided a polynucleotide comprising a sequence that hybridizes to SEQ ID NO:199 to 222, 224 to 247, 249 to 259 or 261 to 271, has or consists of a sequence of at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity, encoding CDR1, CDR2 and/or CDR3 of the encoded immunoglobulin chain variable domain.

In one aspect of the invention, there is provided a polynucleotide comprising a sequence that hybridizes to SEQ ID NO:199 to 222, 224 to 247, 249 to 259, or 261 to 271, has or consists of a sequence of at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity, encoding FRI, FR2, FR3, and/or FR4 of the encoded immunoglobulin chain variable domain.

The invention also provides expression vectors and plasmids comprising the polynucleotide sequences of the invention. In some embodiments, the expression vector comprises a nucleotide sequence that hybridizes to SEQ ID NO:199 to 222 or 249 to 259 has at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity or a sequence (encoding a variable heavy chain region) that is 100% identical. In some embodiments, the expression vector comprises a nucleotide sequence that hybridizes to SEQ ID NO:224 to 247 or 261 to 271 has at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity or a sequence (encoding a variable light chain region) that is 100% identical. Such expression vectors may be used in pairs to pair heavy and light chain variable sequences appropriately according to the pairing of the various amino acid sequences providing the antibodies of the invention disclosed herein. In some embodiments, the expression vector comprises a nucleotide sequence that hybridizes to SEQ ID NO:199 to 222 or 249 to 259, and further comprises a sequence (encoding a variable heavy chain region) having at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity or 100% identity to SEQ ID NO:224 to 247 or 261 to 271 has at least 70%, such as at least 80%, such as at least 90%, such as at least 95%, such as at least 99% sequence identity or a sequence (encoding a variable light chain region) that is 100% identical. Likewise, sequences may be provided in specific pairs as described herein to encode antibodies of the invention.

The invention also provides polynucleotide sequences encoding all antibody sequences disclosed herein, including any variant antibody sequences disclosed herein optionally comprising one or more amino acid substitutions, as well as expression vectors and plasmids.

The polynucleotides and expression vectors of the invention may also be described with reference to the encoded amino acid sequences. Thus, in one embodiment, the polynucleotide comprises a sequence encoding SEQ ID NO:1 to 505 or consists of the sequence of the amino acid sequence of any one of claims 1 to 505.

To express an antibody or antigen-binding fragment thereof, polynucleotides encoding portions or full length light and heavy chains as described herein are inserted into an expression vector such that the genes are operably linked to transcriptional and translational control sequences. Thus, in one aspect of the invention, there is provided an expression vector comprising a polynucleotide sequence as defined herein. In one embodiment, the expression vector comprises SEQ ID NO:199 to 222 or any of 249 to 259. In another embodiment, the expression vector comprises SEQ ID NO:224 to 247 or 261 to 271.

It will be appreciated that the nucleotide sequences described herein may comprise additional sequences encoding amino acid residues to aid translation, purification and detection, however alternative sequences may be used depending on the expression system used. These optional sequences may be removed, modified or substituted if alternative design, translation, purification or detection strategies are employed.

The DNA or cDNA encoding the polypeptide may be subjected to mutations that are silent with respect to the amino acid sequence of the polypeptide, but provide preferred codons for translation in a particular host. Preferred codons for translating nucleic acids, for example in E.coli and Saccharomyces cerevisiae, as well as in mammals, in particular in humans, are known.

Mutation of a polypeptide may be achieved, for example, by substitution, addition or deletion of a nucleic acid encoding the polypeptide. Substitutions, additions or deletions to the nucleic acid encoding the polypeptide may be introduced by a number of methods including, for example, error-prone PCR, shuffling, oligonucleotide-directed mutagenesis, assembly PCR, PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive ensemble mutagenesis, exponential ensemble mutagenesis, site-specific mutagenesis, gene recombination, artificial gene synthesis, gene Site Saturation Mutagenesis (GSSM), synthetic Ligation Recombination (SLR), or a combination of these methods. Modifications, additions or deletions to the nucleic acid may also be introduced by methods including: recombination, recursive sequence recombination, phosphorothioate modified DNA mutagenesis, uracil-containing template mutagenesis, gap double mutagenesis, point mismatch repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiation-induced mutagenesis, deletion mutagenesis, restriction-selection mutagenesis, restriction-purification mutagenesis, ensemble mutagenesis, chimeric nucleic acid multimer creation, or a combination thereof.

Specifically, artificial gene synthesis may be used. Genes encoding polypeptides of the invention may be produced synthetically, for example, by solid phase DNA synthesis. The entire gene can be synthesized de novo, eliminating the need for a precursor template DNA. To obtain the desired oligonucleotides, building blocks (building blocks) are sequentially coupled to the growing oligonucleotide chains in the order required for the product sequence. After chain assembly is complete, the product is released from the solid phase into solution, deprotected and collected. The product may be separated by High Performance Liquid Chromatography (HPLC) to obtain the desired oligonucleotide in high purity.

Expression vectors include, for example, plasmids, retroviruses, cosmids, yeast Artificial Chromosomes (YACs) and epstein-barr virus (EBV) derived episomes. The polynucleotides are ligated into a vector such that transcriptional and translational control sequences in the vector perform their intended function of regulating the transcription and translation of the polynucleotide. Expression and/or control sequences may include promoters, enhancers, transcription terminators, initiation codons 5' relative to the coding sequence (i.e., ATG), splicing signals for introns, and termination codons. The expression vector and expression control sequences are selected to be compatible with the expression host cell used. For example, the sequence may comprise a nucleotide sequence encoding a single-chain variable fragment of an antibody of the invention, comprising a VH region and a VL region joined by a synthetic linker (e.g., encoding SEQ ID NO: 291). It will be appreciated that the polynucleotides or expression vectors of the invention may comprise VH regions, VL regions, or both (optionally including linkers). Thus, the polynucleotides encoding the VH and VL regions may be inserted into separate vectors, alternatively, the sequences encoding the two regions may be inserted into the same expression vector. One or more polynucleotides are inserted into an expression vector by standard methods (e.g., ligating the polynucleotide and complementary restriction sites on the vector, or making blunt-ended ligation if no restriction sites are present).

A convenient vector is one that encodes a functionally complete human CH or CL immunoglobulin sequence, with appropriate restriction sites engineered so that any VH or VL sequence can be easily inserted and expressed, as described herein. The expression vector may also encode a signal peptide that facilitates secretion of the antibody (or antigen binding fragment thereof) from the host cell. The polynucleotide may be cloned into a vector such that the signal peptide is linked in frame to the amino terminus of the antibody. The signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

In one aspect of the invention, there is provided a cell (e.g. a host cell, such as a recombinant host cell) comprising a polynucleotide or expression vector as defined herein. It will be appreciated that the cell may comprise a first vector encoding the light chain of the antibody or antigen-binding fragment thereof, and a second vector encoding the heavy chain of the antibody or antigen-binding fragment thereof. Alternatively, both the heavy and light chains are encoded on the same expression vector that is introduced into the cell.

In one embodiment, the polynucleotide or expression vector encodes a membrane-anchored or transmembrane domain fused to an antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is presented on the cell's outer surface.

Transformation may be performed by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of one or more polynucleotides in liposomes, gene gun injection, and direct microinjection of DNA into the nucleus. In addition, the nucleic acid molecules may be introduced into mammalian cells by viral vectors.

Mammalian cell lines useful as hosts for expression are well known in the art and include a number of immortalized cell lines available from the American Type Culture Collection (ATCC). These cell lines include, inter alia, chinese Hamster Ovary (CHO) cells, NSO, SP2 cells, heLa cells, baby Hamster Kidney (BHK) cells, monkey kidney Cells (COS), human hepatocellular carcinoma cells (e.g., hep G2), a549 cells, 3T3 cells, and many other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cow, horse, and hamster cells. Particularly preferred cell lines are selected by determining which cell lines have high expression levels. Other cell lines that may be used are insect cell lines (e.g. Sf9 cells), amphibian cells, bacterial cells, plant cells and fungal cells. Antigen binding fragments of antibodies, such as scFv and Fv fragments, can be isolated and expressed in e.coli using methods known in the art.

Antibodies are produced by culturing the host cells for a period of time sufficient to allow expression of the antibodies in the host cells, or more preferably secretion of the antibodies into the medium in which the host cells are grown. Antibodies can be recovered from the culture medium using standard protein purification methods.

Antibodies (or fragments) of the invention may be used, for example, with Green and Sambrook, molecular Cloning: a Laboratory Manual (2012) 4 th edition Cold Spring Harbour Laboratory Press.

Monoclonal antibodies can be produced using hybridoma technology by fusing specific B cells that produce the antibody with myeloma (B cell carcinoma) cells, which are selected for their ability to grow in tissue culture and the absence of antibody chain synthesis.

Monoclonal antibodies directed against the defined antigens can be obtained, for example, by:

a) Immortalized lymphocytes obtained from the peripheral blood of an animal previously immunized with the defined antigen, with immortalized cells and preferably with myeloma cells, so as to form hybridomas,

B) The immortalized cells (hybridomas) formed are cultured and the cells producing antibodies of the desired specificity are recovered.

Or the use of hybridoma cells is not required. Antibodies capable of binding to a target antigen as described herein may be isolated from a suitable antibody library via conventional practice, e.g., using phage display, yeast display, ribosome display, or mammalian display techniques known in the art. Thus, monoclonal antibodies can be obtained, for example, by a process comprising the steps of:

a) Cloning DNA or cDNA sequences obtained from lymphocytes, in particular peripheral blood lymphocytes of animals (suitably previously immunized with defined antigens), into vectors, in particular phages and more particularly filamentous phages,

B) Prokaryotic cells are transformed with the above vectors under conditions that allow antibody production,

C) Antibodies are selected by subjecting the antibodies to antigen affinity selection,

D) Recovering the antibody with the desired specificity.

Optionally, an isolated polynucleotide encoding an antibody or antigen binding fragment thereof as described herein and binding to the vδ1 chain of γδ can also be readily manufactured in an amount sufficient to produce a medicament for use as an improvement in the sign or symptom of a disease. When used as a medicament in this manner, the polynucleotide of interest is typically first operatively linked to an expression vector or cassette designed to express the antibody or antigen-binding fragment thereof in a subject or patient. Such expression cassettes and methods of delivering polynucleotides or sometimes referred to as 'core-based' drugs are well known in the art. For a recent review, see Hollevoet and Declerck (2017) j. Transl. Med.15 (1): 131.

Also provided is a method for producing an anti-vδ1 antibody or antigen-binding fragment or variant thereof comprising culturing a host cell of the present invention in a cell culture medium under conditions for intracellular expression of a nucleic acid sequence encoding a plasmid or vector. The method may further comprise obtaining an anti-vδ1 antibody or antigen binding fragment or variant thereof from the cell culture supernatant. The resulting antibodies can then be formulated into pharmaceutical compositions. Further, there is provided a method of producing a cell expressing an anti-vδ1 antibody or antigen binding fragment or variant thereof, the method comprising transfecting the cell with a plasmid or vector of the present invention. The cells may then be cultured to produce an anti-vδ1 antibody or antigen binding fragment or variant thereof.

Pharmaceutical composition

According to a further aspect of the present invention there is provided a composition comprising an antibody or antigen binding fragment thereof as defined herein. In such embodiments, the composition may comprise antibodies optionally in combination with other excipients. Also included are compositions comprising one or more additional active agents (e.g., active agents suitable for treating the diseases mentioned herein).

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising an antibody or antigen binding fragment thereof as defined herein together with a pharmaceutically acceptable diluent or carrier. The antibodies of the invention may be incorporated into a pharmaceutical composition suitable for administration to a subject. Typically, the pharmaceutical composition comprises an antibody of the invention and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. In many cases, it will be preferable to include an isotonic agent, for example, sugars, polyalcohols (e.g., mannitol, sorbitol) or sodium chloride in the composition. Pharmaceutically acceptable substances, such as wetting agents or minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives or buffers, may be included which enhance the shelf life or effectiveness of the antibody or antigen binding fragment thereof.

The compositions of the present invention may take a variety of forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The preferred form depends on the intended mode of administration and the therapeutic application. Typically preferred compositions are in the form of injectable or infusible solutions.

The preferred mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular, intrathecal). In a preferred embodiment, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

Therapeutic compositions must generally be sterile and stable under the conditions of manufacture and storage. The compositions may be formulated as solutions, microemulsions, dispersions, liposomes or other ordered structures suitable for high drug concentrations.

The pharmaceutical compositions of the invention are within the scope of the invention as an adjunct to or in combination with other established therapies typically used to treat diseases as described herein in therapeutic methods for treating such diseases.

In another aspect of the invention, the antibody, composition or pharmaceutical composition is administered sequentially, simultaneously or separately with at least one active agent.

Therapeutic method

According to a further aspect of the present invention there is provided an isolated anti-vδ1 antibody or antigen binding fragment thereof as defined herein for use as a medicament. References herein to "using" an antibody or antigen-binding fragment thereof as a medicament or in therapy are limited to administration of the antibody or antigen-binding fragment thereof to a subject.

In one embodiment, the anti-vδ1 antibody or antigen binding fragment thereof is used in the treatment of cancer, infectious disease or inflammatory disease. In one embodiment, the invention is a method of treating a disease or disorder in a subject in need thereof, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof. In various embodiments, the disease or disorder is cancer, infectious disease, or inflammatory disease. In one embodiment, the anti-vδ1 antibody or antigen binding fragment thereof is used in the treatment of cancer, infectious disease or inflammatory disease, causing diseased cells to die while retaining healthy cells. In another embodiment, the antibody or antigen binding fragment thereof is used in the treatment of cancer.

In one embodiment, the antibody or antigen binding fragment thereof is used in the treatment of cancer, infectious disease, or inflammatory disease. In another embodiment, the antibody or antigen binding fragment thereof is used in the treatment of cancer.

According to a further aspect of the present invention there is provided a pharmaceutical composition as defined herein for use as a medicament. In one embodiment, the pharmaceutical composition is used in the treatment of cancer, infectious disease or inflammatory disease. In another embodiment, the pharmaceutical composition is for use in the treatment of cancer.

According to a further aspect of the present invention there is provided a method of modulating an immune response in a subject in need thereof, the method comprising administering a therapeutically effective amount of an isolated anti-vδ1 antibody or antigen binding fragment thereof as defined herein. In various embodiments, modulating an immune response in a subject comprises binding or targeting γδ T cells, activating γδ T cells, causing or increasing γδ T cell proliferation, causing or increasing γδ T cell expansion, causing or increasing γδ T cell degranulation, causing or increasing γδ T cell mediated killing activity while retaining healthy cells, causing or increasing γδ T cytotoxicity while retaining healthy cells, causing or increasing γδ T cell mobilization, increasing γδ T cell survival, or increasing resistance to γδ T cell depletion. Modulating the immune response in the subject may also include binding or targeting a second antigen. For example, in some embodiments, the binding of the second antigen, particularly when it is an immunomodulatory antigen, may stimulate immunomodulation via the second antigen in addition to immunomodulation via binding of the multispecific antibody to TRDV 1. Thus, modulation of an immune response may include modulation via two different signaling pathways, a first signaling pathway being modulated via TRDV antigen-binding and a second signaling pathway being modulated via binding of a second immunomodulatory antigen.

According to a further aspect of the present invention there is provided a method of treating cancer, infectious disease or inflammatory disease in a subject in need thereof, the method comprising administering a therapeutically effective amount of an isolated anti-vδ1 antibody or antigen binding fragment thereof as defined herein. Alternatively, a therapeutically effective amount of the pharmaceutical composition is administered.

According to a further aspect of the invention there is provided the use of an antibody or antigen binding fragment thereof as defined herein for the manufacture of a medicament for use, for example, in the treatment of cancer, infectious disease or inflammatory disease.

In one embodiment, the antibody or antigen binding fragment thereof is administered to a subject, wherein the subject has cancer, an infectious disease, or an inflammatory disease.

According to a further aspect of the present invention there is provided a pharmaceutical composition as defined herein for use as a medicament. In one embodiment, the pharmaceutical composition is administered to a subject, wherein the subject has cancer, an infectious disease, or an inflammatory disease.

According to another aspect of the present invention there is provided a method of administering a therapeutically effective amount of an isolated anti-vδ1 antibody or antigen binding fragment thereof as defined herein to a subject, wherein said subject has cancer, an infectious disease or an inflammatory disease. Alternatively, a therapeutically effective amount of the pharmaceutical composition is administered.

According to a further aspect of the invention there is provided the use of an antibody or antigen binding fragment thereof as defined herein for the manufacture of a medicament, for example for administration to a subject suffering from cancer, infectious disease or inflammatory disease.

In various embodiments, cancers treatable by the disclosed methods and compositions include, but are not limited to: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, appendicular carcinoma, basal cell carcinoma, cholangiocarcinoma, bladder carcinoma, bone carcinoma, osteosarcoma and malignant fibrous histiocytoma, brain stem glioma, brain tumor, brain stem glioma, central nervous system atypical teratoma/rhabdomyoma, central nervous system embryonal tumor, cerebellar astrocytoma, brain astrocytoma/glioblastoma, craniopharyngeal tumor, ependymal tumor, medulloblastoma, mesogenic pineal parenchymal tumor, supracurtain primitive neuroectodermal tumor and pineal blastoma, visual pathway and hypothalamic glioma, brain and spinal cord tumor, breast carcinoma, bronchotumor, burkitt lymphoma (Burkitt lymphoma), carcinoid tumor, gastrointestinal carcinoid tumor atypical teratoma/rhabdomyoma of central nervous system, embryonic tumor of central nervous system, central nervous system lymphoma, cerebellar astrocytoma, brain astrocytoma/glioblastoma, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngeal tube tumor, cutaneous T-cell lymphoma, esophageal cancer, especially family tumor (EWING FAMILY of tumors), extragonadal germ cell tumor, extrahepatic bile duct cancer, intraocular melanoma, retinoblastoma, gallbladder cancer, gastric cancer (gastric/stomach cancer), gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (gist), germ cell tumor, gestational trophoblastoma, glioma, brain stem glioma, glioblastoma, visual pathway and hypothalamic glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, langerhans cell histiocytoma (LANGERHANS CELL histiocytosis), hodgkin's lymphoma (Hodgkin's lymphoma), hypopharyngeal cancer, hypothalamic and ocular pathway glioma, intraocular melanoma, islet cell tumor, renal (renal cell) cancer, langerhans cell histiocytoma, laryngeal cancer, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, hairy cell leukemia, lip cancer and oral cancer (oral CAVITY CANCER), liver cancer, non-small cell lung cancer, aids related lymphomas, burkitt's lymphoma, cutaneous T cell lymphoma, non-Hodgkin's lymphoma, primary central nervous system lymphoma, waldenstrom's macroglobulinemia (Waldenstrom macroglobulinemia) osteomalignant fibrous histiomas and osteosarcomas, medulloblastomas, melanomas, merkel cell neoplasms (MERKEL CELL carcinoma), mesothelioma, occult primary metastatic squamous neck cancer, oral cancer (mouth cancer), multiple endocrine tumor syndrome, multiple myeloma/plasmacytomer, mycomycosis fungoides, myelodysplastic syndrome, myelodysplastic/myeloproliferative disorders, myelogenous leukemia, myeloid leukemia, acute myeloid leukemia, multiple myeloma, myeloproliferative disorders, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastomas, non-small cell lung cancer, oral cancer (oral cancer), oral cancer, oropharyngeal cancer, osteosarcoma and osteomalignant fibrous histiocytomas, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumors, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, mesogenic pineal parenchymal tumor, pineal blastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, plasmacytoma/multiple myeloma, pleural pneumoblastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter cancer, respiratory tract cancer involving the nut gene on chromosome 15, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, ewing's family tumor, kaposi's sarcoma (Kaposi's sarcoma), tumor soft tissue sarcoma, uterine sarcoma, sezary syndrome (non-melanoma), skin cancer (melanoma), merck cell skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer, gastric cancer, supratentorial primitive neuroectodermal tumors, T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymus cancer, thyroid cancer, renal pelvis and ureter transitional cell carcinoma, gestational trophoblastoma, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, waldenstrom's macroglobulinemia and Wilms tumor (Wilms tumor). In various embodiments, cancers treatable by the disclosed methods and compositions are treated while retaining healthy cells.

In the context of a variety of embodiments of the present invention, inflammatory diseases treatable by the disclosed methods and compositions include, but are not limited to, cardiac achalasia, acute Disseminated Encephalomyelitis (ADEM), acute motor axonal neuropathy, acute Respiratory Distress Syndrome (ARDS), ai Disen's Disease, painful obesity, adult Still's Disease, adult-onset Still Disease, agaropectinemia, alopecia areata, amyloidosis, amyotrophic lateral sclerosis, ankylosing spondylitis, anti-GBM/anti-TBM nephritis, anti-N-methyl D-aspartate (anti-NMDA) receptor encephalitis, anti-phospholipid syndrome (APS, APLS), anti-synthase syndrome, anti-tubular basement membrane nephritis, aplastic anemia, atopic allergy atopic dermatitis, autoimmune angioedema, autoimmune concurrent Disease, autoimmune autonomic dysfunction, autoimmune encephalomyelitis, autoimmune enteropathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune Inner Ear Disease (AIED), autoimmune lymphoproliferative syndrome, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis (AIP), autoimmune peripheral neuropathy, autoimmune multiple endocrine syndrome (APS) type 1, autoimmune multiple endocrine syndrome (APS) type 2, autoimmune multiple endocrine syndrome (APS) type 3, autoimmune retinopathy, autoimmune thrombocytopenic purpura, autoimmune thyroiditis, autoimmune urticaria, autoimmune uveitis, autoimmune vasculitis, axonal and neuronal neuropathy (AMAN), baloc concentric sclerosis (Balo concentric sclerosis), baluo Disease (Bal Disease), behcet's Disease, benign mucosal pemphigoid, bikerstaff's encephalitis (bicerstaff 'S ENCEPHALITIS), bulaque Syndrome (Blau syncronome), bullous pemphigoid, castelman Disease (CASTLEMAN DISEASE, CD), celiac Disease, chagas Disease (CHAGAS DISEASE), chronic fatigue Syndrome, chronic Inflammatory Demyelinating Polyneuropathy (CIDP), chronic obstructive pulmonary Disease, chronic Recurrent Multifocal Osteomyelitis (CRMO), chessman Syndrome (Churg-Strauss synome, CSS) or Eosinophilic Granuloma (EGPA), cicatricial pemphigoid, cogan Syndrome, collectin, complement component 2 deficiency, complex regional pain Syndrome, congenital heart block, connective tissue, systemic and multiple organ contact dermatitis, coxsackie viral myocarditis (Coxsackie myocarditis), CREST Syndrome, crohn's Disease, cox's Syndrome (Cushing's Syndrome), cutaneous leucocyte-disrupting vasculitis, digodisease (Dego's Disease), dermatitis herpetiformis, dermatomyositis, devic's Disease (neuromyelitis), type 1 diabetes mellitus, digestive system, discoid lupus, derehler syndrome (Dressler's syndrome), drug lupus, eczema, endometriosis, arthritis associated with attachment point inflammation, eosinophilic esophagitis (EoE), eosinophilic fasciitis, eosinophilic gastroenteritis, eosinophilic granulomatosis with polyangiitis (EGPA), eosinophilic pneumonia, acquired epidermolysis bullosa, nodular erythema, fetal erythropoiesis, esophageal achalasia, primary mixed cryoglobulinemia, evans syndrome (Evans syndrome), exocrine, fisher's syndrome (Feltey syndrome), progressive fibrodysplasia, fibromyalgia, fibroalveolar inflammation, gastritis, gastrointestinal pemphigoid, giant cell arteritis (temporal arteritis), giant cell myositis, glomerulonephritis, gopasmodus syndrome (Gopasmodus's syndrome), goprandial syndrome (Goldsupport's), graves's syndrome, graves-Schonlein-Schleman syndrome (Graves's disease), graves-Schoendo's disease, graves-Schleme ' disease (Graves ' disease), graves-Schoendo's disease (Graves-Schoen ' disease), graves-Graves ' disease (Graves ' disease, HSP), herpes or pemphigoid of Pregnancy (PG), hidradenitis Suppurativa (HS) (acne), hypogammaglobulinemia, idiopathic giant cell myocarditis, idiopathic inflammatory demyelinating diseases, idiopathic pulmonary demyelinating diseases, igA nephropathy, igA vasculitis (IgAV), igA vasculitis, and, igG 4-related diseases, igG 4-related sclerotic diseases, immune Thrombocytopenic Purpura (ITP), inclusion Body Myositis (IBM), inflammatory bowel disease, moderate uveitis, interstitial Cystitis (IC), interstitial lung disease, IPEX syndrome, juvenile arthritis, juvenile diabetes (type 1 diabetes), juvenile Myositis (JM), kawasaki disease (KAWASAKI DISEASE), lanbert-Eaton syndrome (Lambert-Eaton syndrome), white blood cell disruption vasculitis, lichen planus, lichen sclerosus, linear IgA disease (LAD), lupus nephritis, lupus vasculitis, chronic lyme disease (LYME DISEASE chronic), ma Jide syndrome (Majeed syndrome), meniere's disease Microscopic Polyangiitis (MPA), mixed Connective Tissue Disease (MCTD), mo Lunshi ulcers (Mooren's ulcer), scleroderma, mulbergial-Hash disease (Mucha-Habermann disease), multifocal Motor Neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myocarditis, myositis, somnolence, neonatal lupus, nervous system neuromyelitis, neuromyotonia, neutropenia, ocular scarring pemphigoid, strabismus-eye clonus myoclonus syndrome, optic neuritis, ord's thyroiditis, field primary syndrome (Oshtoran syndrome), recurrent rheumatism (PR), paraneoplastic Cerebropathyosis (PCD), paroxysmal Nocturnal Hemoglobinuria (PNH), paris-Long Bage syndrome (Parry-Romberg syndrome), parsen-Tener syndrome (Parsonage-Turner syndrome), pars plana (peripheral uveitis), streptococcus-related childhood autoimmune neuropsychiatric disorder (PANDAS), pelvic Inflammatory Disease (PID), pemphigus vulgaris, peripheral neuropathy, perivenous encephalomyelitis, pernicious Anemia (PA), acute acne-like lichen-like pityriasis, MS syndrome, polyarteritis nodosa, type I, II, III polyadenylic syndrome, polymyalgia rheumatica, polymyositis, post myocardial infarction syndrome, pericardial post-operative syndrome, primary Biliary Cholangitis (PBC), primary biliary cirrhosis primary immunodeficiency, primary sclerosing cholangitis, progesterone dermatitis, progressive inflammatory neuropathy, psoriasis, psoriatic arthritis, pure red blood cell dysgenesis (PRCA), pyoderma gangrenosum, las Mu Sen encephalitis (Rasmussen 'S ENCEPHALITIS), raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, recurrent polychondritis, restless Leg Syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, rheumatoid vasculitis, sarcoidosis, schizophrenia, schmidt syndrome (Schmidt syndrome), schmidt syndrome (Schnitzler syndrome), scleritis, scleroderma, serous disease, sjogren syndrome)Syndrome), sperm and testis autoimmunity, spondyloarthropathies, stiff Person Syndrome (SPS), subacute Bacterial Endocarditis (SBE), sulsaxox Syndrome (Susac's Syndrome), swatt's Syndrome (Sweet's Syndrome), cidenham's chorea, sympathogenic Ophthalmitis (SO), systemic Lupus Erythematosus (SLE), takayasu ' S ARTERITIS, temporal arteritis/giant cell arteritis, thrombocytopenia, thrombocytopenic purpura (TTP), thyroiditis, tolosa-hunter Syndrome (Tolosa-Hunt Syndrome) (THS), transverse myelitis, type 1 diabetes mellitus, ulcerative Colitis (UC), undifferentiated Connective Tissue Disease (UCTD), undifferentiated spondyloarthropathies, urticaria vasculitis, urticaria, uveitis, vasculitis, vitiligo, and voglio-small-primordial Tian Bing (kogaog-49498-4). In various embodiments, inflammatory diseases treatable by the disclosed methods and compositions are treated while preserving healthy cells.

In various embodiments, infectious diseases treatable by the disclosed methods and compositions include, but are not limited to, acinetobacter infection, actinomycosis, acute relaxant myelitis (AFM), african comatose (african trypanosomiasis), AIDS (acquired immunodeficiency syndrome), amoeba infection, amoeba disease, phagostimulant infection, anaplasmosis, angiostrongylosis, alien-tip nematode disease, anthrax, neuroinvasive and non-neuroinvasive arbovirus disease, haemolytic secretion bacterial infection, argentina hemorrhagic fever, ascariasis, aspergillosis, astrovirus infection, avian influenza, babesia, bacillus cereus infection, bacterial meningitis, bacterial pneumonia, bacterial vaginosis, bacteroides infection, ciliate disease, baltosomiasis, bezoa infection (Baylisascaris infection), BK virus infection, sarcoidosis, blastosis, botulism (food-borne), botulism (botulism), botulism (infant) infection, bruxism (bruxism), bruxism (calixazosis), bruxism (calixagliosis), bruxism (calixazosis), bruxism (32) infection with blood-fluid-borne by other bacteria, bruxism (calixazosis), bruxism (calixagliosis) infection, bruxism) and other infections (calixanica virus (32) infection, bruxism) infection with calixaco bacteria, bruxism (calibrio virus, bruxism) infection, bruxism-induced by calix, and bruxism infection; thrush) a part, capillary Disease, carbapenemase-producing bacteria of the family enterobacteriaceae (CP-CRE), carbapenemase-resistant infections (CRE/CRPA), bartonella Disease, cat scratch Disease, cellulitis, chagas Disease (trypanosomiasis), chancroid, varicella, chikungunya virus infection (Chikungunya Virus Infection) (chikungunya fever), chlamydia trachomatis, chlamydia pneumoniae infection, cholera, yellow parent Disease, pot Disease, cigazemia (Ciguatera), clonorchiasis, clostridium difficile colitis, clostridium difficile infection, clostridium perfringens infection, coccidioidomycosis infection (cereal fever), corradon Tick Fever (CTF), common cold (acute viral nasopharyngitis; acute rhinitis), congenital syphilis, conjunctivitis, COVID-19 (2019 coronavirus Disease), enterobacter CP-CRE, escherichia coli (escherichia coli) CP-CRE, klebsiella CP-CRE, creutzfeldt-Jacob Disease, transmissible spongiform encephalopathy (CJD), creutzfeldt-jakob Disease (CCHF), scab, cryptococcosis, cryptosporidiosis (Cryptoo), skin larva migration disorder (CLM), cyclosporinosis, cysticercosis, cytomegalovirus infection, dengue virus infection, dengue 1,2, 3, 4 (dengue), dengue-like Disease, chain belt infection (Desmodesmus infection), diarrhea Disease, binuclear amebiasis (Dientamoebiasis), cryptosporidiosis (CLM), cyclosporosis (CLM), and the like, diphtheria, cestodiasis, mule, colibacillosis, shigellosis (STEC), eastern equine encephalitis virus, ebola hemorrhagic fever (ebola), echinococcosis, chafevere, eichorea infection (EHRLICHIAEWINGII INFECTION), eichorea, apoplast, encephalitis, arbovirus or side infection, enterobiasis (enterobiasis infection), enterococcidiosis, enterovirus infection, enterovirus D68 (EV-D68) infection, non-poliomyelitis (non-polio enterovirus), epidemic typhus, epstein-barr virus infectious mononucleosis (Epstein-Barrvirus infectious mononucleosis) (Mono), infectious erythema (fifth disease), bursa (sixth disease), fasciolopsis, gingiva (Fasciolopsiasis), fatal Familial Insomnia (FFI), wuchorea, influenza (seasonal), food poisoning, non-established parasitic infection, clostridia, fungus, gella, genital infection, gella, genital herpes, genital infections, gella, and so-quail's disease (german-genital syndrome)Scheinkersyndrome) (GSS), giardiasis, jaundice, jaw nematode, gonorrhea, inguinal granuloma (Du Nuofan mycosis (Donovanosis)), group A streptococcal infection, group A streptococci, group B streptococcal infection, melon-and-support virus infection, haemophilus influenzae, type B (Hib or H influenza), haemophilus influenzae infection, hand-foot-and-mouth Disease (HFMD), hansen's Disease, hantaan virus infection (Hantavirus infection), hantaan virus lung syndrome (HPS), abdominal ground virus Disease, helicobacter pylori infection, hemolytic Uremic Syndrome (HUS), hemorrhagic fever with renal syndrome (HFRS), hendela virus infection (Hendra virus infection) hepatitis A (Hep A), hepatitis B (Hep B), hepatitis C (Hep C), hepatitis D (Hep D), hepatitis E (Hep E), herpes B, herpes simplex, herpes zoster VZV (shingles), haemophilus influenzae type B (Hib Disease), histoplasmosis (histoplasmosis), hookworm infection, HPV (human papilloma virus), human Bocka virus infection, human Ehrlich's Disease (Human ewingii ehrlichiosis), human granulocyte-colony anaplasma Disease (HGA), human immunodeficiency virus/AIDS (HIV/AIDS), human metapneumovirus infection, human monocyte Ehrlich's Disease, human Papilloma Virus (HPV) infection, human parainfluenza virus infection, membrane shell taeniasis, impetigo, influenza (flu), influenza (seasonal), invasive pneumococcal disease, isospora, hooning virus infection, kawasaki syndrome (Kawasaki Syndrome), keratitis, gold infection (KINGELLA KINGAE in), kuru, lassa fever (LASSA FEVER), lassa virus infection, legionellosis (Legionella), leishmaniasis, leprosy (Hansen's disease), leptospirosis, listeriosis (listeria), ruabout virus (Lujo virus) infection, lyme disease (LYME DISEASE), lymphofilariasis (elephant skin disease), lymphocytic Choriomeningitis (LCMV), venereal lymphogranuloma infection (LGV), ma Qiubo virus infection, malaria, leprosy Marburg virus infection, measles, melioidosis (Whitermor's disease), meningitis, bacterial meningitis, viral meningitis, meningococcal disease, postschistosomiasis, microsporidian disease, middle East Respiratory Syndrome (MERS), molluscum Contagiosum (MC), monkey pox, mononucleosis, mosquito-borne disease, MRSA, mumps, murine typhus (endemic typhus), podophyllosis, mycoplasma genitalia infection, mycoplasma pneumonia, myiasis, neisseria meningitidis infection, neonatal conjunctivitis (neonatal ophthalmitis), nipah virus infection, nocardia, norovirus infection, disc tail filariasis (river blindness), posttestosterone fluke disease, orf virus infection (orodynia), pycnopsis (Nanmeric blastosis), pneumochiosis, paralytic shellfish poisoning (paralytic shellfish poisoning, siga poisoning), pasteuresis, PEP, parasitic infection, pertussis (Pertussis/whooping cough), pinkeye, pneumococcal infection, pneumocystis pneumonia (PCP), pneumonia, plague, poliomyelitis (Polio), paralytic poliomyelitis, poliomyelitis virus infection, pang Dike fever, powa mulberry virus disease (Powassan virus disease), prevotella infection, primary amoebic encephalitis (PAM), progressive multifocal leukoencephalopathy, protozoal infection, psittacosis (psittacosis), pustular rash (smallpox, monkey pox, vaccinia), rabies raccoon roundworm infection, mouse bite fever, pool disease (Recreational WATER ILLNESSES), regression fever, respiratory syncytial virus infection, reye's Syndrome (Reye's syncrome), rhinosporidiosis, rhinovirus infection, rickettsia (hizipras fever), rift Valley Fever (RVF), ringworm, rotavirus infection, rubella, sha Biya virus infection, salmonella paratyphi infection, salmonella typhi infection, salmonellosis, SARS (severe acute respiratory Syndrome), scabies, scarlet fever, schistosomiasis, mackerel poisoning (Scombroid), sepsis, septic shock, septicemia, severe Acute Respiratory Syndrome (SARS), shiga toxin-producing escherichia coli, shigella infection, shigellosis, herpes (shingles), smallpox, mouth pain (Orf virus), sporozoosis, spot fever rickettsia, st.louis ENCEPHALITIS VIRUS DISEASE, staphylococcal infection (methicillin resistant (MRSA)), staphylococcal food poisoning, staphylococcal infection (vancomycin intermediate (VISA)), septic pharyngolaryngitis, group a streptococcal disease (invasive), group B streptococcal disease (Strep-B), streptococcal toxic shock syndrome, round-wire-like disease, subacute sclerosing encephalitis, syphilis, taeniasis, tetanus infection, tick-borne disease, tinea capitis tinea corporis, tinea cruris, tinea manuum, tinea nigra, tinea pedis, onychomycosis, tinea versicolor, toxic shock syndrome, toxoplasmosis (ocular larval transfer (OLM)), toxoplasmosis (visceral larval transfer (VLM)), toxoplasmosis, trachoma, trichomatosis, trichomoniasis, hair disease infection (trichinosis), whipdisease (whipworm infection), tuberculosis (TB), rabbit fever (Tularemia/Rabbit fever), typhoid fever, group D typhoid fever, typhus fever, mycoplasma urealyticum infection, colpitis, brook fever, variant type creutzfeldt-jakob disease (vCJD, nvd), varicella (Varicella/Chickenpox), venezuelan equine encephalitis, venezuelan hemorrhagic fever, vibrio cholerae infection (cholera), vibrio parahaemolyticus enteritis, vibrio vulnificus infection, vibriosis, viral infection, viral hemorrhagic fever (ebola, lassa fever, marburg disease), viral Hemorrhagic Fever (VHF), viral pneumonia, west nile virus disease, west equine encephalitis virus disease, white hair knot disease (tinea alba), pertussis, yellow fever, yersinia disease (yersinia), yersinia pseudotuberculosis infection, yersinia disease, zhai ka disease (Zeaspora), zhai ka fever, zhai ka virus disease, congenital zhai ka virus disease, non-congenital zhai ka virus disease, zhai ka virus infection (zhai ka virus), congenital zhai ka virus infection, non-congenital zhai ka virus infection, and zygomycosis. In various embodiments, infectious diseases treatable by the disclosed methods and compositions are treated while preserving healthy cells.

In one embodiment, the invention is a method of activating at least one γδ T cell in a subject, the method comprising the step of administering an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of inducing or increasing γδ T cell proliferation in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of causing or increasing γδ T cell degranulation in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of eliciting or increasing γδ T cell killing activity (e.g., T cell mediated killing activity) in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein. In one embodiment, the invention is a method of eliciting or increasing γδ T cell killing activity (e.g., T cell mediated killing activity) while retaining healthy cells in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of eliciting or increasing γδ T cytotoxicity in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein. In one embodiment, the invention is a method of eliciting or increasing γδ T cytotoxicity while retaining healthy cells in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of causing or increasing γδ T cell mobilization in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of increasing γδ T cell survival in a subject, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

In one embodiment, the invention is a method of increasing the resistance of a subject to γδ T cell depletion, the method comprising the step of administering to the subject an anti-vδ1 antibody or antigen binding fragment thereof as defined herein.

According to another aspect of the invention, there is provided a method of stimulating an immune response in a subject, the method comprising administering to the subject an amount of an anti-vδ1 antibody or antigen binding fragment thereof effective to stimulate an immune response.

Use of antibodies or antigen binding fragments thereof

According to another aspect of the invention there is provided the use of an anti-vδ1 antibody or antigen binding fragment thereof as described herein for studying antigen recognition, activation, signal transduction or function of γδ T cells, in particular vδ1T cells. As described herein, the antibodies have been demonstrated to be active in assays useful for exploring γδ T cell function. Such antibodies may also be used to induce proliferation of γδ T cells and thus may be used in methods of expanding γδ T cells (e.g., vδ1T cells).

Antibodies that bind to the vδ1 chain may be used to detect γδ T cells. For example, antibodies can be labeled with a detectable label or reporter or used as a capture ligand to selectively detect and/or isolate vδ1T cells in a sample. The labeled antibodies can be used in a number of ways known in the art, such as immunohistochemistry and ELISA.

The detectable label or reporter may be a radioisotope, such as ³H、¹⁴C、³²P、³⁵ S or ¹²⁵ 1; fluorescent or chemiluminescent moieties, such as fluorescein isothiocyanate or rhodamine; or an enzyme such as alkaline phosphatase, beta-galactosidase, horseradish peroxidase or luciferase. The fluorescent labels applied to the antibodies of the invention can then be used in Fluorescence Activated Cell Sorting (FACS) methods.

Thus, in various embodiments, the invention includes an in vivo method of modulating γδ T cells, a method of binding γδ T cells, a method of targeting γδ T cells, a method of activating γδ T cells, a method of proliferating γδ T cells, a method of expanding γδ T cells, a method of detecting γδ T cells, a method of causing γδ T cell degranulation, a method of causing γδ T cell mediated killing activity, a method of selecting an antibody or antigen binding fragment thereof, the method comprising the step of administering an anti- γδ antibody or antigen binding fragment thereof as described herein to a subject.

Drugs for modulating γδ T cells

The anti-vδ1 antibodies or antigen binding fragments thereof as described herein may be used for in situ (i.e. in vivo) modulation or may be used to modulate delta variable 1 chain (vδ1) T cells in a patient. The antibody or antigen binding fragment thereof may be included in a medicament for achieving such a purpose.

Modulating vδ1T cells may include:

Expansion of vδ1T cells, for example by selectively increasing the number of vδ1T cells or promoting survival of vδ1T cells;

stimulation of vδ1T cells, for example by increasing vδ1T cell potency, i.e. increasing target cell killing;

Prevention of vδ1T cell depletion, for example by increasing the persistence of vδ1T cells;

-degranulation of vδ1T cells;

-increasing NCR expression;

-increasing 4-1BB expression;

Immunoregulatory vδ1T cells, for example by down-regulating vδ1 TCR cell surface expression, i.e. by causing vδ1 TCR internalization or reducing expression of vδ1 TCR protein, or blocking vδ1 TCR binding; and/or

Down-regulating the TCR/CD3 complex.

Unlike prior art anti-vδ1 antibodies that focused on vδ1T cell depletion, the antibodies of the present invention may be used to activate vδ1T cells via the TRDV1 binding domain. Although they may lead to down-regulation of TCR on T cells to which they bind, they do not lead to V delta 1T cell depletion, but rather they may stimulate T cells and thus may be used in a therapeutic setting that would benefit from activation of this T cell compartment. Activation of vδ1T cells was evident by TCR down-regulation, CD3 down-regulation, changes in activation markers such as CD25 and Ki67 and degranulation marker CD107 a. Activation of vδ1T cells in turn leads to release of inflammatory cytokines such as infγ and tnfα, thereby promoting immune licensing.

Drugs that modulate immune cell markers on vδ1+ cells

The antibody or antigen binding fragment thereof may modulate the immune cell markers of vδ1+ cells when administered to a patient.

The antibodies or antigen binding fragments thereof described herein may also be evaluated for suitability for therapeutic use by measuring γδt modulation. For example, by measuring the change in the level of CD25 or CD69 or CD107a present on vδ1+ T cells in the model system. Such markers are typically used as markers for lymphocyte regulation (e.g., proliferation or degranulation) and can be measured, for example, by flow cytometry after application of an antibody or antigen-binding fragment thereof as described herein. Surprisingly, during such an assessment (see, e.g., examples 7, 17, 18), antibodies as described herein were observed to confer measureable higher levels of CD25 or CD69 or CD107a levels on target vδ1+ T cells. Optionally, the phenotypic change of the vδ1+ cells or population thereof tested in the model system may then be compared to the phenotypic change when an alternative comparison antibody (e.g. OKT-3, TS8.2, etc.) is applied to the equivalent γδt cells.

Thus, in one aspect of the invention there is provided a method of assessing antibodies or antigen binding fragments thereof that bind to the vδ1 chain of γδ TCR for therapeutic use, the method comprising administering the antibodies or antigen binding fragments thereof to a population of cells comprising vδ1+ cells and determining the effect on CD25 and/or CD69 and/or CD107a levels on the surface of the vδ1+ cells. The effect on CD25, CD69 and/or CD107a levels may be determined/measured over a period of time. It will be appreciated that the effect can be measured by comparison with the level of CD25 and/or CD69 and/or CD107a on the surface of vδ1+ cells over the same period of time when the antibody is not applied to the cells. In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by adding the antibody to a population of cells comprising vδ1+ cells and then measuring the level (or expression) of CD25 or CD69 or CD107a on the surface of the vδ1+ cells.

Agents that modulate the growth characteristics or number of vδ1+ cells

The antibody or antigen binding fragment thereof may modulate the growth characteristics of vδ1+ cells after administration to a patient. For example, the antibody or antigen binding fragment thereof may expand vδ1+ cells.

An alternative method of measuring γδt proliferation may comprise measuring the relative amount of vδ1+ cells as the antibodies or antigen binding fragments thereof as described herein are applied to a model system containing said cells over time. Surprisingly, during such an assessment, it was observed that the antibodies as described herein can measurably increase the number of vδ1+ T cells (see e.g. examples 10, 17 and 18), optionally then such a change in number can be compared to the change in number observed when a surrogate comparison antibody (e.g. anti-OKT 3) is applied to the model system.

Thus, in another aspect of the invention, there is provided a method of assessing antibodies or antigen binding fragments thereof that bind to the vδ1 chain of a γδ TCR, the method comprising administering the antibodies or antigen binding fragments thereof to a population of cells comprising vδ1+ cells and determining the effect on the number of vδ1+ cells in the population. The effect on cell number can be determined/measured over a period of time. It will be appreciated that the effect can be measured in comparison to the effect on cell number observed over the same period of time when the antibody is not applied to the cell population. In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by applying the antibody to a population of cells comprising vδ1+ cells, and then measuring the number of said cells over time.

Agents that modulate the proliferative capacity and number of vδ1+ cells

The ideal therapeutic antibody or antigen binding fragment thereof that binds to the vδ1 chain of γδ TCR as described herein may be an antibody or antigen binding fragment thereof capable of enhancing vδ1+ cell proliferation in vivo. Such antibodies can then be used as agents designed to specifically increase the number of vδ1+ cells in a subject or patient. For example:

Cancer:

The relative increase in the number of V.delta.1+ cells is reported to be a positive prognostic indicator associated with the improvement of prognosis for many cancers (see, e.g., gentles et al (2015) Nature Immunology 21:938-945; wu et al (2019) Sci. Trans. Med.11 (513): eaax9364; CATELLANI et al (2007) Blood 109 (5): 2078-2085). In one embodiment, presented herein is a drug capable of increasing the relative or absolute number of vδ1+ cells in situ in a cancer patient.

Pathogenic/parasitic/viral infection:

During host defense against various acquired pathogenic/parasitic/viral infections, vδ1+ cell enrichment was observed. For a recent general review, see Zhao et al (2018) immunol. Res.2018:5081634. in addition, the increased number of vδ1+ is also believed to prevent various DNA and RNA viral infections. For example, increased numbers are also believed to have protective effects during CMV infection associated with allograft (see van Dorp et al (2011) Biology of Blood and Marrow Transplantation (2): S217). In addition, the number of V.delta. + cells in coronavirus infected patients is increased (Poccia et al (2006) J.Infect. Dis.193 (9): 1244-1249).

In another embodiment, presented herein are agents capable of increasing the relative or absolute number of vδ1+ cells in a subject or patient carrying a pathogenic infection.

Stem cell transplantation:

An increase in the number of V.delta.1+ cells during hematopoietic stem cell transplantation has also been associated with reduced disease recurrence, fewer viral infections, increased overall survival and disease-free survival, and generally favorable clinical outcomes (see, e.g., aruda et al (2019) Blood 3 (21): 3436-3448 and Godder et al (2007) Bone Marrow Transplantation 39:751-757). Thus, another embodiment presented herein is a drug capable of increasing the relative or absolute number of vδ1+ cells in a subject as part of a therapeutic regimen that supports stem cell transplantation.

Therefore, there is a great need for a drug that is capable of preferentially or specifically increasing the number of vδ1+ cells in situ.

Agents that maintain or induce or increase cytokine secretion by vδ1+ cells

Cytokines are a large class of proteins, peptides or glycoproteins secreted by specific cells of the immune system. They are a class of signaling molecules that mediate and regulate immune, inflammatory and hematopoietic functions. Many cytokines are involved in improving the signs and symptoms of disease by directly or indirectly modulating the tumor and cellular microenvironment, autoimmune tissues and related microenvironment or virally infected tissues or cellular environments. Exemplary pro-inflammatory cytokines include tumor necrosis factor-alpha (tnfα) and interferon-gamma (ifnγ).

However, many of these cytokines exhibit adverse toxicity when administered systemically. For example, while tnfα can induce hemorrhagic necrosis of transplanted tumors and is reported to exert synergistic anti-tumor effects when combined with other chemotherapeutic agents, various clinical trials of systemic recombinant human tnfα (rhtnfα) have highlighted significant dose limiting side effects including hypotension, chills, phlebitis, thrombocytopenia, leukopenia and hepatotoxicity, fever, fatigue, nausea/vomiting, discomfort and weakness, headache, chest distress, lumbago, diarrhea and shortness of breath.

Similar systemic toxicity challenges are faced with the use of recombinant ifnγ. For example, adverse side effects are also observed, although ifnγ can exert beneficial pleiotropic effects in the cancer setting, including upregulation of MHC class I and II to stimulate anti-tumor immunity, increase T cell infiltration, confer anti-angiogenic effects, induce chemokine/cytokine secretion, and exert direct cancer cell antiproliferative effects. These side effects include fever, headache, chills, fatigue, diarrhea, nausea, vomiting, anorexia, temporary elevation of hepatic transaminase, and temporary decrease of granulocyte and leukocyte counts.

For a recent review of the potential and limitations of systemic recombinant tnfα and ifnγ, see Shen et al (2018) Cell prolif.51 (4): e12441.

Thus, there is a need for more controlled in situ, more localized, more tissue or cell specific production of such cytokines. For example, more controlled expression or induction of pro-inflammatory cytokines is proposed as a means to make a "cold" tumor convertible into a "hot" tumor. Thermal tumors are sometimes also referred to as "T cell inflammatory" because an increase in the number or density of cd45+ T cells is also observed. For a recent review, see Bonaventura et al (2019) front. Immunol.10:168.

For such reasons, a desirable therapeutic antibody or antigen-binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein may be an antibody or antigen-binding fragment thereof that can maintain or enhance or induce cytokine secretion in vδ1+ cells in vivo. Such antibodies can then be used as drugs designed to specifically increase or induce cytokines in a more localized, less systemic manner in a subject or patient, and which better correlate with the distribution of vδ1+ cells in the subject or patient.

Notably, significantly higher levels of secreted cytokines were observed when antibodies that bind to the vδ1 chain of γδ TCR as described herein were applied to vδ1+ cells. More specifically, and as a non-limiting example, significantly higher levels of tnfα and ifnγ were observed. See, e.g., example 15.

Thus, in another aspect of the invention, there is provided a method of assessing antibodies or antigen-binding fragments thereof that bind to the vδ1 chain of a γδ TCR, the method comprising administering the antibodies or antigen-binding fragments thereof to a population of cells comprising vδ1+ cells and determining the amount of at least one cytokine produced by the population of cells. The amount of cytokine produced can be determined/measured over a period of time and optionally compared to the amount observed over the same period of time when the antibody is not applied to the cell population. In one embodiment, the level of cytokine produced observed when the antibody is administered to a population of cells is greater than about 10%, greater than about 20%, greater than about 30%, greater than about 50%, greater than about 100%, greater than about 150%, greater than about 200%, greater than about 250%, greater than about 300%, greater than about 350%, greater than about 400%, greater than about 450%, greater than about 500%, greater than about 1000% relative to the level of cytokine produced when the antibody is not administered. In another aspect of the invention, the cytokine is a pro-inflammatory cytokine. In another aspect of the invention, the cytokine is a TNF- α cytokine. In another aspect of the invention is an IFN-gamma cytokine.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to a vδ1 chain of a γδ TCR as described herein by applying the antibody to a population of cells comprising vδ1+ cells and then measuring the level of at least one cytokine produced. In another aspect of the invention, the cytokine measured is a TNF- α cytokine and/or an IFN- γ cytokine.

In another aspect of the invention, there is provided a method of assessing antibodies or antigen binding fragments thereof that bind to the vδ1 chain of γδ TCR by applying the antibodies or antigen binding fragments thereof to a population of cells comprising vδ1+ cells and measuring the effect of the antibodies on modulating a colder or cold tumor into a hotter or hot tumor by determining the amount of pro-inflammatory cytokines produced and/or the number or density of cd45+ T cells present in the tumor or tumor microenvironment.

Agents for maintaining or inducing or increasing vδ1+ cell granzyme B activity

Granzyme B is a serine protease commonly found in granules of natural killer cells (NK cells) and cytotoxic T cells. It is secreted by these cells along with the pore-forming protein perforin to mediate apoptosis of target cells (e.g., diseased cells).

When vδ1+ cells are incubated with target diseased cells (e.g. cancer cells) in a co-culture in a model system, the level of granzyme B and the level of activity in the target diseased cells can be measured prior to lysis. Notably, when antibodies or antigen binding fragments thereof that bind to the vδ1 chain of γδ TCRs as described herein are subsequently applied to such co-cultures of vδ1+ cells and cancer cells in such model systems, higher levels and activity of granzyme B are subsequently observed in diseased cancer cells prior to cell death (see, e.g., example 16).

Thus, in another aspect of the invention, there is provided a method for assessing an antibody or antigen-binding fragment thereof that binds to a vδ1 chain of a γδ TCR, the method comprising administering the antibody or antigen-binding fragment thereof to a co-culture comprising vδ1+ cells and diseased cells (e.g. cancer cells) and measuring the effect on the amount of granzyme B produced by the diseased cells in the co-culture. The amount of cytokine produced can be determined/measured over a period of time and optionally compared to the amount observed over the same period of time when the antibody is not applied to the co-culture. In one embodiment, the level of granzyme B measured when the antibody is applied to the co-culture is greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 70%, greater than about 80%, greater than about 90%, greater than about 100%, greater than about 200% relative to the level of granzyme B observed when the antibody is not applied.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by applying the antibody to a co-culture comprising vδ1+ cells and diseased cells, and then measuring the amount and activity of granzyme B in the diseased cells.

Drugs that specifically up-regulate 4-1BB (CD 137) expression on V.delta.1 ⁺ T cells

4-1BB (CD 137) is a member of the TNFR superfamily and is an inducible T cell costimulatory molecule. With few exceptions, the expression of 4-1BB is activation dependent. In general, 4-1BB is not readily detectable on conventional (. Alpha.beta.) resting T cells or related T cell lines. However, 4-1BB is stably upregulated when traditional T cells are activated by a variety of agonists, such as plate-binding anti-CD 3, canavalin A, phytohemagglutinin, interleukins (IL) -2, IL-4 and CD28, as well as phorbol myristate acetate and ionomycin, used alone or in combination with APC. However, while such agents are known to generally up-regulate T cells, there remains a need for a less-slenderness, more targeted approach to avoiding the overstimulation typically conferred by anti-CD 3 drugs and the like. Furthermore, while the role and function of 4-1BB in 'traditional' αβ T cells has been well studied, its role in γδ T cells has been rarely explored. However, lee in at least one report in 2013 (https:// doi. Org/10.1002/eji. 201242842) demonstrated that 4-1BB was not normally expressed on human γδT cells (subtype not fully defined, possibly predominantly Vγ9Vδ2) resting in human peripheral blood, but that 4-1BB upregulation was achieved in the presence of IPP/IL-2 cytokines for 7 days. Furthermore, the authors subsequently demonstrated that the γδ T cells thus produced were more effective in preventing listeria infection in the housed mouse model. Recently, park and Lee 2021 (https:// doi.org/10.1038/s 12276-021-00576-0) agreed with these observations, at least for the peripheral blood dominant cell types, where they indicated that 4-1BB enhanced Vγ9Vδ2T cells (while also acknowledging that their role and mechanism is still unclear). Thus, there is a need for agents that specifically enhance 4-1BB expression on γδ T cells to treat a disease or disorder, thereby ameliorating at least one sign or symptom of the disease or disorder. This is especially true in situ, and as discussed elsewhere herein, such cells include tissue resident vδ1+ T cells that are generally considered to be 'resting'. Thus, in another embodiment, presented herein is a class of high affinity human anti-vδ1 antibodies capable of significantly increasing the level of 4-1BB in vδ1+ T cells (see fig. 64). And in yet another embodiment, presented herein is a class of high affinity human anti-vδ1 antibodies capable of significantly increasing the level of 4-1BB in tissue-derived vδ1+ T cells. For example, by common one-way analysis of variance with Sidak post-hoc testing, a significant increase may be p < 0.05.

Drugs that upregulate expression of the Native Cytotoxic Receptor (NCR) on V.delta.1 ⁺ T cells

As discussed elsewhere herein, the desired agent may be an agent that specifically targets and activates the most desired subset of T cells; in particular, it is a drug that targets and activates vδ1+ T cells. Given that such vδ1+ T cells are able to recognize and distinguish 'self' characteristics of stress or disorder (such as those found on infected or cancerous cells) by means of sensing mechanisms including engagement by Natural Cytotoxic Receptors (NCR), then the ideal drug might be one that is able to up-regulate NCR on vδ1+ T cells. In the studies presented herein (see fig. 65), all affinity matured anti-vδ1+ antibodies studied confer upregulation of vδ1+ T cell NCR in an expanded PBMC culture model.

Drug for amplifying polyclonal V delta 1+ cell population

The ideal antibody drug may also be one designed to ensure that the amplified vδ1+ cells do not become too cloned concentrated at the level of the hypervariable CDR3 sequences. Thus, ideal antibody drugs can be designed to avoid inducing vδ1+ cell proliferation by binding to the paratope of a specific or 'private' δ1+ cdr3 sequence. In contrast, antibodies can bind via conserved germline sequences present on all vδ1+ T cell receptors and in a gamma chain independent manner, rather than binding to sequences present only on a subset of vδ1+ cells.

Thus, an ideal antibody drug would stimulate vδ1+ cell expansion to generate a plurality of vδ1+ cells containing a mixture of CDR3 sequences. This in turn results in a heterogeneous polyclonal vδ1+ cell population that is expanded in vivo displaying different CDR3 sequences on the δ variable 1 chain. Notably, during analysis of the expanded vδ1+ cell population generated by the method of adding antibodies or antigen binding fragments thereof as described herein to the starting immune cell population containing vδ1+ cells, extensive polyclonality was observed by an RNAseq-based method designed to sequence through CDR3 hypervariable regions of the extracted RNA (see e.g. example 10).

Thus, in one aspect, there is provided a method of assessing antibodies or antigen-binding fragments thereof that bind to the vδ1 chain of a γδtcr, the method comprising administering the antibodies or antigen-binding fragments thereof to a population of cells comprising vδ1+ cells and determining the polyclonality of the expanded vδ1+ cells. Antibody drugs require the generation of amplified polyclonal populations containing multiple vδ1+cdr3 sequences. The polyclonality can be determined using methods known in the art, for example, by nucleic acid sequencing methods that are capable of analyzing the vδ1 chain hypervariable CDR3 content of the vδ1+ cells.

Medicine for amplifying polyclonal V delta 1+ cells for long time

An ideal antibody drug might be able to enhance or promote or stimulate proliferation of primary vδ1+ cells without depleting such cells in vivo. For example, and by comparison, anti-CD 3 drugs such as OKT3 (e.g., moromiab (Muromonab)) while able to expand CD3 positive T cells, may also deplete or induce anergy. To assess the ability of antibodies as described herein and that bind to the vδ1 chain of γδ TCRs to drive viable vδ1+ cells to sustain cell division, longer term proliferation studies were performed. Notably, these studies reveal that antibodies as described herein that bind to the vδ1 chain of γδ TCRs are able to drive cell division/proliferation of vδ1+ cells that are viable and still functionally cytotoxic for more than 40 days (see e.g. example 10).

In one embodiment, a method of evaluating an antibody or antigen-binding fragment thereof that binds to a vδ1 chain of a γδ TCR is provided, the method comprising applying the antibody or antigen-binding fragment thereof to a population of cells and monitoring the length of time that vδ1+ cell division occurs. Ideally, the antibody is capable of stimulating vδ1+ cell division for 5 to 60 days, for example at least 7 to 45 days, 7 to 21 days or 7 to 18 days.

In another embodiment, there is provided an antibody or antigen binding fragment thereof as described herein that binds to the vδ1 chain of γδ TCR and is capable of stimulating vδ1+ cell division upon administration to a patient, increasing the number by at least 2-fold, increasing the number by at least 5-fold, increasing the number by at least 10-fold, increasing the number by at least 25-fold, increasing the number by at least 50-fold, increasing the number by at least 60-fold, increasing the number by at least 70-fold, increasing the number by at least 80-fold, increasing the number by at least 90-fold, increasing the number by at least 100-fold, increasing the number by at least 200-fold, increasing the number by at least 300-fold, increasing the number by at least 400-fold, increasing the number by at least 500-fold, increasing the number by at least 600-fold, increasing the number by at least 1,000-fold.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to a vδ1 chain of a γδ TCR as described herein by applying the antibody to a vδ1+ cell population or mixed cell population containing vδ1+ cells, and then measuring the number of vδ1+ cells over time.

Drugs that modulate non-vδ1+ immune cells by targeting vδ1+ immune cells

Antibodies or antigen binding fragments thereof as described herein can also be assessed by measuring vδ1+ cell mediated modulation of other immune cells. For example, after application of an antibody or antigen binding fragment thereof as described herein to a model system comprising a mixed immune cell population (e.g., a mixed immune cell population comprising human tissue αβ cells and γδ T cells), the changes observed in the non- γδ T cell 'fraction' can be measured. Furthermore, the effect on the non- γδ cell types in the model can be measured by flow cytometry. For example, by measuring the relative change in the number of cd8+ αβ T cells after adding an antibody or antigen binding fragment thereof as described herein to a mixed culture comprising γδ T cells and non- γδ T cells. Optionally, the observed change in the number or phenotype of the non- γδ T cell cd8+ lymphocyte population can then be compared to the change in the number when an alternative comparison antibody (e.g., OKT-3) is applied to the mixed population.

Thus, in another aspect of the invention, there is provided a method of assessing an antibody or antigen binding fragment thereof that binds to a vδ1 chain of a γδ TCR, the method comprising administering the antibody or antigen binding fragment thereof to a mixed immune cell or tissue population comprising vδ1+ cells and vδ1 negative immune cells, and measuring the effect on the vδ1 negative immune cells. The effect may be determined/measured over a period of time and optionally compared to the effect observed in vδ1 negative cells over the same period of time without the antibody applied. The effect can be measured as a change in the number of vδ1 negative immune cells. For example, an antibody can increase the number of vδ1 negative immune cells by more than about 10%, more than about 20%, more than about 30%, more than about 40%, more than about 50%, more than about 70%, more than about 80%, more than about 90%, more than about 100%, more than about 500% relative to the level observed without the antibody.

In another aspect of the invention, the modulated vδ1 negative cells are cd45+ cells. In another aspect of the invention, the modulated cells are αβ T cells. In another aspect of the invention, the modulated αβ+ cells are cd8+ lymphocytes. In another aspect of the invention, the modulated αβ T cells or populations thereof exhibit evidence of enhanced cell division. In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by administering the antibody to a mixed immune cell population comprising vδ1+ cells and vδ1 negative immune cells, and then measuring the effect of the vδ1+ cells modulated by the antibody or antigen binding fragment thereof on the vδ1 negative cell population.

Optionally, and during "vδ1+ cell mediated modulation of the immune system" conferred by an antibody or antigen binding fragment thereof as described herein, a concomitant increase in vδ1+ cell number is also observed. While not being bound by this theory, it is possible that the increase in vδ1+ cell number may be responsible for the concomitant expansion of co-existing vδ1 negative immune cells (e.g. αβt cells). Another hypothesis might be that antibody-induced cytokine secretion from vδ1+ T cells stimulated expansion of vδ1 negative immune cells.

In another aspect of the invention, the observed increase in the αβ+cd8+ lymphocyte population is compared to a comparison antibody, such as an OKT3 antibody or alternatively an anti-vδ1 antibody. In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by applying the antibody to a mixed immune cell population comprising vδ1+ T cells and αβ T cells, and then measuring the number of cd8+αβ+ T cell lymphocytes over time.

Medicine for regulating Tumor Infiltrating Lymphocyte (TIL)

Antibodies or antigen binding fragments thereof as described herein can also be assessed by measuring the effect of conferring tumor infiltration populations (TILs) in a model system. Unexpectedly (see, e.g., example 18), antibodies as described herein measurably modulate the TIL population in human tumors during such an assessment. For example, after application of an antibody or antigen binding fragment thereof as described herein to a human tumor, such as human renal cell carcinoma, a change in the number or phenotype of a γδ+ lymphocyte TIL population or a non- γδ lymphocyte TIL population is measured. Optionally, the observed change in the number or phenotype of the γδ+ lymphocyte TIL population or the non- γδ lymphocyte TIL population can then be compared to the change observed when a surrogate comparison antibody (e.g., OKT-3) is applied to the model system.

Thus, in another aspect of the invention, there is provided a method of assessing antibodies or antigen binding fragments thereof that bind to the vδ1 chain of γδ TCRs, the method comprising administering the antibodies or antigen binding fragments thereof to a TIL located or derived from a human tumor and determining the effect on the number of TILs. The effect may be determined/measured over a period of time and optionally compared to the amount of TIL observed over the same period of time without the antibody applied. The effect may be an increase in the number of TILs. For example, an antibody may increase the amount of TIL by more than about 10%, more than about 20%, more than about 30%, more than about 40%, more than about 50%, more than about 70%, more than about 80%, more than about 90%, more than about 100% relative to the amount of TIL observed without the antibody. In another aspect, the observed number of TILs are γδ+ lymphocyte TIL cells and/or non- γδ lymphocyte TIL cells.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR cell antibody as described herein by applying the antibody to one or more TILs located or derived from a human tumor and then measuring the change in the number of one or more TILs over a period of time.

Medicament for regulating human V delta 1+ cytotoxicity

Antibodies or antigen binding fragments thereof as described herein can also be assessed by measuring the effect on vδ1+ mediated cytotoxicity conferring. Surprisingly, in such an evaluation procedure of antibodies as described herein (see, e.g., examples 19 and 27), measurable enhanced vδ1+ -mediated cytotoxicity was observed. For example, upon application of the antibody or antigen binding fragment thereof to a model system comprising a mixed culture comprising vδ1+ cells and the cancer cells, a decrease in the number of cancer cells or an increase in the number of cancer cells killed is observed. Optionally, a decrease in the number of cancer cells or an increase in the number of cancer cells killed can then be compared to the results when an alternative comparison antibody (e.g., OKT-3) is applied to the model system.

Thus, in another aspect of the invention, there is provided a method of assessing an antibody or antigen binding fragment thereof that binds to a vδ1 chain of a γδ TCR, the method comprising applying the antibody or antigen binding fragment thereof to a mixed population of cells comprising vδ1+ cells and cancer cells and measuring cytotoxicity of the vδ1+ cells to the cancer cells. Cytotoxicity may be measured by an increase in the number of dead cancer cells over a period of time, optionally compared to the number of dead cancer cells observed over the same period of time when the antibody is not applied to the mixed cell population. For example, the increase in dead cells observed with the antibody relative to the number of dead cells observed without the antibody can be greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 70%, greater than about 80%, greater than about 90%, greater than about 100%, greater than about 200%, greater than about 500%.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by adding the antibody to the mixed immune cell population comprising human vδ1+ cells and cancer cells, and then measuring the increase in dead cancer cells over time.

Agents that modulate the ratio of V delta 1+ cell target to effector cells (T: E ratio)

Antibodies or antigen binding fragments thereof as described herein can also be assessed by measuring how the antibodies enhance vδ1+ mediated cancer cell cytotoxicity by determining the ratio of target cells to effector cells, wherein 50% of target cells (EC 50) are killed in a model system, which can assess the antibodies as potential drugs. For example, a mixed culture comprising target cancer cells and human vδ1+ effector cells. Surprisingly, during such an evaluation procedure (see, e.g., example 19), antibodies as described herein advantageously alter the EC 50T: E ratio in the model system. Such modifications can be measured as the number of vδ1+ cells required to observe 50% cancer cell killing over a set period of time. This can also be reported as a fold or percentage improvement in cytotoxicity to the cancer cells. Optionally, the T:E ratio conferred by the antibodies of the invention can then be compared to the T:E ratio when an alternative comparison antibody (e.g., OKT-3) is applied to the model system. In some cases, the multispecific antibodies of the present invention provide an opportunity to improve cancer cell cytotoxicity as compared to monospecific antibodies, even at lower E: T ratios.

Thus, in another aspect of the invention, there is provided a method of assessing antibodies or antigen-binding fragments thereof that bind to the vδ1 chain of γδ TCR, the method comprising applying the antibodies or antigen-binding fragments thereof to a mixed population of cells comprising human vδ1+ cells and cancer cells and measuring the number of vδ1+ cells required to kill 50% of the cancer cells. This can be measured relative to the number of vδ1+ cells required to kill 50% of the cancer cells without the application of the antibody, optionally within the same time period. For example, the number of vδ1+ cells required to kill 50% of cancer cells when the antibody is applied may be reduced by more than about 10%, more than about 20%, more than about 30%, more than about 40%, more than about 50%, more than about 70%, more than about 80%, more than about 90%, more than about 100%, more than about 200%, more than about 500% relative to the number of vδ1+ cells required to kill 50% of cancer cells when the antibody is not applied.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to a vδ1 chain as described herein by adding the antibody to the population of cells comprising vδ1+ cells plus cancer cells, and then measuring the vδ1+ cells required to kill 50% of the cancer cells.

Medicament for enhancing cytotoxicity of V delta 1+ cell EC50

Another method of measuring the observed enhanced cytotoxicity of human vδ1+ cells or populations thereof is to measure the number of cells required to kill 50% of the cancer cells in a set period of time in condition a (e.g. the starting control) and compare this to the number of cells required to kill 50% of the cancer cells in a set period of time in condition B (e.g. when the antibodies of the present invention as described herein are applied).

While it is recognized that there are a variety of ways in which such parameters may be measured, to aid understanding, the following non-limiting hypothetical examples will be summarized:

It is hypothesized that the cytotoxicity enhancement of effector cells can be measured as follows: in condition a (control treatment), 1000 vδ1+ cells were observed to kill 50% of the cancer cells within a set period of time (e.g. 5 hours). In condition B (e.g., using the antibodies of the invention described herein), 500 vδ1+ cells were observed to be required to kill 50% of the cancer cells over the same period of time. Thus, in this example, the use of antibodies enhances cytotoxicity of the vδ1+ cell population by 200%:

(1000/500)x100＝200％

For example (see, e.g., examples 19 to 21), such a percent enhancement has been unexpectedly observed for antibodies of the invention as described herein.

In another aspect of the invention, there is provided a method of selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to the vδ1 chain of a γδ TCR as described herein by adding the antibody to the mixed immune cell population comprising vδ1+ cells and cancer cells and then determining the relative change or percentage of change in cytotoxicity relative to an equivalent or control experiment in which the antibody was not applied to the cell mixture.

Medicament for enhancing specificity of V delta 1+ cytopathic cells while preserving healthy cells

Another method of evaluating an antibody or antigen binding fragment thereof as described herein is to measure how the antibody modulates diseased cell-specific cytotoxicity. Surprisingly, during such studies, it was found that such antibodies can specifically enhance vδ1+ cell-specific killing of diseased cells such as cancer cells (see, e.g., examples 19 and 27) while retaining healthy or non-diseased cells. An ideal antibody drug to be administered to a patient to ameliorate symptoms of cancer would confer enhanced cytotoxicity specific to diseased cells while retaining healthy cells. And it can be said that a drug that enhances effector cell cytotoxicity specific to a diseased cell such as a cancer cell exhibits an enhanced Therapeutic Index (TI) than a drug that does not selectively enhance effector cell cytotoxicity specific to the diseased cell. The therapeutic index, also known as the therapeutic ratio, is a quantitative measure of the relative safety of a drug. It is the ratio of the amount of therapeutic agent that causes a therapeutic effect to the amount that causes toxicity (e.g., by causing undesired death in a related or associated healthy cell population). Antibodies or antigen binding fragments thereof as described herein can be evaluated by measuring their ability to alter or enhance or doubly improve the selective killing of diseased cells but not healthy cells in a vδ1+ cell selective killing model system. For example, the model system can comprise vδ1+ effector cells, cancer cells, and control cells (e.g., healthy cells). Optionally, the fold improvement in selective attenuation cell killing conferred by the antibodies of the invention can then be compared to the fold improvement observed when an alternative comparison antibody (e.g., OKT-3) is applied to the model system.

The diseased cell specificity and enhancement of diseased cell specificity of vδ1+ cells can be measured in cultures comprising vδ1+ cells, diseased cells, and healthy cells. For example, the specificity of vδ1+ for diseased cells can be measured by observing the number of cancer cells killed by vδ1+ cells and then comparing with the number of healthy cells killed by vδ1+ cells. Such comparison can be controlled by including equal amounts of diseased cells and healthy cells in a model system (e.g., "ternary cultures") that also contains vδ1+ cells. Alternative comparison methods are also contemplated, for example, when analysis or device limitations reduce the ability to distinguish and track all three or more cell types (including vδ1+ cells, diseased cells, and non-diseased cells) in parallel in a single assay. In that case, comparing the cytotoxicity of vδ1+ cells to diseased cells in one experiment, and then to healthy cells in another independent equivalent experiment, provides another approach to such studies.

In another aspect of the invention, there is provided a method of evaluating an antibody or antigen-binding fragment thereof that binds to a vδ1 chain of a γδ TCR, the method comprising administering the antibody or antigen-binding fragment thereof to a population of cells comprising vδ1+ cells and a target cell, and measuring cytotoxicity specificity for the target cell. In one embodiment, the cytotoxicity specificity for a first target cell type can be compared to the observed cytotoxicity for a second target cell type, and thus the method can be repeated using a different target cell type. In another aspect of the invention, the first target cell type is a diseased cell and the second target cell type is a control cell, e.g., a healthy cell or a cell having a different disease than the first target cell type.

In another aspect of the invention, there is provided a method for selecting or characterizing or comparing an antibody or antigen binding fragment thereof that binds to a vδ1 chain of a γδ TCR as described herein, wherein the effect on cytotoxicity of vδ1+ cells against (i) a first cell type and (ii) a second cell type conferred on the antibody is measured and compared. In another aspect of the invention, antibodies are thus selected that have a higher increase in specific cytotoxicity against a first cell type than against a second cell type. In another aspect of the invention, the first cell type is a diseased cell and the second cell type is a healthy cell.

As described herein, antibodies or antigen binding fragments thereof used in the assays may be presented on a surface, such as the surface of a cell (e.g., a cell comprising an Fc receptor). For example, the antibody or antigen binding fragment thereof may be presented on the surface of THP-1 cells, such as TIB-202 ^TM cells (available from American Type Culture Collection (ATCC)). Or the antibody or antigen binding fragment thereof may be used directly in the assay.

In such functional assays, output can be measured by calculating a half maximum concentration (also known as "EC50" or "effective concentration at 50%). The term "IC50" refers to inhibitory concentration. Both EC50 and IC50 may be measured using methods known in the art, such as flow cytometry methods. In some cases, the EC50 and IC50 are the same value or may be considered equivalent. For example, the Effective Concentration (EC) of effector cells required to inhibit (e.g., kill) 50% of a cell type can also be considered as a 50% Inhibition Concentration (IC). For the avoidance of doubt, when referring to antibodies, the EC50 values in the present application are provided using antibodies in the IgG1 format. Such values can be readily converted to equivalent values based on the molecular weight of the antibody format as follows:

(μg/ml)/(MW (in kDa)) =μM

For the parental clones described herein, the EC50 for γδ TCR downregulation after antibody (or fragment) binding may be less than 0.50 μg/ml, e.g., less than 0.40 μg/ml, 0.30 μg/ml, 0.20 μg/ml, 0.15 μg/ml, 0.10 μg/ml, 0.06 μg/ml, or 0.05 μg/ml. In a preferred embodiment, the γδ TCR is down-regulated with an EC50 of less than 0.10 μg/ml after antibody (or fragment) binding. In particular, the EC50 for downregulating γδ TCR after antibody (or fragment) binding may be less than 0.06 μg/ml, e.g., less than 0.05 μg/ml, 0.04 μg/ml, or 0.03 μg/ml. In particular, the EC50 value is the value when the antibody is measured in IgG1 format. For example, γδ TCRs can be measured using flow cytometry to down-regulate EC50 values (e.g., as described in the assays of examples 6 and 27, for example).

The EC50 for degranulation of γδ T cells after antibody (or fragment) binding may be less than 0.050 μg/ml, e.g., less than 0.040 μg/ml, 0.030 μg/ml, 0.020 μg/m1, 0.015 μg/ml, 0.010 μg/ml, or 0.008 μg/ml. In particular, the EC50 for degranulation of γδ T cells after antibody (or fragment) binding can be less than 0.005 μg/ml, e.g., less than 0.002 μg/ml. In a preferred embodiment, the EC50 of degranulation of γδ T cells after antibody (or fragment) binding is less than 0.007 μg/ml. In particular, the EC50 value is the value when the antibody is measured in IgG1 format. For example, γδ T cell degranulation EC50 values can be measured by detecting CD107a expression (i.e., a marker of cell degranulation) using flow cytometry (e.g., as described in the assay of example 7). In one embodiment, CD107a expression is measured using an anti-CD 107a antibody, e.g., anti-human CD107a BV421 (clone H4 A3) (BD Biosciences).

The EC50 of γδ T cell killing after antibody (or fragment) binding may be less than 0.50 μg/ml, e.g. less than 0.40 μg/ml, 0.30 μg/ml, 0.20 μg/ml, 0.15 μg/ml, 0.10 μg/ml or 0.07 μg/ml. In a preferred embodiment, the EC50 of γδ T cell killing after antibody (or fragment) binding is less than 0.10 μg/ml. In particular, the EC50 of γδ T cell killing after antibody (or fragment) binding may be less than 0.060 μg/ml, such as less than 0.055 μg/ml, especially less than 0.020 μg/ml. In particular, the EC50 value is the value when the antibody is measured in IgG1 format. For example, γδ T cell killing EC50 values can be measured by detecting the proportion of dead cells (i.e., using cell viability dye) using flow cytometry after incubation of the antibody, γδ T cells, and target cells (e.g., as described in the assay of example 8). In one embodiment, the death of the target cells is measured using a cell viability dye, namely the viability dye eFluor ^TM (ThermoFisher).

In the assays described in these aspects, the antibodies or antigen binding fragments thereof may be presented on the surface of cells, such as THP-1 cells (e.g., TIB-202 ^TM (ATCC)). THP-1 cells are optionally labeled with a dye, such as CellTracker ^TM Orange CMTMR (thermo fisher).

Agents that down-regulate CD3 molecules associated with vδ1 TCR

Antibodies that bind T cell/CD 3 complexes in different ways are presented herein. In particular, these antibodies can be conjugated via the TRDV1 domain of vδ1 TCR expressed on vδ1+ cells only. In so doing, such drugs act in different ways. This engagement event in turn down-regulates the TCR-associated CD3 molecule complex. Such CD3 down-regulation may be synonymous with T cell activation. However, by engaging the T-cell/CD 3 complex via the TRDV1 domain in this manner, only CDR3 molecules associated with vδ TCR are down-regulated. This mechanism is clearly shown in fig. 55.

Thus, in one embodiment, a method of down-regulating a vδ1 TCR and related CD3 molecular complex containing TRDV1 on the cell surface with an antibody is provided, as well as uses of such antibodies for this purpose.

In some embodiments, also presented herein are multi-specific TCEs capable of engaging T cell/CD 3 complexes via TRDV domain. Current drugs in the multispecific TCE format typically bind and activate T cells via a CD3 binding event. This can result in down-regulation of the CD3 molecular complex from the T cell surface. However, it is also clear that TCE can also overstimulate T cells via such engagement and down-regulation. CD3 molecular complexes are not specific for a class of T cells and are therefore not a precise target. Stimulation of all T cells via CD3 (predominantly the αβ subtype) can in turn lead to cytokine overproduction, leading to acute cytokine burst (so-called cytokine storm). Furthermore, in non-targeted approaches to engaging and activating all T cells via CD3, it is paradoxical that T cells may be overactivated, resulting in chronic T cell depletion and/or T cell death. In fact, this non-specific T cell activation results in activation of effector T cells and regulatory T cells, whereas the presently proposed method specifically interacts with effector populations. Thus, this 'sledge' approach is far from ideal when one might wish to up-regulate only selective T cells.

In contrast, multi-specific antibodies, particularly T cell cements, are presented herein that bind T cell/CD 3 complexes in a different manner. In particular, these TCEs can be joined via the TRDV1 domain of vδ1 TCR expressed on vδ1+ cells only. In so doing, such TCE-based drugs function in different ways. First, these TCEs are able to down-regulate the TCR via the binding TRDV-1 epitope. This engagement event in turn down-regulates the TCR-associated CD3 molecule complex. Such CD3 down-regulation may be synonymous with T cell activation. However, by engaging the T-cell/CD 3 complex via the TRDV1 domain in this manner, only CDR3 molecules associated with vδ TCR are down-regulated. This approach of specifically targeting and activating vδ1 cells can avoid many of the problems described above (e.g. cytokine storm, T cell depletion/depletion and ADCC). This mechanism is again clearly shown in fig. 55.

Stimulation of T cells via the CD3 'sledgehammer' method may also promote T cell depletion via fcγ receptor driving mechanisms (e.g., ADCC). Thus, most CD 3-targeted bispecific antibodies in current clinical practice have an Fc domain with reduced binding activity to fcγr, or are bispecific fragments that intentionally lack an Fc region. CD3 targeted therapies may also have reduced binding affinity of the T cell receptor complex binding arms.

This decrease in affinity can lead to reduced efficacy and reduced selectivity in TCE design and function. For example, the affinity of binding domains in such TCEs is known to drive in vivo distribution properties. In particular, TCE distribution is often observed biased towards its highest affinity target. Thus, by decreasing the affinity of the TCE binding domain to the T cell complex, a distribution away from T cells is generally favored; these T cells are the cells required to drive the efficacy of such TCEs. Partly for these reasons, the TCE therapeutic window is referred to as 'too narrow'.

The methods presented herein specifically target and activate vδ1 cells, avoiding the need to eliminate Fc function or reduce affinity.

Furthermore, in the methods presented herein, the antibody binds to the TCR of the vδ1 cell, but no complete activation occurs unless tumor cells are also present. Complete engagement of the TCR by the currently presented antibodies results in partial downregulation, and the v delta 1 cells bound by the currently presented antibodies are fully activated and cytotoxic only in the tumor microenvironment. This represents another important safety advantage for the methods of the invention, as off-target cytotoxicity is reduced and the full efficacy of the antibodies of the invention to activate vδ1 cells is only released in the presence of tumor cells.

One mechanism behind γδ T cells being able to detect stress signals on tumor cells is thought to be due to the NCR (natural cytotoxic receptor) they express. NCR is capable of binding NCR ligands on tumor cells. Thus a dual activation mechanism is employed, where γδ T cells are activated via TCR stimulation, and NCR perceives tumor cells to allow complete activation and cytotoxicity.

This is in contrast to stimulation of αβ T cells via CD3, where all stimulation is via TCR. Thus, such cells hardly distinguish between healthy cells or transformed cells, as they do not have this antigen presentation independent perception of tumor cells via NCR. Thus, if CD3 antibodies are Fc-effective, they will attract other immune cells, which may trigger a series of unpredictable and desirable events, such as cytokine storms, immune cell depletion and even overactivation, leading to, for example, NK cell killing T cells, etc. In the methods of the invention, stimulation of γδ T cells with the presently presented antibodies does not raise such concerns, as γδ T cells (and other immune cells, such as NK cells) are able to distinguish healthy cells from tumor cells, including via their NCR sensing mechanism, and thus selectively kill stressed cells, such as cancer cells or virally infected cells, due to the specificity of such diseased cells.

For example, in an initial cynomolgus study with a multispecific antibody effective against both human and cynomolgus vδ1 antigen presented herein (e.g. SEQ ID NO:272 and SEQ D NO: 308) that is specific for vδ1 Fc, it was found to be safe and well tolerated in vivo studies of repeated doses of ascending doses, based on all measured parameters. No side effects normally associated with T cell activation, such as cytokine release or weight loss, were observed.

These findings also highlight another advantage of the methods described herein. In particular, unlike TCEs characterized by CD3 binding agents and the like, TCE bispecific antibodies of the present invention can optionally be designed as full length antibodies, e.g., comprising a heavy chain having VH-CH1-CH2-CH3 format and a cognate light chain having VL-CL format. Unlike smaller bispecific formats (e.g., less than 70 KDa), such full-length bispecific formats can exhibit longer in vivo half-lives, thus requiring less frequent dosing regimens. The longer half-life observed in such formats has several reasons, including increased size (> 70 kDa), which means that such formats are not filtered by the kidneys (glomerular pore size cut-off 60-70 kDa). In one embodiment, the multispecific antibody may be greater than about 70KDa, and may comprise a human IGHC sequence (e.g., IGHA, IGHD, IGHD, IGHM, IGHG sequence) as listed in IMGT. Such IgG1 formats may also be recycled through the FcRn mechanism. The clear disadvantage of the full length antibody format of such TCE bispecific antibodies is in particular that such formats exhibit disadvantageous safety profiles due to reduced clearance, increased exposure and longer time.

Thus, the method of the invention allows the possibility of Fc function without concern for off-target effects, such as NK cell killing γδ T cells, and vice versa. Thus, the methods of the invention are superior to CD3 targeting methods, which are hindered by the need for alternative methods, such as reduced CD3 affinity, elimination of Fc function, etc., to limit collateral damage outside of the tumor environment. The multispecific antibodies presented herein, particularly T cell cements, are capable of binding to vδ1 cells without any potential damage, and will only fully activate and have enhanced cytotoxicity when vδ1 cells are in intimate contact with tumor cells.

Thus, in one embodiment, a method is provided for down-regulating a vδ1 TCR and related CD3 molecular complex containing TRDV1 on the cell surface with a TCE multispecific antibody, and the use of such a multispecific antibody for achieving this purpose.

Other features and advantages of the present invention will be apparent from the description provided herein. It should be understood, however, that the description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications will become apparent to those skilled in the art. The invention will now be described using the following non-limiting examples:

Examples

Example 1 materials and methods

Human antibody discovery

Human phage display was used to generate human anti-human variable vδ1+ domain antibodies as described herein. The library was constructed as described in Schofield et al (Genome biology 2007,8 (11): R254) and contained about 400 million human cloned single chain fragment variable (scFv) display libraries. The library was screened using the antigen, method, selection, deselection, screening and characterization strategies as described herein.

Antigen preparation

The soluble ydelta TCR heterodimers comprising TCR α and TCR β constant regions used in the following examples were designed according to Xu et al (2011) PNAS108: 2414-2419. The vγ or vδ domain is fused in frame with a TCR α or TCR β constant region lacking the transmembrane domain, followed by a leucine zipper sequence or Fc sequence and a histidine tag/linker.

The expression constructs were transiently transfected into mammalian EXPI HEK293 suspension cells (as single transfection or co-transfection for heterodimers). Secreted recombinant proteins were recovered from the culture supernatant by affinity chromatography and purified. To ensure good recovery of monomeric antigen, the samples were further purified using preparative Size Exclusion Chromatography (SEC). Purified antigen was analyzed for purity by SDS-PAGE and aggregation status by analytical SEC.

Antigen functional validation

The specificity of antigen containing delta variable 1 (vδ1) chains was demonstrated in competition with γδ T cells in DELFIA immunoassay (PERKIN ELMER) and flow cytometry-based assays using the REA173-Miltenyi Biotec anti-vδ1 antibodies.

Dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA)

To confirm antigen specificity, DELFIA immunoassays were performed by directly coating antigen onto plates (3. Mu.g/mL antigen in 50. Mu.L PBS overnight at 4 ℃ (Nunc# 437111) and serial dilution of primary antibody starting at 300nM for detection DELFIA Eu-N1 anti-human IgG (PERKIN ELMER #1244-330) was used as secondary antibody, diluted 1/500 in 50. Mu.L 3% MPBS (PBS+3% (w/V) skimmed milk powder) and developed using 50. Mu.L DELFIA enhancement solution (PERKIN ELMER #4001-0010).

Affinity sequencing was performed on the antibodies of interest using a DELFIA immunoassay, in which the antibodies were captured via protein G coated on a plate, and 5nM of soluble biotinylated L1 (DV 1-GV 4) antigen in 50 μl (3 MPBS) was added. For detection, 50 μl of streptavidin-Eu (1:500,Perkin Elmer in assay buffer) was used and the signal was developed with DELFIA enhanced solution. D1.3hLgG1 (described in England et al (1999) J.Immunol.162:2129-2136) was used as a negative control.

Phage display selection output was subcloned into scFv expression vector pSANG (Martin et al (2006) BMC Biotechnol.6:46). Soluble scFv were expressed and screened for binding to directly immobilized targets in DELFIA. Hits were defined as DELFIA signals above 3000 fluorescent units.

Antibody preparation

Selected scFv were subcloned into IgG1 frames using commercially available plasmids. The plasmid was used to transfect expi293F suspension cells for antibody expression. For convenience, the antibodies expressed in these examples refer to antibodies in the IgG1 format selected as scFv from phage display, unless otherwise specified. However, the antibodies of the invention may be in any antibody format as previously discussed.

Antibody purification

IgG antibodies were purified in bulk from the supernatant using protein a chromatography. The concentrated protein a eluate is then purified using Size Exclusion Chromatography (SEC). The quality of purified IgG was analyzed using ELISA, SDS-PAGE and SEC-HPLC.

Gamma delta T cell preparation

Enriched γδ T cell populations were prepared according to the methods described in WO2016/198480 (i.e., blood-derived γδ T cells) or WO2020/095059 (i.e., skin-derived γδ T cells). Briefly, for blood-derived γδ T cells, PBMCs are obtained from blood and αβ T cells are magnetically depleted. The alpha beta depleted PBMC were then incubated in CTS OpTmiser medium (ThermoFisher) in the presence of OKT-3 (or respective anti-V delta 1 antibodies), IL-4, IFN-gamma, IL-21 and IL-1β for 7 days. On day 7 of culture, the medium was supplemented with OKT-3 (or the respective anti-V.delta.1 antibodies), IL-21 and IL-15 for an additional 4 days. On day 11 of culture, the medium was supplemented with OKT-3 (or the respective anti-V.delta.1 antibody) and IL-15 for an additional 3 days. On day 14 of culture, half of the medium was replaced with fresh complete OpTmiser and supplemented with OKT-3 (or respective anti-V.delta.1 antibodies), IL-15 and IFN-gamma. Cultures were supplemented with OKT-3 (or respective anti-V.delta.1 antibodies) and IL-15 every 3 to 4 days, starting on day 17 of culture; half of the medium was replaced with fresh medium every 7 days.

For skin-derived γδ T cells, skin samples were prepared by removing subcutaneous fat and perforated multiple times using a 3mm biopsy punch. The perforator was placed on a carbon matrix grid and in the holes of G-REX6 (Wilson Wolf). Each well was filled with complete isolation medium containing AIM-V medium (Gibco, life Technologies), CTS immune serum replacement (Life Technologies), IL-2 and IL-15. At the first 7 days of culture, a complete isolation medium containing amphotericin B (Life Technologies) ("+amp") was used. The medium was changed every 7 days by gently sucking out the upper medium and replacing it with 2X complete isolation medium (without AMP) with minimal disruption to the cells on the plate or bottom of the bioreactor. After more than three weeks of culture, the resulting drained cells were passaged into fresh tissue culture vessels and fresh medium (e.g., AIM-V medium or TexMAX medium (Miltenyi)) plus recombinant IL-2, IL-4, IL-15 and IL-21 prior to harvest. Optionally, the αβ T cells also present in the culture are then removed by means of an αβ T cell depletion kit and related protocols (e.g., protocols provided by Miltenyi). For further reference, see WO2020/095059.

Gamma delta T cell binding assay

Antibody binding to γδ T cells was tested by incubating purified antibody at a fixed concentration with 250000 γδ T cells. This incubation is performed under blocking conditions to prevent non-specific binding of the antibody via the Fc receptor. Detection was performed by adding a fluorescent dye conjugated secondary antibody to human IgG 1. For negative controls, cells were prepared as follows: a) isotype antibody only (recombinant human IgG), b) fluorescent dye conjugated anti-human IgG antibody only, and c) a) in combination with b). Control wells of completely unstained cells were also prepared and analyzed. As positive controls, two different concentrations of purified murine monoclonal IgG2 anti-human CD3 antibody and purified murine monoclonal IgG1 anti-human TCR vδ1 antibody were used and stained with a fluorochrome conjugated goat anti-mouse secondary antibody. The assay is accepted if the average fluorescence intensity of the lower concentration positive control in the FITC channel is at least 10 times that of the highest negative control.

SPR analysis

MASS-2 instrument (all from Sierra Sensors, germany) with amine high capacity chip was used to perform SPR analysis. 15nM IgG was captured via protein G to an amine high capacity chip (100 nM for TS 8.2). L1 (DV 1-GV 4) antigen was passed through cells in a 1:2 dilution series of 2000nM to 15.625nM with the following parameters: 180 seconds association, 600 seconds dissociation, flow rate 30. Mu.L/min, running buffer PBS+0.02% Tween 20. All experiments were performed on a MASS-2 instrument at room temperature. Steady state fits were determined from langmuir 1:1 combinations using the software Sierra Analyzer 3.2.

Comparison antibody

The antibodies of the invention were compared to commercially available antibodies in a test assay as described.

Gamma delta TCR down regulation and threshing assay

THP-1 (TIB-202 ^TM, ATCC) target cells, with or without test antibody, were labeled with CellTracker ^TM orange CMTMR (ThermoFisher, C2927) and incubated with γδ T cells at 2:1 in the presence of CD107a antibody (anti-human CDl07a BV421 (clone H4 A3) BD Biosciences 562623). After 2 hours of incubation, surface expression of γδ TCR on γδ T cells (to measure TCR down) and expression of CD107a (to measure degranulation) were assessed using flow cytometry.

Killing assay (e.g. FIG. 6)

The effect of the test antibodies on γδ T cell killing activity was assessed by flow cytometry. After 4 hours of in vitro co-culture, γδ T cells and CellTracker ^TM orange CMTMR (ThermoFisher, C2927) labeled THP-1 cells (loaded or unloaded) were stained with the vital dye eFluor ^TM 520 (ThermoFisher, 52065-0867-14) in a 20:1 ratio to distinguish between live and dead target THP-1 cells. During sample collection, target cells were gated according to CellTracker ^TM orange CMTMR positives and examined for cell death based on uptake of vital dyes. Cmmr and eFluor ^TM 520 biscationic cells were identified as dead target cells. The killing activity of γδ T cells is presented as% of dead target cells.

Epitope mapping

All protein samples (antigen L1 (DV 1-GV 4) and antibodies 1245_p01_e07, 1245_p02_g04, 1252_p01_c08, 1251_p02_c05 and 1141_p01_e01) for epitope localization were analyzed for protein integrity and aggregation levels using high quality MALDI.

To determine the epitope of the L1 (DV 1-GV 4)/1245_p01_e07, L1 (DV 1-GV 4)/1245_p02_g04, L1 (DV 1-GV 4)/1252_p01_c08, L1 (DV 1-GV 4)/1251_p02_c05 and L1 (DV 1-GV 4)/1141_p01_e01 complex at high resolution, the protein complex is incubated with deuterated cross-linking agent and subjected to multi-enzyme proteolysis using trypsin, chymotrypsin, asp-N, elastase and thermolysin. After enrichment of the cross-linked peptides, the samples were analyzed by high resolution mass spectrometry (nLC-LTQ-Orbitrap MS) and the resulting data was analyzed using XQuest and Stavrox software.

SYTOX-flow killing assay

The SYTOX assay allows for the quantification of T cell mediated cytolysis of target cells using flow cytometry. Through dead cell stainAADVANCED ^TM, life Technologies, S10274) to detect dead/dying cells, the dye can only penetrate into cells with damaged plasma membranes and cannot penetrate the whole cell membrane of healthy cells. NALM-6 target cells were labeled with CTV dye (CELL TRACE Violet ^TM, life Technologies, C34557) and therefore distinguished from unlabeled effector T cells. Dead/dying target cells are identified by double staining of dead cell dyes and cell tracking dyes.

After 16 hours of co-culturing effector cells and CTV-labeled target cells in vitro at the indicated effector cell to target cell ratio (E: T, 1:1 or 10: 1), the cells were usedAADVANCED ^TM were stained and collected on FACSLYRIC ^TM (BD). The killing results were presented as target cell reduction, which was calculated by considering the number of live target cells in the test sample (sample count) relative to the live target cells in the control wells without effector cells added (maximum count):

target decrease% = 100- ((sample count/max count) x100

Example 2 antigen design

Γδ (γδ) T cells are polyclonal cells with CDR3 polyclonality. To avoid the situation where the generated antibody is selected for CDR3 sequences (as CDR3 sequences will differ between TCR clones), antigen design involves maintaining consistent CDR3 in different formats. This design was intended to generate antibodies that recognized variable domain sequences encoded by the germline and thus identical in all clones, providing antibodies that recognized a broader subset of γδ T cells.

Another important aspect of the antigen preparation process is the design of antigens suitable for expression as proteins. γδ TCRs are complex proteins involving heterodimers with inter-and intra-chain disulfide bonds. The soluble TCR antigen to be used for phage display selection was generated using Leucine Zipper (LZ) format and Fc format. Both LZ and Fc formats expressed well and successfully displayed TCRs (particularly heterodimeric TCRs, e.g., vδ1vγ4).

CDR3 sequences from the public database entry of γδ TCRs were found to perform as well as proteins (RCSB protein database entry: 3 OMZ). It was therefore selected for antigen preparation.

Antigens containing delta variable 1 chains are expressed as heterodimers in LZ format (i.e., in combination with a different gamma variable chain- "L1", "L2", "L3") and as heterodimers ("F1", "F2", "F3") or homodimers in Fc format (i.e., in combination with another delta variable 1 chain- "Fc 1/1"). All delta variable 1 chains of the antigen contained 3omz cdrs 3. Another series of γδ TCR antigens using a similar format were designed to contain different δ variable chains (e.g., δ variable 2 and δ variable 3) and were used to deselect antibodies ("L4", "F9", "Fc4/4", "Fc 8/8") that have non-specific or off-target binding. These antigens were also designed to include 3omz CDR3 to ensure that antibodies that bind in the CDR3 region were also deselected.

Antigen functional validation was performed to confirm that the designed antigen was suitable for the generation of anti TRDV1 (TCR delta variable 1) antibodies. Only antigen containing the δ1 domain was detected (fig. 1).

EXAMPLE 3 phage display

Phage display selection was performed on a human scFv library using either heterodimeric LZ TCR format in rounds 1 and 2, with heterodimeric LZ TCR deselection in both rounds. Or round 1 was performed using homodimeric Fc fusion TCRs, deselecting human IgG1 Fc, then round 2 was performed on heterodimeric LZ TCRs, deselecting heterodimeric LZ TCRs (see table 7).

TABLE 7 overview phage display selection

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Bt = biotin.

Selection was performed in the solution phase using 100nM biotinylated protein. Deselection was performed using 1 μm non-biotinylated protein.

The success of phage display selection was analyzed by polyclonal phage ELISA (DELFIA). All DV1 select outputs show the desired binding to targets Fc 1/1, L2, L3, F1 and F3. Different degrees of binding to non-targets L4, F9, fc 4/4, fc 8/8 and Fc were detected (see FIGS. 2A and B).

Example 4 antibody selection

The hits obtained in example 3 were sequenced (using standard methods known in the art). 130 unique clones were identified that showed a unique combination of VH and VL CDR3. Of these 130 unique clones, 125 showed unique VH CDR3 and 109 showed unique VL CDR3.

Unique clones were rearranged and analyzed for specificity by ELISA (DELFIA). From the selection, a panel of 94 unique human scFv conjugates that bound TRDV1 (L1, L2, L3, F1, F2, F3) but did not bind TRDV (L4) was identified.

Affinity ordering of the selected binders was included to aid in the forward progression of the selection of clones. A large number of conjugates showed affinities in the nanomolar range, reacting with 25 to 100nM biotinylated antigen. A small amount of conjugate showed a strong reaction with 5nM antigen, indicating that nanomolar affinities for unit numbers may exist. Some binders did not show reaction with 100nM antigen, indicating affinities in the micromolar range.

In order to select clones that underwent conversion to IgG, the objective was to incorporate as many germline lines and as many different CDR3 as possible. Further, sequence propensity such as glycosylation, integrin binding site, CD11c/CD18 binding site, unpaired cysteines are avoided. In addition, various affinities are also included.

Selected clones were screened for binding to native cell surface expressed γδ TCR using skin derived γδ T cells obtained from different donors. The clones selected to be converted to IgG are shown in table 8.

Example 5: antibody SPR analysis

IgG antibodies were prepared by γδ cell binding assay and 5 were selected for further functional and biophysical characterization. SPR analysis was performed to determine equilibrium dissociation constants (K _D). The sensorgram of the test antibody interacting with the analyte, if available, and the steady state fit are presented in figure 3. No binding of TS8.2 to on-chip captured 80 RU IgG was detected. The results are summarized in table 9.

TABLE 9 IgG capture results

* The binding of 1252_p02_c05 did not reach saturation and the data was extrapolated.

Example 6: TCR engagement assay

The inventors devised several assays for functional characterization of selected antibodies. The first assay evaluates γδ TCR engagement by measuring the down-regulation of γδ TCR after antibody binding. The selected antibodies were tested against commercial anti-CD 3 and anti-vδ1 antibodies used as positive controls or against 1252_p01_c08 (for 1139_p01_e04, 1245_p02_f07, 1245_p01_g06 and 1245_p01_g09) used as positive controls. Commercial anti-pan γδ was used as a negative control, since it is a pan γδ antibody, recognizing all γδ T cells irrespective of the variable chain, and thus may have different modes of action.

Assays were performed using skin-derived γδ T cells obtained from three different donor samples (samples of 94%, 80% and 57% purity). The results are shown in fig. 4. EC50 values are summarized in table 10 below.

Example 7: t cell degranulation assay

The second assay evaluates degranulation of γδ T cells. Gamma delta T cells are thought to mediate target cell killing through perforin-granzyme mediated activation of apoptosis. Lysed particles within the gamma delta T cell cytoplasm can be released to target cells upon T cell activation. Thus, labelling target cells with antibodies to CD107a and measuring expression by flow cytometry can be used to identify degranulated γδ T cells.

For example 6, the selected antibodies were tested against commercial anti-CD 3 and anti-vδ1 antibodies as positive controls or against 1252_p01_c08 (for 1139_p01_e04, 1245_p02_f07, 1245_p01_g06, and 1245_p01_g09) as positive controls. IgG2a, igG1 and D1.3 antibodies were used as negative controls. Assays were performed using skin-derived γδ T cells obtained from three different donor samples (samples of 94%, 80% and 57% purity). The results are shown in fig. 5. EC50 values are summarized in table 10 below.

Example 8: killing assay

A third assay evaluates the ability of γδ T cells activated with the selected antibody to kill target cells.

For example 6, the selected antibodies were tested against commercial anti-CD 3 and anti-vδ1 antibodies as positive controls or against 1252_p01_c08 (for 1139_p01_e04, 1245_p02_f07, 1245_p01_g06 and 1245_p01_g09) as positive controls and anti-pan γδ as negative controls. IgG2a, igG1 and D1.3 antibodies were also used as isotype controls. Assays were performed using skin-derived γδ T cells obtained from two donors (94% and 80% purity), the results are shown in fig. 6.

The results of the three functional assays tested in examples 6-8 are summarized in table 5.

TABLE 10 summary of results obtained from functional assays

N/D: failing to determine; N/D: failing to determine that the titration curve does not reach plateau; N/D: reduced killing characteristics, EC50 was not established

Example 9: epitope mapping

To determine the epitope of the antigen/antibody complex at high resolution, the protein complex is incubated with deuterated cross-linking agents and subjected to multienzyme cleavage. After enrichment of the cross-linked peptides, the samples were analyzed by high resolution mass spectrometry (nLC-LTQ-Orbitrap MS) and the resulting data was analyzed using XQuest (version 2.0) and Stavrox (version 3.6) software.

After proteolytic cleavage of protein complex L1 (DV 1-GV 4)/1245_P01_E07 with deuterated d0d12 by trypsin, chymotrypsin, asp-N, elastase and thermolysin, nLC-orbitrap MS/MS analysis detected 13 cross-linked peptides between L1 (DV 1-GV 4) and antibody 1245_P01_E07.

After proteolytic cleavage of protein complex L1 (DV 1-GV 4)/1252_P01_C08 with deuterated d0d12 by trypsin, chymotrypsin, asp-N, elastase and thermolysin, nLC-orbitrap MS/MS analysis detected 5 cross-linked peptides between L1 (DV 1-GV 4) and antibody 1252_P01_C08.

After proteolytic cleavage of protein complex L1 (DV 1-GV 4)/1245_P02_G04 with deuterated d0d12 by trypsin, chymotrypsin, asp-N, elastase and thermolysin, nLC-orbitrap MS/MS analysis detected 20 cross-linked peptides between L1 (DV 1-GV 4) and antibody 1245_P02_G04.

After proteolytic cleavage of the protein complex L1 (DV 1-GV 4)/1251_P02_C05 with deuterated d0d12 by trypsin, chymotrypsin, asp-N, elastase and thermolysin, nLC-orbitrap MS/MS analysis detected 5 cross-linked peptides between L1 (DV 1-GV 4) and antibody 1251_P02_C05.

Binding to an epitope of another antibody clone ID 1141_p01_e01 was also tested. After proteolytic cleavage of protein complex L1 (DV 1-GV 4)/1141_P01_E01 with deuterated d0d12 by trypsin, chymotrypsin, asp-N, elastase and thermolysin, nLC-orbitrap MS/MS analysis detected 20 cross-linked peptides between L1 (DV 1-GV 4) and antibody 1141_P01_E01.

A summary of epitope mapping results is presented in table 11.

TABLE 11 epitope mapping results for antigen/antibody complexes

Clone ID	Epitope mapping, SEQ ID NO:272 amino acid numbering
		1245_P01_E07	5、9、16、20、62、64、72、77
1252_P01_C08	50、53、59、62、64
		1245_P02_G04	37、42、50、53、59、64、68、69、72、73、77
1251_P02_C05	59、60、68、72
		1141_P01_E01	3、5、9、10、12、16、17、62、64、68、69

Example 10: expansion of V.delta.1T cells

The expansion of isolated γδ T cells was studied in the presence of selected antibodies and comparison antibodies. The comparison antibody is selected from: OKT3 anti-CD 3 antibody was used as positive control, no antibody as negative control or IgG1 antibody as isotype control. Commercially available anti-V.delta.1 antibodies TS-1 and TS8.2 were also tested for comparison.

Experiment 1:

Initial studies were performed by seeding 70,000 cells/well with a complete optimizer and cytokines as described in example 1 in "γδ T cell preparation" for blood-derived γδ T cells. The selected antibodies and the comparison antibodies were tested at various concentrations ranging from 4.2ng/ml to 420 ng/ml. This experiment was performed using tissue culture plates, which allow antibodies to bind/immobilize to plastic.

Cells were harvested on days 7, 14 and 18 and total cell counts were determined using a cell counter (NC 250, chemoMetec). The results are shown in fig. 7. Cell viability of vδ l T cells was also measured at each harvest and all antibodies were shown to maintain cell viability throughout the experiment (data not shown). The percentage, cell count and fold change of vδ1T cells were also analyzed on day 18. The results are shown in fig. 8.

As can be seen in fig. 7, the total number of cells produced in the cultures with the antibodies steadily increased throughout the culture, comparable to or better than the commercial anti-vδ1 antibodies. On day 18, the proportion of vδ1 positive cells was higher in the presence of the majority of the test concentrations of the 1245_p02_g04 ("G04"), 1245_p01_e07 ("E07"), 1245_p01_b07 ("B07") and 1252_p01_c08 ("C08") antibodies than in cultures in the presence of OKT3, TS-1 or TS8.2 control antibodies (see fig. 8A).

Experiment 2:

Subsequent experiments were performed on isolated cells in culture vessels with cytokines as described in "γδ T cell preparation" of example 1. In comparison to experiment 1, a different culture vessel was used, the surface of which was unfavorable for antibody binding/immobilization. The selected antibodies and the comparison antibodies were tested at different concentrations ranging from 42pg/ml to 42 ng/ml. During experiment 2, results were obtained from triplicate experimental runs.

Cells were harvested on day 7, day 11, day 14 and day 17 and total cell counts were determined using a cell counter as described previously. The results are shown in fig. 9. The percentage, cell count and fold change of vδ1T cells were also analyzed on day 17. The results are shown in fig. 10.

Cell composition, including non-vδ1 cells, was also measured during experiment 2. Cells on day 17 were harvested and analyzed for surface expression of vδ1, vδ2 and αβtcr by flow cytometry. The proportion of each cell type in each culture is graphically shown in fig. 11, and the percentage values are provided in table 12.

TABLE 12 composition of cells on day 17-percentage of viable cells per subset

	αβ-γδ-	Vδ1	Vδ2	Non-V.delta.1/V.delta.2	αβ
						AB-free	63.00	18.17	0.86	7.10	0.37
OKT-3	25.63	50.43	0.25	20.13	1.13
						IgG1	65.77	15.59	1.11	6.91	0.42
TS8.2 42ng/ml	30.60	53.57	3.59	7.46	0.14
						TS-1 42ng/ml	18.77	65.90	0.91	9.51	0.12
C08 42ng/ml	0.79	96.43	0.08	2.51	0.05
						C08 4.2ng/ml	1.91	94.67	0.18	2.63	0.05
C08 420pg/ml	8.47	80.57	0.28	8.42	0.04
						C08 42pg/ml	35.97	25.93	3.04	19.50	0.31
B07 42ng/ml	0.94	95.57	0.46	2.73	0.05
						B07 4.2ng/ml	1.79	94.10	0.40	3.28	0.01
B07 420pg/ml	3.08	91.80	0.29	3.94	0.02
						B07 42pg/ml	17.93	62.90	0.85	9.16	0.07
E07 42ng/ml	2.29	85.13	0.19	11.65	0.04
						E07 4.2ng/ml	2.15	91.23	0.13	5.77	0.04
E07 420pg/ml	9.25	73.90	0.42	13.05	0.02
						E07 42pg/ml	49.23	18.67	2.17	7.70	0.43
G04 42ng/ml	1.90	88.53	0.47	8.09	0.05
						G04 4.2ng/ml	4.25	89.67	0.93	3.98	0.02
G04 420pg/ml	25.97	50.60	1.45	12.72	0.11
						G04 42pg/ml	44.00	13.77	2.33	26.30	0.32
C05 42ng/m1	25.00	42.03	3.75	13.67	1.32
						C05 4.2ng/ml	46.87	22.03	2.58	16.46	0.38
C05 420pg/ml	33.53	44.60	2.23	11.13	0.22
						C05 42pg/ml	36.83	25.23	6.16	18.00	0.30

From these results, it can be seen that the proportion of V.delta.1 positive cells in the cultures in which B07, C08, E07 and G04 are present is greater than the OKT3, TS-1 or TS8.2 controls. Thus, the antibodies tested were more efficient at generating and expanding vδ1 positive cells than commercially available antibodies, even when present at low concentrations in culture.

Additional cell markers (including CD 3-CD56+) analysis was also performed on the cells on day 17 of experiment 2 to determine the presence of Natural Killer (NK) cells and CD27-expressing V.delta.1T cells (i.e., CD27+). The results are summarized in table 13.

TABLE 13 percentage of cell composition on day 17-NK cells and CD27+ cells

SEM: standard error of average value

Example 11: functionality of vδ1T cells

The amplified vδ1T cells in the presence of the selected antibodies retained a polyclonal pool of CDR3 regions and also tested for functionality using a SYTOX flow-killing assay. Results are presented for cells obtained using cells with an effector to target (E: T) ratio of 10:1 on day 14 of experiment 1 (FIG. 12A) and cells obtained using cells with E: T ratios of 1:1 and 10:1 on day 17 of experiment 2 (post-freeze thawing) (FIG. 12B).

As can be seen in fig. 12, vδ1 positive cells expanded in the presence of all antibodies effectively lyse target cells, indicating that they are functional even after freezing and thawing the cells.

Example 12: functionality of cells after storage

The functionality of the cells after the storage step of freezing and then thawing was also investigated. A portion of the cells were removed from the culture on day 17 of experiment 2 and frozen. The cells were then thawed and further expanded in culture with IL-15. FIG. 13 shows the total cell count of cultures contacted with B07, C08, E07, G04, or OKT-3 antibodies before freezing after 7 days of cell culture after freeze thawing. All cultures showed the ability to proliferate after storage. Culture was continued until day 42 during which time the total cell count was monitored (results are shown in fig. 14). In cultures previously exposed to the selected antibodies, the total cell number is maintained or increased.

Example 13: binding equivalence study of modified anti-V delta 1 antibodies

ELISA-based antigen titration binding studies were performed to compare the 1245_P02_G04 antibody produced in HEK with its sequence and glycosylation variants produced in CHO. In particular, modifications were made to the framework, allotype, hinge-mediated effector function, asn 297 glycosylation and/or method of manufacture, and were then included in this study. The assay ELISA settings were as follows: the antigen comprises antigen L1 (TRDV 1/TRGV); blocking buffer-2% marvel/PBS; beginning with 5 μg/ml mAb diluted in 1/2 series; antigen-antibody incubation in ELISA plates for-1 hr; washing to remove non-specific binding-3 x PBS-Tween followed by 3 x PBS; DELFIA Eu-labeled anti-human IgG (Perkinelmer; catalog number: 1244-330; 50. Mu.g/ml) diluted with secondary antibody-1/500; after 1 hour incubation, DELFIA enhancing solution (PerkinElmer, used as indicated) was then added; the measurement was performed by Time Resolved Fluorescence (TRF). For antibodies prepared in CHO, the standard expression vectors containing heavy and light chain cassettes were prepared under low endotoxin conditions based on anion exchange chromatography. The DNA concentration was determined by measuring the absorbance at a wavelength of 260 nm. The sequence was verified by sanger sequencing (Sanger sequencing) (up to two sequencing reactions per plasmid, depending on the size of the cDNA). Suspension adapted CHO K1 cells (originally from ATCC and adapted to serum-free growth in suspension culture) were used for manufacturing. Seed cells are grown in chemically defined, animal-component-free, serum-free media. Cells are then transfected with the vector and transfection reagent, and the cells are further grown. The supernatant was harvested by centrifugation and subsequent filtration (0.2 μm filter) and the antibodies were purified using MabSelect ^TM SuRe^TM prior to formulation. To generate an exemplary defucosylated antibody, first the antibody is prepared by von Horsten HH et al (2010) Glycobiology 20 (12): 1607-18 was introduced into the CHO expression platform described above, followed by expression and purification. After manufacture and purification, mAb defucosylation was confirmed by MS-based analysis.

The results of this study are summarized in fig. 15. Wherein the y-axis represents ELISA signal and the x-axis indicates the concentration of V.delta.1 antigen (ug/ml) employed. Antibodies included in this titration study are summarized and more detail is as follows: RSV = anti-RSV control mAb control (manufactured in CHO). G04 =1245_p02_g04 (manufactured in HEK). AD3 = variation G04 (manufactured in CHO). AD4 = hinge modified AD3 (manufactured in CHO). AD3gly = defucosylated AD3 made in engineered CHO. At all titers, equivalent antigen binding was observed for all variants.

Example 14: investigation of binding equivalency of anti-vδ1 antibodies to human germline vδ1 antigen and polymorphic variants thereof

Binding to polymorphic human germline V.delta.1 antigen (SED ID NO: 306) was compared against an IMGT database (see SEQ ID NO: 272) and ELISA-based binding studies were performed to investigate the binding of anti-V.delta.1 antibodies to human germline V.delta.1 antigen. Specifically, the binding and cross-reactivity of antibodies to antigen L1 (containing the coding sequence TRDV/TRGV lines) and L1AV (containing the variation TRDV1/TRGV4 of the TRDV line polymorphism) were compared. The results are presented in fig. 16. The antibodies are shown below: g04 =1245_p02_g04; g04 LAGA = G04 with hinge Fc modification (L235A, G237A EU numbering). E07LAGA =1245_p01_e07 with L235A, G a. C08 LAGA = 1252_p01_c08 with L235A, G237A, d1.3 = control. Serial dilutions were made for each antibody against the antigen, and in all dilutions, equivalent binding to both antigens was observed for each antibody variant. Shows the equivalent binding observed for one example dilution (1 nM antibody) in the series.

Example 15: anti-vδ1 antibody binding increases cytokine secretion by vδ1+ cells

Briefly, all antibodies were diluted to 10 μg/ml and incubated overnight to allow binding of the antibodies to the plate, followed by washing. Skin-derived γδ T cells from two different skin donors were prepared as outlined elsewhere herein (see example 1; specifically, the skin-derived γδ T cell preparation section). These skin cells were then added to tissue culture plates (100,000 cells per well) containing the indicated binding antibodies. The cells were then left for one day, and the supernatant was then harvested and stored at-80 ℃. For cytokine analysis of the supernatant, MSD U-PLEX human assay was used: k151TTK-1, K151UCK-1 (Mesoscale Diagnostics, maryland). Antibodies employed in this study included IgG1 (non-vδ1 binding control), B07 (1245_p01_b07), E07 (1245_p01_e07), G04 (1245_p02_g04; 1245), and C08 (1252_p01_c08). The results of this study are presented in figure 17. Specifically, FIGS. 17 (A) and (B) outline the amounts of TNF- α and IFN- γ detected in the supernatant when skin-derived γδ T cells were applied, and the higher levels observed when the indicated anti-V δ1 antibodies were applied, respectively.

Example 16: anti-vδ1 antibodies increase vδ1+ cell granzyme B levels/activity

Skin-derived γδ T cells were prepared as outlined elsewhere herein (see example 1; skin-derived γδ T cell preparation section). THP-1 cells were first loaded with GranToxiLux probes (cell permeable fluorogenic substrates designed to detect granzyme B activity in target cells) and performed according to the manufacturer's instructions (OncoImmunin, gaithersburg, U.S.). THP-1 cells were then pulsed with antibody at 10 μg/ml and then mixed with skin-derived γδT at a target cell/effector cell ratio of 1:20 as shown in FIG. 18. The co-cultures were then simply centrifuged to ensure rapid formation of the conjugate, then co-cultured for 1 hour, followed by flow analysis according to the GranToxiLux protocol. Antibodies employed in this study included IgG1 (non-vδ1 binding control), B07 (1245_p01_b07), E07 (1245_p01_e07), G04 (1245_p02_g04; 1245), and C08 (1252_p01_c08). The results are presented in figure 18 and highlight the higher levels of granzyme B in target cancer cells observed when the indicated anti-vδ1 antibodies were applied to this vδ1+/THP-1 co-culture model system.

Example 17: anti-vδ1 antibodies confer immune cell modulation and proliferation in human tissues

Human skin punch biopsies (from five different donors) were incubated in culture for 21 days with the indicated antibodies. Skin samples were prepared by removal of subcutaneous fat, etc., as described elsewhere herein (see example 1; gamma delta T cell preparation from skin source). Parallel assay perforations from each donor were then placed on a carbon matrix grid and then placed in the wells of G-REX6 (Wilson Wolf). Each well is filled with complete medium, as described elsewhere herein. To study and compare the effect of different antibodies, these antibodies were added to a working concentration of 100ng/ml on days 0, 7, 14. After 21 days of culture, cells were harvested and analyzed by flow cytometry. The results of the study are presented in fig. 19 and in particular highlight the significant differences in the modulating effect of the different antibodies: sequence ：Vd1TS8.2＝TS8.2(Thermo Fisher);OKT-3(Biolegend);C08 IgG1＝1252_P01_C08;E07＝1245_P01_E07;G04＝1245_P02_G04. from left to right fig. 19 (a) highlights the average number of live pan γδ TCR positive cells observed at the end of culture; results are presented as gating scores (percent mean + standard deviation) of total viable cell populations as analyzed by flow cytometry. For pan γδ content analysis, the flow gating strategy is as follows: single cell > living cell > pan gamma delta antibody (Miltenyi, 130-113-508). FIG. 19 (B) highlights the average number of live V.delta.1+ TCR positive cells observed at the end of the culture; results are presented as percent gating (percent mean + standard deviation) of total viable cell population as analyzed by flow cytometry. For vδ1+ cell content analysis, the flow gating strategy was as follows: single cell > living cell > pan [ gamma ] [ delta ] (Miltenyi, 130-113-508) > V [ delta ] 1 (Miltenyi, 130-100-553). FIG. 19 (C) highlights the number of living biscationic V.delta.1+CD25+ cells observed at the end of the culture; results are presented as gating scores (percent mean + standard deviation) for the total viable cell population. For cd25+vδ1+ cell content analysis, the flow gating strategy was as follows: single cell > living cell > pan [ gamma ] [ delta ] (Miltenyi, 130-113-508) & gt V [ delta ] [ 1 (Miltenyi, 130-100-553) & gt CD25 (Miltenyi, 130-113-286). The combined results of this study summarize the different effects of the antibodies of the invention as described herein relative to the comparison antibodies TS8.2 and OKT 3. While not being bound by this theory, one possibility that TS8.2 or OKT3 exert less favorable effects on vδ1+ cells may be due to the deleterious effects of these comparison molecules on immune cell function over time in the model system.

Example 18: anti-vδ1 antibodies confer modulation and proliferation of immune cells in TIL

Studies were conducted to explore the modulation and proliferation of anti-vδ1 antibodies conferring human Tumor Infiltrating Lymphocytes (TILs). For these studies, human Renal Cell Carcinoma (RCC) tumor biopsies were freshly delivered and treated after receipt. Specifically, the tissue is cut to about 2mm ². Up to 1g of tissue was placed in MILTENYI C tubes along with 4.7mL of RPMI with enzyme from Miltenyi tumor dissociation kit at the manufacturer recommended concentration, which was used at 0.2x concentration except for enzyme R to prevent the relevant cell surface molecules from lysing. The C-tube was placed on GENTLEMACS ^TM Octo ionizer with heater. The procedure 37c_h_tdk_1 was selected for dissociating soft tumors. The digests were then filtered through a 70mM filter to generate a single cell suspension. RPMI containing 10% fbs was added to digests to quench enzyme activity. Cells were washed 2 times with RPMI/10% fbs and resuspended for counting. The derived cells were then seeded into TC wells (24 well G-REX, wilson Wolf) at a density of 2.5x10e6 per well. Cells were then incubated for 18 days with or without cytokines and with or without antibodies. Antibodies included in the study are summarized in figure 20. These include OKT3 (to 50ng/m 1) and 1252_P01_C08, also referred to herein as "C08" (to 500 ng/ml). When included, these antibodies were added on day 0, day 7, day 11, and day 14. During the incubation, the medium was replaced with fresh medium on days 11 and 14. Flow cytometry analysis was performed on day 0 and day 18 to determine fold changes in lymphocyte phenotype and cell number. Cells were first gated on live cd45+ cells and then on demand. In arms comprising recombinant cytokines, these are added as follows. Day 0: IL-4, IFN-gamma, IL-21, IL-1β. Additional IL-15 was added on days 7,11, and 14. Additional IL-21 and IFN-gamma were added on day 7 and day 14, respectively. FIG. 20 (A) shows fold increase in TILvδ1+ cells after 18 days of culture in the presence of C08 or OKT3 with and without cytokine support (CK) under the instruction. These results indicate a significant fold increase in tilvδ1+ cells after application of C08 or comparison OKT3 antibody in the presence of cytokines compared to antibodies or cytokines alone. Fig. 20 (B) shows the increase in total vδ1 cell number after harvest. These results indicate that the number of tilvδ1+ cells increases significantly after incubation with C08 or the comparative OKT3 antibody in the presence of cytokines compared to the antibodies or cytokines used alone. Fig. 20 (C) presents an exemplary gating strategy for cellular flow cytometry analysis. Cells of the live cd45+ cell population were gated against lymphocytes based on the forward and side scatter characteristics of the lymphocytes (not shown), and γδ T cells were then separated from αβ T cells by staining the T cell receptor. Finally, the proportion of vδ1 cells in the total γδ T cell population is determined. Example data for day 18 (+/-1252_p01_c08) are shown for 2 cases: 64.3% of the cells are CD45+, 53.1% of these CD45% cells are γδ+, and 89.7% of these γδ cells are vδ1+. FIG. 20 (D) presents a cell surface phenotype profile of TILvδ1+ cells at harvest. Higher levels of CD69 were observed after incubation with C08 antibody. Figure 20 (E) presents an analysis of tilγδ negative, CD8 positive lymphocyte fractions within the live CD45 positive gate at harvest. In summary, the combined results highlight the modulating effect conferred by the anti-vδ1 antibodies of the present invention described herein on the TIL population.

Example 19: anti-vδ1 antibodies enhance vδ1+ cell-mediated cytotoxicity and diseased cell-specific cytotoxicity

Cytotoxicity/potency assays and studies were performed in a model system comprising a ternary culture of vδ1+ effector cells, THP-1 mononuclear cancer cells, and healthy primary monocytes +/-anti-vδ1 antibodies as described herein (1245_p02_g04; 1245_p01_e07; 1252_p01_c08) and including a control (no mAb or D1.3) as shown in fig. 21. Briefly, all antibodies were diluted to 10 μg/ml in PBS and incubated overnight at 4 ℃ in 384 Kong Chaocheng image assay plates (PERKIN ELMER) to bind the antibodies to the plates, followed by washing with PBS. Healthy control monocytes were isolated from peripheral blood mononuclear cells (PBMC; lonza) by negative selection using magnetically activated cell sorting (MACS; miltenyi Biotec). Monocytes and cultured THP-1 cells (ATCC) were stained with [ 0.5. Mu.M ] CELLTRACE VIOLET and CELLTRACE CFSE viable cell stain for 20 minutes, respectively, and then mixed at a ratio of 1:1. Expanded skin-derived vδ1γδ T cells were isolated from tissue culture flasks and serially diluted to generate a range of effector to target cell ratios (E: T) and then added to THP 1: monocyte cell suspensions. The cell suspension was seeded into 384 well assay plates to give a final cell seeding density of 1,000 THP-1 cells per well, 1,000 monocytes per well and a series of γδ T cells (highest E: T ratio of 60:1). To determine the number of live THP-1 and healthy control monocytes after 24 hours, nine fields of view were captured at10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on size, morphology, texture and intensity of viable cell staining. The results are presented in fig. 21. FIG. 21 (A) presents THP-1 and monocyte numbers after 24 hours of ternary co-culture with γδ T cells in the presence of the indicated plate-bound mAbs or controls. Cell numbers were calculated using a high content confocal microscope for live cell imaging. FIG. 21 (B) presents a bar chart representation designed to highlight the window between lesion-specific killing and non-lesion healthy cell retention after 24 hours of co-culture at the highest E:T ratio (60:1): left bar graph; fold increase in diseased cell killing (THP-1) versus non-diseased cells (primary human monocytes). Right side bar graph; the same data, but expressed as percent of enhanced killing compared to control. Figure 21 (C) presents tabulated results summarizing the percent improvement in efficacy of vδ1γδ T cells to kill THP-1 target cells in the presence of vδ mAb, calculated according to figure (a), compared to no mAb control. Figure 21 (D) presents tabulated results of EC50 values calculated according to figure (a), expressed as the number of γδ T cells required to confer 50% thp-1 cell killing. The combined results and findings as outlined in fig. 21 highlight the ability of the antibodies described herein to enhance cytotoxicity and diseased cell specificity of vδ1+ cells.

Example 20: multispecific antibodies potentiate vδ1+ effector cell-mediated cytotoxicity; targeting tissue-centric disease-associated antigens

Cytotoxicity/potency assay studies were performed to explore the effect of multispecific antibodies on vδ1+ effector cell and a-431 cancer cell co-cultures. A-431 (EGFR ⁺⁺; ATCC) target cells were seeded at 1,000 cells/well in 384 well imaging plates (PERKIN ELMER) and incubated overnight in DMEM (10% FCS) at 37 ℃. The indicated antibodies and multispecific antibodies were diluted to 10 μg/ml and added to the assay plate (2 μg/ml final assay concentration). Amplified skin-derived V.delta.1. Gamma. Delta. T cells were isolated from tissue culture flasks and serially diluted to give a range of E:T ratios (highest E:T ratio of 60:1) and then added to assay plates. A-431 cells were incubated with V.delta.1. Gamma. Delta. T cells in the presence of antibodies or controls in 5% CO ₂ at 30 ℃. After 24 hours incubation, hoechst 33342 (ThermoFisher) was added to the stained cells (final 2 μm). To determine the number of viable a-431 cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on size, morphology, texture and intensity of viable cell staining. Effector/target (E: T) time course studies were performed to determine the ET ratio at which 50% of target cells were killed in the model system of +/-shown controls, antibodies and multispecific antibodies. The results are presented in fig. 22.

First, FIG. 22 (A-D) presents exemplary co-culture results, wherein vδ1+/A-431 co-culture is +/-a study of multispecific antibodies comprising an anti-vδ1×anti-TAA (EGFR) bispecific binding moiety, wherein the anti-vδ1VL+VH binding domain (for the first target) is combined with the CH1-CH2-CH3 domain of the anti-EGFR binding moiety (for the second target). Controls and comparisons employed as shown; from left to right: no mAb = no added antibody; d1.3 Control =d1.3; d1.3igg LAGA = d1.3+l235A, G a; D1.3FS1-67 = D1.3 variable domain and EGFR binding constant domain plus L235A, G a; cetuximab (internally generated). More specifically, fig. 22 (a) presents the results of five hours of co-incubation with the control, the comparator, and the following test articles described above: c08-LAGA = 1252_p01_c08 with L235A, G237A; c08 FS1-67 = 1252_p01_c08 containing L235A, G237A in combination with an EGFR binding domain. Fig. 22 (B) presents equivalent data for five hours of co-incubation with the control, comparator, and test article described above: g04-LAGA =1245_p02_g04 with L235A, G237A; g04 FS1-67 = 1245_p02_g04 containing L235A, G237A in combination with an EGFR binding domain. Fig. 22 (C) presents equivalent data for five hours of co-incubation with control, comparator and test article below: e07-LAGA =1245_p01_e07 with L235A, G237A; e07 FS1-67 = 1245_p01_e07 containing L235A, G237A in combination with an EGFR binding domain. Fig. 22 (D) presents a table summarizing the percent improvement in cytotoxicity of vδ1γδ T cells in the presence of control, comparator and test article over 5 hours, 12 hours and 24 hours. When an antibody or antigen binding fragment thereof as described herein is presented in a multispecific format, an enhancement of greater than 450% can be observed.

Second, FIG. 22 (E-H) presents exemplary results in which vδ1+/A-431 co-cultivation is +/-a study of multispecific antibodies comprising anti-vδ1×anti-TAA (EGFR) bispecific binding moieties, where the anti-vδ1 binding domain (for the first target) comprises a full length antibody (VH-CH 1-CH2-CH 3/VL-CL) followed by combination with an anti-EGFR scFv binding moiety (for the second target). Controls and comparisons employed as shown; from left to right: no mAb = no added antibody; d1.3 Control; d1.3 IgG LAGA = d1.3+l235A, G a; d1.3 LAGA cetuximab = D1.3 with L235A, G237A plus a C-terminal cetuximab-derived scFv; cetuximab (internally generated). More specifically, fig. 22 (E) presents five hour co-cultures with the controls, comparisons, and test articles described above: c08-LAGA = 1252_p01_c08 with L235A, G237A; c08 LAGA cetuximab = 1252_p01_c08 with L235A, G237A and with a C-terminal cetuximab-derived scFv. Fig. 22 (F) presents five hour co-cultures with the controls, comparisons, and test articles described above: g04-LAGA =1245_p02_g04 with L235A, G237A; g04 LAGA cetuximab = 1245_p02_g04 with L235A, G237A and with a C-terminal cetuximab-derived scFv. Fig. 22 (G) presents five hour co-cultures with controls, comparisons and the following test articles: e07-LAGA =1245_p01_e07 with L235A, G237A; e07 LAGA cetuximab = 1245_p01_e07 with L235A, G237A and with a C-terminal cetuximab-derived scFv. Figure 22 (H) presents a table summarizing the percentage improvement in potency of vδ1γδ T cells in the presence of controls, comparisons and test articles over 5 hours, 12 hours and 24 hours. When an antibody or antigen binding fragment thereof as described herein is presented in a multispecific format, an enhancement of greater than 300% can be observed.

Third, fig. 22 (I and J) outlines an exemplary alternative method of representing data. Specifically shown is the percentage increase in cytotoxicity of vδ1+ effector cells against egfr+ cells at the 24 hour time point for the multispecific antibodies E07 FS1-67 (I) or C08 FS1-67 (J) relative to all components and comparisons shown.

Example 21: the multispecific antibodies enhance vδ1+ mediated cytotoxicity and diseased cell-specific cytotoxicity; targeting hematopoietic-centered disease-associated antigens

Cytotoxicity/potency assays and studies were performed in a model system comprising vδ1+ effector cells and Raji cancer cells as well as healthy primary monocytes +/-a ternary culture comprising multispecific antibodies to vδ1×anti-TAA (CD 19) multispecific antibodies to determine whether a dual specificity format tumor-associated antigen (TAA) linked vδ1 monoclonal antibody can enhance vδ1γδ T cell killing of a particular target cell. Specifically, raji cells (CD19++; ATCC) were incubated with V.delta.1γδT cells in the presence of V.delta.1×CD19 multispecific antibodies. All antibodies were diluted to 4. Mu.g/ml (final assay concentration 1. Mu.g/ml) and added to 384 well imaging plates (PERKIN ELMER). Amplified skin-derived V.delta.1. Gamma. Delta. T cells were isolated from tissue culture flasks and serially diluted to obtain a range of effector to target cell ratios (E: T). Raji cells were stained with [ 0.5. Mu.M ] CELLTRACE FAR RED and then mixed with titrated V.delta.1γ.delta.T cells at a 1:1 ratio. The cell suspension was seeded into 384 well assay plates to give a final cell seeding density of 1,000 Raji cells per well and a series of γδ T cells (highest E: T ratio 30: 1). To determine the number of viable Raji cells after 24 hours, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on size, morphology, texture and intensity of viable cell staining. The results are captured in fig. 23. The antibodies and comparisons used therein are shown. Specifically, RSV IgG = motuzumab non-binding control, g04 = 1245_p02_g04; e07 =1245_p01_e07; D1.3VHVL = D1.3HEL with heavy chain C-terminal anti-CD 19 scFv (scFv binding module employed see SEQ ID NO: 314); g04 Vhvl=1245_p02_g04lag with heavy chain C-terminal anti-CD 19 scFv (SEQ ID NO:315 or SEQ ID NO:421 and SEQ ID NO: 422); e07 VHVL= 1245_P01_E07 LAGA (SEQ ID NO:316, SEQ ID NO:423 and SEQ ID NO: 424) with heavy chain C-terminal anti-CD 19 scFv. FIG. 23 (A) is a table summarizing (i) calculated EC50 expressed as the number of γδ T cells or E:T ratio required to induce 50% Raji cell killing, and (ii) percent improvement in EC50 over mAb-free control. FIG. 23 (B) is a bar graph showing the percent improvement in the ability of γδ T cells to lyse 50% Raji target cells in the presence of a V.delta.1-CD 19 multispecific antibody.

Example 22 mammalian display

Two clones were selected for affinity maturation. The preparation and characterization of affinity matured clones derived from clone ADT1-4 (G04) and clone ADT1-7 (E07) will now be presented.

Affinity maturation of human anti-V delta 1 monoclonal antibodies

Phage display for the production of the parent anti-vδ1 monoclonal antibody produced antibodies with affinities in the range of 12nM-1 μm, as described above. Then, parent antibody clone ADT1-4 (G04) and clone ADT1-7 (E07) were affinity matured in vitro, unexpectedly achieving 100-fold improved affinity, and thus excellent target engagement. In vitro affinity maturation of parent antibodies is achieved by a two-step process: parental antibody sequences were diversified using targeted CDR3 mutagenesis, followed by selective enrichment of affinity-improved antibodies using phage and mammalian display platforms. VH and VL CDR32 mer libraries of clones ADT1-4 and ADT1-7 were created using Kunkel mutagenesis (Kunkel et al, 1987; sidhu and Weiss, 2004) and RCA amplification. All single and double amino acid substitutions at specified positions in VH and VL CDR3 were combined using Agilent primer synthesis techniques. The number of different amino acids to be incorporated at a particular position is specified. Cysteine and methionine are omitted. The changes made in the cloned CDRs are summarized in table 14 below.

TABLE 14 CDR3 position and residue specified changes

The size of each library is shown in table 15.

TABLE 15 library size

ADT1-7	5.4×109
		ADT1-4	5.5×109

Mutagenesis generation of VH and VL CDR3 libraries

The mutagenic library is prepared from a suitable culture volume covered with a suitable excess of library members. Affinity maturation was performed using solution phase selection using phage display technology as described by Schofield et al (2007). Selection was performed in solution using human and cynomolgus monkey antigens (human antigens include human TRDV sequences, cynomolgus monkey antigens include SEQ ID NO:308 minus leader sequences). To isolate the binders from a library of mutants with higher affinity, the antigen concentration is controlled to make a series of increasingly stringent selections. Several rounds of phage display selection were performed with human and cynomolgus monkey antigens for each clone as presented in figure 24. For the ADT1-4 lineage, the antigen concentration range used in the selection was: human antigen 100nM-10pM, cynomolgus monkey antigen 100nM and 10nM. For the ADT1-7 lineage, human antigen concentrations of 10nM to 1pM were used, while cynomolgus monkeys were 100nM to 100pM. FIG. 24 shows phage selection rounds for ADT1-7 library (iA), ADT1-4 library (iB) and ADT1-7 library (iC) using a selection strategy for isolating cynomolgus cross-reactive binders.

The progress of selection was assessed using a polyclonal phage ELISA. Human antigen (DV 1/GV 4) and Cyno antigen (DV 1/GV 76) were coated at 150 ng/well, 50. Mu.L/well overnight. Cynomolgus monkey antigens DV2/GV76 (which does not contain SEQ ID: 308) and HSA were used as controls. Only the output of the ADT4-1 library cross-reacted with both human and cynomolgus monkey DV1 antigen. The output of ADT1-7 is only responsive to human antigens. The selected outputs were further characterized by monoclonal phage ELISA and the summary is presented in table 16. The light grey and dark grey boxes indicate selection with human or cynomolgus monkey antigen, respectively. Arrows indicate the percentage of clones classified as human or cynomolgus VD1 conjugates (upward pointing high and downward pointing low).

TABLE 16 ELISA summary of monoclonal phages

The sequence diversity is summarized in table 17. The light grey and dark grey boxes indicate selection with human or cynomolgus monkey antigen, respectively. Arrows indicate the level of diversity (high upwards and low downwards).

TABLE 17 sequencing summary of monoclonal phages

Generation of IgG mammalian display libraries

To form the final library for mammalian display, the following ADT1-4 selections were pooled:

SEL2409, 2412 and 2413, creating ADT1-4 library 1 (human)

SEL 2418, 2420 and 2415, create ADT1-4 library 2 (cynomolgus monkey)

SEL2370 and 2404, create ADT1-7 libraries.

The pool then progresses to mammalian display. A population of single chain variable fragment antibodies (scFv) is converted in its entirety to IgG format, maintaining the original Variable Heavy (VH) and Variable Light (VL) pairing. The IgG format antibody is then cloned into a mammalian display donor vector, which is co-transfected with a plasmid encoding a TALE nuclease pair, such that the nuclease directs the integration of the antibody gene at a single chromosomal site. Mammalian display antibody libraries covering phage output diversity (> 10 ⁶ clones) were created in HEK293 cells. Stable cell populations expressing antibodies on the cell surface were selected by adding blasticidin (dpt) 2 days after transfection. Cells expressing antibodies on the cell surface were enriched by Magnetic Activated Cell Sorting (MACS) sorting (7 dpt); cells were labeled with anti-Fc-PE, then anti-PE microbeads, and sorted using Midi MACS magnet (Miltenyi Biotec) and LS column. These cell populations progress to selection.

Selection of mature antibodies by mammalian display

After enrichment of antibody expressing cells with MACS, two strategies were used to identify TRDV binders. The main strategy involves two-color fluorescence sorting based on Fc expression and antigen binding. Another involves two-color fluorescence sorting based on cynomolgus monkey and human antigen binding to maximize the chance of separating high affinity cross-reactive conjugates. Fig. 25 presents a schematic view of this process.

Sequence, specificity, affinity ordering and characterization

Genomic DNA was extracted from eight different sorted populations. The DNA encoding the selected IgG was amplified and cloned into a soluble IgG1 expression vector. A total of 1472 clones were selected from 8 different selections. pDNA was transfected into Expi293 cells. Supernatants were harvested 5 days post-transfection and the affinity of the expressed antibodies for binding to human and cynomolgus TRDV1 was ranked in a capture ELISA. A total of 93 anti-DV 1 antibodies were selected from each selection strand for sequence and SPR dissociation rate analysis. Furthermore, these clones were checked for binding by direct ELISA to: human TRDV1, polymorphic human TRDV1 (A.fwdarw.V), human TRDV, cynomolgus TRDV1 and BSA.

Example 23 binding affinity to human cynomolgus monkey antigen

Binding to recombinant antigen expressed on cells and vδ1 TCR: ADT1-4 lineage

Studies were performed to explore the binding of anti-vδ1 antibodies to their target antigens. Binding of anti-vδ1 mAb to vδ1 TCR antigen was tested by ELISA. 1ug of human antigen or 1ug of cynomolgus monkey antigen per well was immobilized on a 96-well immunoassay plate (SLS# 475904) and then blocked with BSA to prevent non-specific binding. 1.3pmol (20 ng) of each mAb was added and incubated for 1 hour at room temperature. Binding of mAb to antigen was detected by measuring absorbance at 450nM using protein A-HRP (Abcam#Ab7456) and TMB substrate (Fisher# 12750000) and stop solution (bioleged# 423001). The parental control was included as a positive control for assay and variability between plates. Hits were identified as those mabs with higher absorbance readings than the parent mAb. FIG. 28A shows the fold increase in absorbance of mature ADT1-4 clones when 1.3pmol of antibody was conjugated to human v61 antigen as compared to the absorbance of parent ADT1-4 conjugated. FIG. 28B shows the fold increase in absorbance of mature ADT1-4 clones when 1.3pmol of antibody was conjugated to cynomolgus monkey vδ1 antigen as compared to the absorbance of parent ADT1-4 binding.

MAb binding to endogenous vδ1 TCR was tested using flow cytometry. Skin V.delta.1 cells (donor ATS006; ADT expanded E0000113) or PEER V.delta.1 cell lines were seeded into 96-well round bottom plates at 3X 10. Sup.5 cells/well and resuspended in 50ul of FACS buffer (V/V: 2% FCS in PBS, 0.1% sodium azide and 1mM EDTA) containing 3ug/ml test mAb for 15 min at 4 ℃. The cells were pelleted and anti-hmIgG-APC secondary antibody (Miltenyi # 130-119-772) was added at 1/100 in FACS buffer and incubated for an additional 20 min at 4 ℃. Cells were washed and fixed in CellFix (BD # 340181) and analyzed by flow cytometry. The vδ1 phenotype% and the average fluorescence intensity were calculated (Inivai Technologies, flowlogicv7.2). As positive control, the parent antibody was included. FIG. 28C shows the fold increase in mean fluorescence intensity of mature ADT1-4 clones binding to primary skin-derived human V.delta.1 cells compared to parental ADT 1-4. FIG. 28D shows the fold increase in mean fluorescence intensity of mature ADT1-4 clones binding transformed PEER V delta 1 cell lines compared to parental ADT 1-4.

Binding to recombinant antigen expressed on cells and vδ1 TCR: ADT1-7 lineage:

Studies were performed to explore the binding of ADT1-7 mature anti-vδ1 antibodies to their target antigens. Binding of anti-vδ1 mAb to vδ1 TCR antigen was tested by ELISA. 1ug of antigen per well was immobilized on a 96-well immunoassay plate (SLS# 475904) and then blocked with BSA to prevent non-specific binding. Titration of 6.7pmol (100 ng), 1.3pmol (20 ng) and 0.27pmol (4 ng) mAb was added and incubated for 1 hour at room temperature. Binding of mAb to antigen was detected by measuring absorbance at 450nM using protein A-HRP (Abcam#Ab7456) and TMB substrate (Fisher# 12750000) and stop solution (bioleged# 423001). The parental control was included as a positive control for assay and variability between plates. Hits were identified as those mabs with higher absorbance readings than the parent mAb. FIG. 29A shows fold increase in absorbance of mature ADT1-4 clones when 1.3pmol of antibody was bound to human vδ1 antigen as compared to the absorbance of parent ADT1-4 binding.

MAb binding to endogenous vδ1 TCR was tested using flow cytometry. Skin V.delta.1 cells (donor ATS006; ADT expanded E0000113) or PEER V.delta.1 cell lines were seeded into 96-well round bottom plates at 3X 10. Sup.5 cells/well and resuspended in 50ul of FACS buffer (V/V: 2% FCS, 0.1% sodium azide and 1mM EDTA in PBS) containing 3ug/ml test mAb for 15 min at 4 ℃. The cells were pelleted and anti-hmIgG-APC secondary antibody (Miltenyi # 130-119-772) was added at 1/100 in FACS buffer and incubated for an additional 20 min at 4 ℃. Cells were washed and fixed in CellFix (BD # 340181) and analyzed by flow cytometry. The vδ1 phenotype% and the average fluorescence intensity were calculated (Inivai Technologies, flowlogicv7.2). As positive control, the parent antibody was included. FIG. 29B shows the fold increase in mean fluorescence intensity of mature ADT1-4 clones binding to primary skin-derived human V.delta.1 cells compared to parental ADT 1-4. FIG. 29C shows the fold increase in mean fluorescence intensity of mature ADT1-7 clones binding transformed PEER V delta 1 cell lines compared to parental ADT 1-7.

Fold improvement over parental clones

Dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA)

To confirm the fold improvement in binding of affinity matured mAbs to human and cynomolgus monkey antigens, DELFIA immunoassays were performed by directly coating the antigen onto plates (3. Mu.g/mL antigen in 50. Mu.L PBS overnight at 4 ℃ (Nunc# 437111.) for detection, DELFIA Eu-N1 anti-human IgG (PERKIN ELMER #1244-330) was used as secondary antibody, diluted 1/500 in 50. Mu.L 3% MPBS (PBS+3% (w/V) skimmed milk powder) and developed using 50. Mu.L DELFIA enhancement solution (PERKIN ELMER #4001-0010).

Affinity sequencing was performed on the antibodies of interest using a DELFIA immunoassay, in which the antibodies were captured via protein G coated on a plate, and 0.4nM of human soluble biotinylated L1 (DV 1-GV 4) antigen and 10nM of cynomolgus monkey antigen DV1/GV77 (3 MPBS) were added. For detection, 50. Mu.L of streptavidin-Eu (1:500 in assay buffer, perkin Elmer) was used and the signal was developed with DELFIA enhancement solution. D1.3hIgG1 (described in England et al (1999) J.Immunol.162:2129-2136) was used as a negative control. The results are provided in fig. 30A to C.

SPR analysis-IgG capture

SPR analysis was used to compare KD values of affinity matured clones with that of the parental clones. The instrument is used: MASS-2 (Sierra Sensors); and (3) a chip: amine high capacity (Sierra Sensors); the running buffer was pbs+0.02% tween 20. Experiments were performed at room temperature or 37℃and G protein was coupled to the chip. Antigen was flowed through cells in a series of dilutions, human DV1-GV4 was 50nM to 0.2nM, and cynomolgus monkey DV1-77 was 100nM to 1nM.120 seconds association, 600 seconds dissociation, 50 μl/min flow rate, regeneration with 10mM glycine pH 1.5, kinetic fitting was performed according to langmuir 1:1 combination using the software Sierra Analyzer. The results are shown in fig. 31A to C.

Binding affinity assay (KD, by SPR)

Binding affinity of vδ1 monoclonal antibodies to vδ1 antigen

Binding affinity of the antibodies to the target (i.e., vδ1 chain of γδtcr) was established by SPR analysis using Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Antigen (e.g., L1 (DV 1-GV 4) was flowed through cells in a 1:3 dilution series of 300nM to 3.7nM with 180 seconds of association, 600 seconds of dissociation, a flow rate of 25. Mu.L/min, running buffer PBS+0.05% Tween 20. All experiments were performed at room temperature.

Human antigens

Affinity matured vδ1 mAb of either parent showed greatly enhanced affinity for human vδ1 antigen as determined by Surface Plasmon Resonance (SPR) analysis (fig. 32). (A, B) Surface Plasmon Resonance (SPR) analysis was performed using ADT1-4 (A) or ADT1-7 (B) parent mAbs and affinity matured derivatives thereof to determine binding affinity to human V.delta.1 antigen. The binding interactions of vδ1 and affinity matured mabs were modeled according to langmuir 1:1 binding. The (C, D) table summarizes the equilibrium dissociation constants (KD) of the vδ1-targeted antibodies, derived from the sensorgrams shown in (a) and (B). Data are presented as the average of two replicates performed on two different SPR instruments. The bar graph (E, F) shows the fold increase in equilibrium dissociation constant (KD) for the V.delta.1 antigen for affinity matured derivatives compared to their parent ADT1-4 or ADT 1-7.

Conclusion: the data indicate that affinity-matured derivatives of ADT1-4 and ADT1-7 exhibit significantly higher affinity for human vδ1 antigen than the parent antibody.

Cynomolgus monkey antigen

Surface plasmon resonance analysis showed that affinity of affinity-matured vδ1 mAb of ADT1-4 lineage was greatly enhanced for cynomolgus vδ1 antigen compared to the parent antibody (fig. 33). (A) Surface Plasmon Resonance (SPR) analysis was performed using the ADT1-4 parent antibody and affinity matured derivatives to determine binding affinity to cynomolgus monkey vδ1 antigen. The binding interactions of vδ1 and affinity matured mabs were modeled according to langmuir 1:1 binding. (B) The table summarizes the equilibrium dissociation constants (KD) of the vδ1-targeted antibodies, derived from the sensorgrams shown in (a). Data are presented as the average of two replicates performed on two different SPR instruments. The results are shown in fig. 33A and 33B.

Conclusion: this data shows that affinity-matured derivatives of ADT1-4 exhibit significantly higher affinity for cynomolgus monkey vδ1 antigen than the parent antibody.

Binding affinity to cell surface vδ1 TCR (EC 50 binding to cell surface vδ1)

Affinity matured vδ1 mAb exhibited greatly enhanced affinity for vδ1-positive γδt cells without significant binding to vδ1-deficient cells, as shown in fig. 34A-D. Binding levels of (A, B) vδ1 mAb to two γδ T cell donors ATS006 (a) and TS164 (B). γδ T cells were stained with different concentrations of vδ1 mAb, followed by fluorescent anti-human IgG detection antibody (xxx). All incubation steps were performed at 4 ℃ and mAb binding was determined using flow cytometry to measure median fluorescence levels. GRAPHPAD PRISM 9 was used to fit a log-four parameter dose-response curve. (C) Bars represent the average EC50 for binding of ADT1-4 and ADT1-7 clones to vδ1-positive γδ T cells, expressed as the average of two donors. (D) The table summarizes the EC50 plotted in (a) and (B), as well as the vδ1 negative cell types, including HEK293A, raji cells and various leukocyte subpopulations within primary blood mononuclear cells. For vδ1 positive γδ T cells, the data are expressed as the average of two donors.

Conclusion: this data shows that affinity-matured derivatives of ADT1-4 and ADT1-7 have significantly higher affinity for vδ1-positive γδ T cells than their parent mAb, while not binding to vδ1-negative cells HEK293T, raji or CD8, CD4, NK, CD19 or monocytes within PBMCs.

Target cell binding of vδ1 monoclonal antibodies

A series of target cells were assayed to determine the specificity and affinity of vδ1 mAb binding. This includes expanded vδ1γδ T cells, HEK293T cells, raji cells and multiple leukocyte subpopulations within human primary blood mononuclear cells of skin origin. Adherent or semi-adherent cells (skin-derived vδ1γδ T cells, HEK 293T) were isolated from tissue culture flasks and resuspended in PBS. Similarly, non-adherent cell types (Raji, PBMC) were harvested and resuspended in PBS. Cells were seeded in v-bottom 96-well plates at a final density of 100,000 cells per well. The cells were centrifuged and the cell pellet resuspended in FcR blocking reagent according to the manufacturer's instructions and incubated for 20 minutes at 4 ℃ before further washing. Vδ1 mAb, anti-RSV IgG control and anti-CD 3 OKT3 were diluted to 500nM in PBS and serially diluted 1:5 to 6.4pM in PBS and added to cells, followed by incubation at 4 ℃ for 20 minutes. To determine the number of mAbs bound to the cell surface, cells were then stained with murine anti-human IgG secondary antibodies conjugated to APC (dilution: 1:100). For vδ1γδ T cells, HEK293T and Raji, cells were also stained with reactive dye only. Conjugated antibodies to CD4, CD8, CD56, CD11B and CD19 (all 1:100) were also included for PBMC, so that αβ subpopulations, NK cells, B cells and monocytes could be distinguished. After incubation at 4 ℃ for 20 minutes, the cells were washed twice and fluorescence was measured using MACSQuant. The IC50 is shown in fig. 34D.

KD value summary of ADT1-4 lineage (human and cynomolgus monkey)

The following table provides KD values (binding to human TRDV1 and cynomolgus monkey TRDV 1) for 24 clones in ADT1-4 lineage and for the ADT1-4 parental clone (G04):

KD value summary of ADT1-7 lineage (human only)

The following table provides KD values (binding to human TRDV1 only) for 11 clones in ADT1-7 lineage and for the ADT1-7 parental clone (E07):

Clone ID	KD[M]	KD(nM)
			ADT1-7	1.24E-08	12.4nM
ADT1-7-10	1.30E-09	1.30nM
			ADT1-7-15	1.42E-09	1.42nM
ADT1-7-17	1.26E-09	1.26nM
			ADT1-7-18	1.61E-09	1.61nM
ADT1-7-19	1.19E-09	1.19nM
			ADT1-7-20	1.17E-09	1.17nM
ADT1-7-22	1.18E-09	1.18nM
			ADT1-7-23	1.29E-09	1.29nM
ADT1-7-42	1.51E-09	1.51nM
			ADT1-7-3	8.31E-10	0.831nM
ADT1-7-61	5.57E-10	0.557nM

Summary of pharmacological data for preferred members of the ADT1-4 lineage

* ADT1-4 results did not reach full saturation in this assay, so the data was extrapolated

Summary of pharmacological data for preferred members of the ADT1-7 lineage

* ADT1-7-61 assay represents only one donor

* ADT1-7 results did not reach full saturation in this assay, so the data was extrapolated

Example 24 functional characterization

Effect of V.delta.1 monoclonal antibodies on TCR Down-regulation

The ability of ADT mabs to bind to and thereby down-regulate γδ T cell receptors was assessed by measuring TCR expression. Skin γδ T cells (from donors ATS006 and TS 164) were seeded at 3×10≡5 cells/ml in 96 well round bottom plates in γδ medium with increasing concentrations of test mAb (ranging from 0.00067 to 67 nM) or corresponding isotype control (hIgG 1, RSV) at the highest concentration (67 nM) diluted in PBS. Cells were incubated in a humidified CO2 chamber at 37℃for 2 hours. Cells were washed and stained for dead cells (Thermo Fisher # 15580607) and their VD1 TCR (Miltenyi # 130-117-697) at 4℃for 30 minutes. Cells were washed in FACS buffer and resuspended in Cell Fix (BD sciences # 340181) and then incubated overnight in the dark at 4 ℃. The following day VD1 TCR expression levels as determined by Median Fluorescence Intensity (MFI) were measured by flow cytometry using MACS Quant Analyzer.

The results are shown in fig. 35, which shows that affinity-matured vδ1 mAb binds and down-regulates vδ1 TCR more efficiently than its parent clone and OKT3 in the soluble assay format (a and B). This compelling finding underscores that the methods employed herein involving the use of high affinity vδ1 mAb to down-regulate/activate vδ1 receptor-containing TCR/CD3 complexes are not only much more selective, but are also far more efficient than traditional CD3 targeting methods. γδ cells were incubated with vδ1 mAb and OKT3 for 2 hours, then vδ1 TCR was stained with a-vδ1 TCR-PE-Vio770, and γδ TCR expression was quantified. MFI for a-vδ1 staining was obtained using flow cytometry and calculated from TCR-positive gates, and then MFI% was calculated from MFI of TCR-positive, untreated γδ cells. The upper panel shows data from ADT1-4 lineage and the lower panel shows data from ADT1-7 lineage. The bar graph (C) shows the IC50 values calculated according to (a). Bars (D and E) represent IC50 values in (B) presented as fold improvement over the parent ADT1-4 (left) and ADT1-7 (right). (F) Table shows the IC50 values (in nM) from (B) and the percent improvement calculated from the parents of each of ADT1-4 (top) and ADT1-7 (bottom). (G) Table showing IC50 values from (B) and percent improvement calculated from the parents of each of ADT1-4 (top) and ADT1-7 (bottom).

The effect of V.delta.1 monoclonal antibodies on gamma.delta.activation, measured by CD107a expression

The ability of ADT mabs to bind to and activate vd1γδ cells was assessed by measuring CD107a expression. Skin γδ T cells (from donor ATS 006) were seeded at 6×10≡5 cells/ml in 96 well round bottom plates in γδ medium with 1.2X10≡6 cells/ml THP-1 cells (ATCC-TIB-202) and an increased concentration of test mAb (ranging from 0.00067 to 67 nM) or the highest concentration (67 nM) of the corresponding isotype control (hIgG 1, RSV) diluted in PBS. Cells were incubated in a humidified CO2 chamber at 37℃for 2 hours. Cells were washed and stained for dead cells (Thermo Fisher # 15580607), VD1 TCR (Miltenyi # 130-117-697) and aCD107a (Miltenyi # 130-112-610) at 4℃for 30 minutes. Cells were washed in FACS buffer and then incubated overnight at 4 ℃ in the dark. The following day VD1 TCR and CD107a expression levels, as determined by Median Fluorescence Intensity (MFI), were measured by flow cytometry using MACS Quant Analyzer.

The results are shown in fig. 36, which shows that affinity matured vδ1 mAb induced activation only in the presence of target cells. (A and B) γδ cells were incubated with ADT1-4-2 alone (upper panel) and with THP-1 cells (lower panel) at a target cell to effector cell ratio of 1:2 for 2 hours, then stained with a-V δ1TCR-PE-Vio770 and a-CD107a-VioBlue for V δ1TCR and CD107a, respectively, and then γδ TCR expression (black) and CD107a expression (grey) were quantified. MFI for vδ1 staining was obtained using flow cytometry and calculated from TCR-positive gates, and then MFI% was calculated from MFI of TCR-positive, untreated and non-co-cultured γδ cells. CD107 a-stained MFI was obtained using flow cytometry and calculated from the 'non-THP-1' gate, and then MFI% was calculated from CD107a MFI of untreated, non-co-cultured γδ cells. (C and D) γδ cells were incubated with ADT1-7-3 alone (upper panel) and with THP-1 cells (lower panel) at a target cell to effector cell ratio of 1:2 for 2 hours, then V.delta.1 TCR and CD107a were stained with a-V.delta.1 TCR-PE-Vio770 and a-CD107a-VioBlue, respectively, and γδ TCR expression (black) and CD107a expression (grey) were quantified. MFI for vδ1 staining was obtained using flow cytometry and calculated from TCR-positive gates, and then MFI% was calculated from MFI of TCR-positive, untreated and non-co-cultured γδ cells. CD107 a-stained MFI was obtained using flow cytometry and calculated from the 'non-THP-1' gate, and then MFI% was calculated from CD107a MFI of untreated, non-co-cultured γδ cells. (E) Table shows the percentage increase in γδ CD107a expression of co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, non-co-cultured γδ cells. (F) Table shows the percentage increase in γδ CD107a expression of co-cultured cells treated with the highest concentration vδ1 mAb compared to untreated, co-cultured and non-co-cultured γδ cells.

The effect of V.delta.1 monoclonal antibodies on gamma.delta.activation, measured by CD25 expression

The ability of ADT mabs to bind to and activate vd1γδ cells was assessed by measuring CD25 expression. Skin γδ T cells (from donor ATS 006) were seeded at 6×10≡5 cells/ml in 96 well round bottom plates in γδ medium with 1.2X10≡6 cells/ml THP-1 cells (ATCC-TIB-202) and an increased concentration of test mAb (ranging from 0.00067 to 67 nM) or the highest concentration (67 nM) of the corresponding isotype control (hIgG 1, RSV) diluted in PBS. Cells were incubated in a humidified CO2 chamber at 37℃for 24 hours. Cells were washed and stained for dead cells (Thermo Fisher # 15580607), VD1 TCR (Miltenyi # 130-117-697) and CD25 (Miltenyi # 130-113-280) at 4℃for 30 minutes. Cells were washed in FACS buffer and then incubated overnight at 4 ℃ in the dark. The following day VD1 TCR and CD107a expression levels, as determined by Median Fluorescence Intensity (MFI), were measured by flow cytometry using MACS Quant Analyzer.

The results are shown in fig. 37, which shows that affinity matured vδ1 mAb induced activation only in the presence of target cells. (A and B) γδ cells were incubated with RSV alone (added at 67 nM) and ADT1-4-2 (upper panel) and with THP-1 cells (lower panel) at a target cell to effector cell ratio of 1:2 for 24 hours, then stained with a-V δ1 TCR-PE-Vio770 and a-CD25-VioBlue for V δ1 TCR and CD25, respectively, and then γδ TCR expression (black) and CD25 expression (grey) were quantified. MFI for vδ1 staining was obtained using flow cytometry and calculated from TCR-positive gates, and then MFI% was calculated from MFI of TCR-positive, untreated and non-co-cultured γδ cells. CD25 stained MFI was obtained using flow cytometry and calculated from the 'non-THP-1' gate, and then MFI% was calculated from CD25 MFI of untreated, non-co-cultured γδ cells. (C and D) γδ cells were incubated with RSV alone (added at 67 nM) and ADT1-7-3 (upper panel) and with THP-1 cells (lower panel) at a target cell to effector cell ratio of 1:2 for 2 hours, then stained with a-V δ1 TCR-PE-Vio770 and a-CD25-VioBlue for V δ1 TCR and CD25, respectively, and then γδ TCR expression (black) and CD25 expression (grey) were quantified. MFI for vδ1 staining was obtained using flow cytometry and calculated from TCR-positive gates, and then MFI% was calculated from MFI of TCR-positive, untreated and non-co-cultured γδ cells. CD25 stained MFI was obtained using flow cytometry and calculated from the 'non-THP-1' gate, and then MFI% was calculated from CD25 MFI of untreated, non-co-cultured γδ cells. (E) Table shows the percentage increase in γδ CD25 expression of co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, non-co-cultured γδ cells. (F) Table shows the percentage increase in γδ CD25 expression for co-cultured cells treated with the highest concentration of vδ1 mAb compared to untreated, co-cultured and non-co-cultured γδ cells.

Cynomolgus monkey TCR down-regulation

Studies were performed to explore the ability of ADT1-4-2 to bind to cynomolgus monkey vδ1γδ T cells and induce their TCR down-regulation. In these studies cynomolgus PBMCs were collected from fresh blood and cultured for 14 days to expand γδ -T cells. Alternatively, prior to in vitro expansion, the αβ -T cells were depleted from the total PBMC population using magnetic beads coated with anti- αβ antibodies (clone R73). 250,000 cells were added to each well of a U-bottom 96-well plate in RPMI-1640 medium containing 10% FCS, antibiotics (P/S) and cytokine mixtures (IFNγ, IL21, IL4, IL 1. Beta. And IL 15) as follows. Seven days later, 10 μl of fresh medium containing 10% FCS, antibiotics (P/S) and cytokines IL21 and IL15 was added to the cells. 100 μl of medium was then replaced on day 11 with fresh medium containing 10% FCS, antibiotics (P/S) and IL 15. On day 14, cells were collected and TCR down-regulation assays were performed. The assay consisted of mixing cells with different concentrations of antibody for 2 hours. After incubation time, cells were stained for flow cytometry analysis of the following markers: CD3, TCR-. Alpha.beta., TCR-. Gamma.delta.and VD1. Live/dead cell dyes are also included to distinguish between live cell populations. Flow cytometry analysis was performed by gating cd3+, TCR- γδ+ and vd1+ cell populations, and VD1 mean fluorescence intensity was measured within the vd1+ gate. Only the highest concentration of isotype control (anti-RSV) or no antibody was used as negative control. The data were normalized to the control without any antibody. FIG. 38 (A) shows a comparison of the ability of ADT1-4-2 and ADT1-4 to bind to VD1 on cynomolgus gamma delta T cells and reduce VD1 expression on cynomolgus gamma delta T cells. The results showed that, following treatment with ADT1-4-2 (up to 76% at 33.3 nM), reduced dose response occurred in VD1 expression, whereas non-ADT 1-4 antibodies did not. In this particular assay, γδ -T cells are expanded from a population depleted of αβ -T cells. A 500-fold concentration range was used. FIG. 38 (B) shows the percentage of cell surface expression of VD1 after treatment with ADT 1-4-2. In this assay, γδ -T cells are expanded from all PBMC populations. A 10,000-fold concentration range was used. The data show reduced dose response of VD1 cell expression in 2 different donors (AD 8170 and AM945 BA) as an example. A total of 4 donors were tested and EC50 were extracted alone using GRAPHPAD PRISM nonlinear fit curve functions. Figure 38 (C) shows individual EC50 values for different donors as well as mean and standard deviation. The average value was 3.83.+ -. 2.23nM.

Antibody:

CD3-AF700 (BD Bioscience, clone SP34-2, reference number 557917)

TCR-. Alpha.beta. -AF647 (BioLegend, clone R73, reference number 201116)

TCR-. Gamma.delta. -BV421 (BioLegend, clone B1, reference number 331218)

VD1-PE (eBioscience, clone TS8.2, reference number 12-5679-42)

Additional studies were performed using whole blood samples from cynomolgus monkeys (n=5) (see fig. 38 (D-G)). ADT1-4-2 (0.052 mg/ml-cynomolgus monkey donor 1-2, or 1 mg/ml-cynomolgus monkey donor 3-5) was used to stimulate these samples for 20 hours. An untreated whole blood sample of each animal was used as a control. Viable single cd45+ monocytes were analyzed by flow cytometry after stimulation. For the flow assays in FIGS. 38D-G, labeled assay antibodies used include: anti-CD 3, BV421, SP34-2 BD Biosciences (catalog No. 562877), anti-CD 45, V500, D058-1283 BD Biosciences (561489), anti-full GD PE-Cy 7B 1 BioLegend (catalog No. 331222). First the results in (D) show that the absolute number of total viable γδ+ cells remains unchanged after stimulation with ADT1-4-2 compared to untreated controls. Although the absolute number of γδ T cells remained consistent, (E) subsequently showed that surface expression of both CD3 (Ei) and pan γδ TCR (Eii) on γδ T cells was down-regulated after ADT1-4-2 stimulation, as measured by a decrease in Median Fluorescence Intensity (MFI), compared to the unstimulated control. Representative flowsheet from cynomolgus monkey donor 1 is then shown in (F), where γδ T cells move obliquely to the lower left corner of the figure as the surface expression of CD3 and pan γδ is down-regulated following ADT1-4-2 stimulation, while overall pan γδ+% remains similar under both conditions. Finally, (G) shows, in contrast, that there is no change in surface expression of CD45 (a molecule not involved in TCR signaling) on γδ T cells after stimulation with ADT 1-4-2. These are key data illustrating the pharmacological activity of cynomolgus monkey cross-reactive, affinity matured anti-vδ1 antibodies as described herein on γδ T cells from cynomolgus monkeys.

Cytotoxicity enhancement of affinity matured clones

Affinity maturation of ADT1-4 and ADT1-7vδ1 clones significantly enhanced the cytotoxic effect of vδ1γδT cells in an in vitro THP-1 killing assay.

THP-1 cell killing assay

V delta 1 mAb and anti-RSV IgG controls were diluted to 1. Mu.g/ml in PBS and serially diluted 1:10 in PBS before addition to 384: 384 Kong Chaocheng image assay plates (PERKIN ELMER). THP-1 cells (ATCC) cultured in RPMI, 10% FCS (Invitrogen) were stained with [ 0.5. Mu.M ] CELLTRACE CFSE live cell stain for 20 min. The expanded skin-derived V.delta.1γ.delta.T cells were isolated from the tissue culture flask and resuspended in basal growth medium and then mixed with THP-1 cell suspension at 1:1. The cell suspension was seeded into 384 well assay plates at a final cell seeding density of 1,000 THP-1 cells per well and 2,000 vδ1γδ T cells per well. The mAb was diluted to a concentration in the range of 200ng/ml to 0.2pg/ml in the final assay volume. THP-1 and V.delta.1. Gamma. Delta. T cells were cultured in the presence of V.delta.1 mAb at 37℃under 5% CO ₂ for 24 hours. To determine the number of viable THP-1 cells after 24 hours, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on size, morphology, texture and intensity of viable cell staining.

FIG. 39 shows that affinity matured V.delta.1 mAb exhibited enhanced potency in THP-1 high content cytotoxicity assays as compared to the parental clone. (A and B) the number of viable THP-1 cells [ effector cells: target cells ratio 2:1 ] was quantified after 24 hours of co-culture with γδT cells in the presence of Vδ1 mAb or control. Cell amounts were normalized to the percentage of gamma delta T cell free control (100%). OKT3 (anti-CD 3 antibody) and anti-RSV isotype controls were included as assay controls. Cell numbers were calculated on living cells using a high content confocal microscope. The panels represent the parental ADT1-4 (A) and ADT1-7 (B) clones. (C) Bars represent average EC50 of OKT3 control, ADT1-4 and ADT1-7 clones. Data are expressed as mean ± standard deviation of n=3 biological replicates. (D) Table summarizes the EC50 s plotted in (C). Data are expressed as mean, standard deviation, fold change and percent improvement compared to ADT1-4 and ADT1-7 parental clones.

Effect of V.delta.1 monoclonal antibodies on Antibody Dependent Cellular Cytotoxicity (ADCC)

Expanded skin-derived vδ1γδ T cells from three donors were isolated from tissue culture flasks and resuspended in basal growth medium and seeded in white 96-well plates at a final density of 20,000 cells per well. Raji cells were also seeded at 20,000 per well and used as positive control for antibody rituximab-induced ADCC targeting CD 20. mAb was diluted and added to each well at a final concentration in the range of 1 to 100 nM. Fcyriiia+ ADD Bioassay effector cells were thawed and 62,500 cells were added to each well, followed by incubation at 37 ℃ for 4.5 hours at 5% co ₂. The Bio-Glo reagent containing fluorescein was added to each well, followed by incubation for an additional 10 minutes, and luminescence was measured using a plate reader.

Figure 40 shows that vδ1-targeted mabs induce negligible antibody-dependent cellular cytotoxicity (ADCC). ADCC is measured using an alternative reporter assay, wherein downstream NFAT signaling is measured by inducing luciferase activity within fcyriiia+ effector cells. Effector cells were co-cultured with three γδ T cell donors (effector cells: target cell ratio 3) and different concentrations of monoclonal antibodies. CD20 ⁺ Raji cells and rituximab were included as positive controls. Bioluminescence was measured after 4.5 hours of co-culture. Data are expressed as mean ± SD of three biological replicates (three γδ T cell donors).

Conclusion: the data indicate that ADT1-4 parent and affinity matured derivative ADT1-4-2 do not induce ADCC, as demonstrated by the FcgammaRIIIa+ effector reporter cell line (where NFAT signaling induces luciferase activity). However, OKT3 appears to induce high levels of ADCC-specific signaling.

Effect of V.delta.1 monoclonal antibodies on Complement Dependent Cytotoxicity (CDC)

The expanded skin-derived vδ1γδ T cells were isolated from the tissue culture flask and resuspended in basal growth medium and seeded in white 96-well plates at a final density of 75,000 cells per well. Raji cells were also seeded at 75,000 per well and used as a positive control for CD 20-targeting antibody rituximab-induced CDC. mAb was diluted and added to each well at a final concentration in the range of 1 to 100 nM. Fresh or heat inactivated human serum was added to a final concentration of 40%. The heat inactivation was performed by warming the serum in a 56 ℃ water bath for 30 minutes, followed by cooling on ice. The cultures were then incubated at 37℃for 24 hours with 5% CO ₂. To determine the number of living cells after 24 hours, cells were harvested and stained with xx viability dye at a dilution of 1:1000 for 20 minutes. Fluorescence was measured using MACSQuant and living cell counts were quantified based on the negative staining of the reactive dye.

The results are shown in fig. 41, which demonstrates that the vδ1-targeted mAb does not induce Complement Dependent Cytotoxicity (CDC). Number of γδ T cells surviving incubation with 40% human serum and 50nM anti-CD 3 OKT3 or anti-vδ1adt1-4-2 and different concentrations of monoclonal antibody. Serum was used fresh or heat inactivated beforehand. CD20 ⁺ Raji cells and rituximab were included as positive controls. Viability was measured by viability dye after 24 hours co-cultivation. Data are expressed as mean ± SD of three biological replicates.

Conclusion: the data indicate that ADT1-4-2 does not induce Complement Dependent Cytotoxicity (CDC) and that cytotoxicity of V.delta.1 positive cells does not increase in the presence of complement containing serum. Rituximab, however, exhibits a strong CDC-stimulating effect on CD20 positive Raji cells.

Healthy cell retention

Studies were conducted to explore the effect of stimulation/activation of vδ1 cells with anti-vδ1 antibodies on cytotoxicity against healthy cells. This was tested by incubating anti-vδ1 clone ADT1-4-2 with PBMCs and then assessing cytotoxicity and apoptosis of monocytes.

Cryopreserved human Peripheral Blood Mononuclear Cells (PBMCs) were commercially derived from 3 healthy donors. PBMC were seeded into round bottom 96 well tissue culture plates at 250,000 cells/well in 250ul of complete medium (RPMI supplemented with 10% FCS, penicillin/streptomycin, non-essential amino acids, sodium pyruvate and HEPES) with 10ng/ml IL 15. Titration of vδ1 antibody ADT1-4-2 to a final concentration of 6.6nM was added. RSV IgG, igG2 alpha and OKT3 antibodies were included as controls. Stimulation was performed for 20 hours. Flow cytometry analysis was performed at the endpoint to determine monocyte phenotype and apoptotic monocytes under various conditions. Cells were first gated on live single cells, followed by CD14 (Miltenyi 130-110-523) to identify monocytes. Apoptotic monocytes were then identified by positive ApoTracker Green staining (Biolegend 427402).

Fig. 42A (upper left panel) shows vδ1TCR stained MFI as an indication of target engagement following antibody stimulation. Fig. 42B (upper right panel) and xixC (lower panel) show the end-point monocyte phenotype and apoptotic monocytes (respectively) after antibody stimulation. Stimulation of vδ1 antibody clone ADT1-4-2 was applied to vδ1 cells, but did not cause cytotoxicity of healthy monocytes.

Example 25 Effect of anti-V.delta.1 antibodies on TIL populations from primary tumor biopsies

Anti-vδ1 antibodies promote activation, proliferation and cytotoxicity of tumor-infiltrating lymphocytes.

Studies were performed to explore the modulation and proliferation of human Tumor Infiltrating Lymphocytes (TILs) conferred by anti-vδ1 antibodies. For these studies, human Renal Cell Carcinoma (RCC) tumor biopsies were freshly delivered and treated after receipt. Biopsies were cut into small pieces measuring about 2mm ² and TIL was obtained using a modification of the method described initially by Kupper and Clarke (Clarke et al, 2006). Specifically, up to four 2mm ² biopsies were placed on a 9mm x 1.5mm Cellfoam matrix and one matrix was placed on each well of a 24 well plate. The biopsies were then cultured in 2ml of Iscove's Modified Dulbecco's Medium (IMDM) supplemented with 4% human plasma, beta-mercaptoethanol (50. Mu.M), penicillin (100U/ml), streptomycin (100. Mu.g/ml), amphotericin B (2.5. Mu.g/ml), HEPES (10 mM), sodium pyruvate (1 mM), MEM nonessential amino acid solution (1X) and IL-15 (2 ng/ml, miltenyi Biotech). 1ml of medium was aspirated every 3-4 days and replaced with 1ml of complete medium containing 2-fold concentrated IL-15. TIL was harvested 10 or 11 days later, centrifuged at 300×g for 5min through a 70 μm nylon cell filter, and resuspended in complete medium for counting. 400,000 cells were then seeded into 96-well plates and then stimulated with anti-vδ1 antibodies. TIL was stimulated with ADT1-4, ADT1-4-2, ADT1-7-3 or RSV IgG1 isotype control antibodies in the presence of IL-15 at a concentration of 2 or 50 ng/ml.

Fig. 43 (a and B) shows a decrease in vδ1 TCR expression on total tumor infiltrating- γδ T cells after mAb stimulation 48 (a) or 72 (B) hours in two independent donors, confirming target engagement in each case. Tumor infiltrating vδ1 ⁺ cells were analyzed for CD25 and Ki67 expression. FIG. 43 (C) shows enhanced expression of CD25 and Ki67 on V.delta.1 ⁺ T cells after 48 hours of stimulation with ADT1-4-2 compared to IgG1 isotype control or ADT1-4 stimulation. mAb stimulated production of cytokines in TIL culture supernatants was analyzed by MSD. FIG. 43 (D) shows a significant fold increase in IFN-. Gamma.production by TIL stimulated with ADT1-4-2 or ADT1-7-3 in the presence of 50ng/ml IL-15 for 72 hours. FIG. 43 (E) shows that stimulation of TIL with ADT-1-4-2 or ADT1-7-3 did not enhance secretion of type 17 associated cytokines IL-6 or IL-17 at this time.

Or biopsies from different donors are subjected to enzymatic digestion after receipt to obtain single cell suspensions. Specifically, up to 1g of tissue was placed in MILTENYI C tubes along with 4.7mL of RPMI supplemented with enzyme from Miltenyi tumor dissociation kit at the manufacturer's recommended concentration, which was used at 0.2x concentration to prevent the associated cell surface molecules from lysing, except for enzyme R. The C-tube was placed on GENTLEMACS ^TM Octo ionizer with a heating block attached. Program 37c_h_tdk_1 was selected for dissociation of small tumors. After 1 hour, the digests were filtered through a 70 μm filter and full IMDM containing 4% human plasma was added to quench the enzymatic activity. Cells were then washed twice with full IMDM and resuspended for counting. Depending on the number of cells, 2X 10 ⁶ or 4X 10 ⁶ cells per well were plated in 48-well plates and stimulated with ADT1-4, ADT1-4-2 or RSV IgG1 isotype control antibody in the presence of IL-15 at a concentration of 2 ng/ml. The TIL isolated by enzymatic digestion was analyzed by flow cytometry 24-72 hours after mAb stimulation. Fig. 43 (F and G) shows the decrease in vδ1 TCR expression on total tumor infiltrating γδ T cells 24 (F) or 72 (G) hours after mAb stimulation in two individual donors confirming target engagement in TIL isolated by enzymatic digestion. FIG. 43 (H) shows a dose-dependent increase in Ki67 expression on γδ T cells after 72 hours of stimulation with ADT 1-4-2. mAb stimulated production of cytokines in TIL culture supernatants was analyzed by MSD. FIG. 43 (I and J) shows fold increases in IFN-gamma production by TIL isolated from two individual donors by enzymatic digestion at two time points for 24 hours (I) and 72 hours (J) and stimulated with ADT1-4-2 at a concentration of 6.66nM in the presence of 2ng/ml IL-15. Taken together, these combined results highlight the ability of affinity matured anti-vδ1 antibodies to drive activation, proliferation and anti-tumor cytokine production of tumor infiltrating γδ T cells within 72 hours of stimulation.

In another experiment, TILs derived from lung tumors were isolated on a grid matrix and stimulated with ADT1-4, ADT1-4-2 or IgG1 isotype control in the presence of 2ng/ml IL-15 for 10 days. FIG. 44 (A) shows enhanced expression of CD25 and Ki67 in γδ T cells stimulated with ADT1-4-2 for 10 days. FIG. 44 (B) shows that perforin+granzyme B ⁺ γδ T cells were greatly increased after 10 days of stimulation with ADT 1-4-2. Longer stimulation with mAb allowed analysis of the immune permissivity of αβ T cells by vδ1 ⁺ T cells stimulated with anti-vδ1 antibody. FIG. 44 (C) shows that granzyme B and perforin expression of CD8 ⁺ and CD8 ^-. Alpha. Beta. T cells was significantly enhanced 10 days after tumor infiltration of V.delta.1 ⁺ T cells stimulated with ADT 1-4-2. Supernatants of TIL incubated with mAb for 10 days were analyzed by MSD. FIG. 44 (D) shows a significant increase in IFN-gamma production of lung tumor-derived TIL and a modest increase in IL-17 and IL-6 production following stimulation with ADT 1-4-2. FIG. 44 (E) demonstrates enhancement of the production of the chemokines CCL2, CCL4 and CXCL10 by TIL after 10 days of stimulation with ADT 1-4-2. Taken together, these results underscore that anti-vδ1 antibodies can drive activation and cytotoxicity in tumor-infiltrating γδt cells, which in turn can promote cytotoxicity in tumor-infiltrating αβt cells.

EXAMPLE 26 binding affinity of V.delta.1-CD 19 bispecific antibodies to human and cynomolgus monkey V.delta.1

Binding affinity of the antibodies to the target (i.e., the vδ1 chain of γδ TCR, both human and cynomolgus monkey antigens) was established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). The antibody was coated onto Planar protein a sensor chip (Reichert Technologies) to increase baseline by about 500: 500 uRIU. Recombinant human vδ1 heterodimer or cynomolgus vδ1 heterodimer flows through the cells. All experiments were performed at room temperature. The results are shown in fig. 49.

These data indicate that the bispecific antibody binds both human and cynomolgus monkey vδ1.

Example 27A V.delta.1 gamma.delta.T cell activation and V.delta.1-CD 19 in cytotoxicity assays

Bispecific antibodies

Healthy B cells were isolated from PBMC (Lonza) using negative magnetic activated cell sorting (MACS; miltenyiBiotec). The expression of CD19 on cancerous NALM-6 cells (ATCC), raji cells (ATCC) and isolated healthy B cells was determined. Briefly, 5×10≡4 cells were incubated with CD19 antibodies (Biolegend) for 15min at 4℃and then washed and fixed. CD19 expression was determined by flow cytometry (MACSQuant, miltenyiBiotec). The results are shown in fig. 50A.

The effect of CD19-vδ1 bispecific antibodies on CD19 ⁺ target cytotoxicity and CD19 ⁺ healthy cell retention was determined using high content confocal imaging in Opera Phenix (PERKIN ELMER). NALM-6, raji and B cells were stained with [ 0.5. Mu.M ] CELLTRACE CFSE live cell stain for 20 min. Bispecific antibodies and controls were serially diluted in PBS and then added to 384-well imaging assay plates (PERKIN ELMER). The expanded skin-derived V.delta.1γ.delta.T cells were isolated from the tissue culture flask and resuspended in basal growth medium and then mixed in 1:1 suspension with NALM-6, raji or B cells. The cell suspension was seeded into 384 well assay plates to give a final cell seeding density of 2,000 NALM-6, raji or B cells per well and 2,000 V.delta.1γ.delta.T cells per well. The final antibody concentration was determined to be in the range of 6.6nM to 66 pM. Cells were co-cultured for 24 hours and then stained with DRAQ7 (final 1:300, abcam). To determine the number of live target cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on the size, morphology, texture and intensity of CFSE staining and absence of DRAQ7 staining. The results are shown in fig. 50.

To determine the effect of CD19-vδ1 bispecific antibodies on vδ1γδt cell activation and degranulation, vδ1 TCR downregulation and CD107a upregulation were quantified in the presence of CD19 ⁺ target cells and CD19 ⁺ healthy cells. Briefly, antibodies were serially diluted in PBS and then added to a U-bottom 96-well plate. NALM-6 and isolated B cells were stained with [ 0.5. Mu.M ] CELLTRACE CFSE live cell stain for 20 minutes. The skin-derived V.delta.1γ.delta.T cells were isolated from the flask and the cell suspension was mixed with NALM-6 or B cells at 0.5:1. The cell suspension was seeded into assay plates at 2.5X10-4 V.delta.1γ.delta.T cells per well to 5X 10-4 NALM-6 or B cells per well. The final antibody concentration was determined to be in the range of 60nM to 3 pM. Cells were incubated at 37℃for 4 hours at 5% CO ₂. Cells were washed and stained for surface expression for 30min at 4 ℃ for dead cells (eFlour, invitrogen), vδ TCR (MiltenyiBiotec) and aCD107a (Miltenyi). Cells were washed in FACS buffer and resuspended in Cell Fix (BD sciences) and then incubated overnight at 4 ℃ in the dark. The following day VD1 TCR expression levels were measured by flow cytometry using MACS Quant Analyzer. The results are shown in fig. 50F to K.

Conclusion: the vδ1-CD19 bispecific antibody enhances γδ T cell mediated cytotoxicity of CD19 ⁺ target cells while retaining healthy CD19 ⁺ cells. Affinity maturation of vδ1 antibodies enhanced cytotoxicity compared to the monovalent format of the parental clone.

Example 27B: the vδ1-CD19 bispecific antibody selectively enhances the cytotoxic effect of vδ1γδ T cells on cancer cells while retaining healthy cells

The effect of CD19-vδ1 bispecific antibodies on cd19+ target cytotoxicity and cd19+ healthy cell retention was determined using high content confocal imaging in Opera phenoix (PERKIN ELMER). Healthy B cells were isolated from PBMC (Lonza) using negative magnetic activated cell sorting (MACS; miltenyiBiotec). Raji cells and healthy primary B cells were stained with CELLTRACE dye. The expanded skin-derived vδ1γδ T cells and donor-matched αβ T cells were removed from the culture, washed and resuspended in medium, and then mixed 1:1 with Raji cells and B cells. Bispecific antibodies and controls were serially diluted in PBS and then added to 384-well imaging assay plates (PERKIN ELMER). Cell suspensions of γδ T cells, raji cells and B cells, or αβ T cells, raji cells and B cells, or vδ1γδ T cells, αβ T cells, raji cells and B cells were added to 384 well imaging assay plates to achieve 2,000 per well of each cell type, and the final antibody assay concentration was in the range between 6.6nM to 66 pM. To determine the number of live target cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on size, morphology, texture and intensity of CFSE staining at 24 hours. The results are shown in fig. 63A to F.

To determine if activation of vδ1γδ T cells or αβ T cells resulted in increased production of the pro-tumor cytokine IL-17, supernatants collected from imaging plates after 24 hours of imaging were obtained. Supernatants were run on U-Plex 10-Plex Meso Scale Discovery (MSD) to quantify IL-17A levels. The results are shown in fig. 63G to I.

IL-17A (interleukin-17A) is a pro-tumor cytokine produced by activated T cells. IL-17A can enhance tumor growth and inhibit anti-cancer immune responses. As shown in fig. 63G to I, the anti-vδ1 antibody did not induce secretion of IL-17A, whereas the anti-CD 3 antibody induced secretion of IL-17A.

Conclusion: the vδ1-CD19 bispecific antibody enhances γδ T cell mediated cytotoxicity of cd19+ target cells while retaining healthy cd19+ cells. In contrast, anti-CD 3 xcd 19 bispecific antibodies enhance γδ T cell and αβ T cell mediated lysis of cd19+ target cells, but activation of αβ T cells also enhances lysis of healthy primary cd19+ B cells, as well as secretion of the tumorigenic cytokine IL-17A.

EXAMPLE 28 V.delta.1-Her 2 bispecific antibodies in binding and high content cytotoxicity assays

Expression of Her2 and V.delta.1 was determined on SK-BR-3 cells (Caltag-MEDSYSTEMS LTD), BT-474 cells (ATCC), MDA-MB-231-Luc cells (Creative Biogene Biotechnology) and V.delta.1. Gamma. Delta.T cells. Briefly, 5×10≡4 cells were incubated with Her2 and vδ1 antibodies (MiltenyiBiotec) for 15 min at 4 ℃ before washing and fixing. Expression of Her2 and vδ1 was determined by flow cytometry (MACSQuant, miltenyiBiotec). The results are shown in fig. 51A and 51B. Binding of Her2-vδ1 bispecific antibody was determined by incubating target cells with a series of concentrations of ADT1-4 or ADT1-4-2 mAb or vδ1 bispecific antibody or control (IgG control or trastuzumab) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 51C to F.

The effect of Her2-vδ1 bispecific antibodies on Her2+/Her 2-target cytotoxicity was determined using high content confocal imaging in Opera Phenix (PERKIN ELMER). Briefly, SK-BR-3, BT-474 and MDA-MB-231 cells were seeded into 384 well imaging plates (PERKIN ELMER) to give a final seeding density of 2,000 target cells per well, and then incubated overnight at 37℃at 5% CO 2. The antibodies were diluted in PBS and serially diluted 1:10 and then added to the assay plate to give final assay concentrations of 333nM to 0.33 pM. The expanded skin-derived vδ1γδ T cells were isolated from the tissue culture flask and resuspended in basal growth medium, then effector cells at 1:1: target cell ratio was added to the assay plate at 2,000 cells per well. Cells were co-cultured for 24 hours and then stained with Hoechst (final 1:1000, invitrogen) and DRAQ7 (final 1:300, abcam). To determine the number of live target cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on the size, morphology, texture and intensity of viable cell staining, and absence of DRAQ7 staining. The results are shown in fig. 51G to J.

Conclusion: the vδ1-Her2 bispecific antibody enhances γδ T cell mediated cytotoxicity of Her2 ⁺ target cells while retaining healthy Her 2-cells. Affinity maturation of the V.delta.1 antibody (ADT 1-4-2) enhanced cytotoxicity compared to the parental clone (ADT 4-1).

EXAMPLE 29 binding affinity of V.delta.1-EGFR bispecific antibodies to human V.delta.1 and human EGFR antigens

Binding affinity of the antibodies to the target (i.e., vδ1 chain of γδ TCR and EGFR) was established by SPR analysis using Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500 uRIU. Recombinant human vδ1 heterodimer or human EGFR was flowed through cells at a concentration of 100nM with the following parameters: 180 seconds association, 480 seconds dissociation, flow rate 25. Mu.L/min, running buffer PBS+0.05% Tween 20. All experiments were performed at room temperature. The results are shown in fig. 52A and 52B.

Conclusion: this data shows that despite the introduction of human EGFR binding capacity, the vδ1/EGFR bispecific antibody also shows binding comparable to human vδ1 and monospecific anti-vδ1 antibodies used in its preparation.

EXAMPLE 30 target cell binding of V.delta.1-EGFR bispecific antibody

EGFR positive A431 and V.delta.1 positive primary gamma.delta.T cells were assayed to determine the specificity and affinity of EGFR/V.delta.1 bispecific antibody binding. Target cells were isolated from tissue culture flasks, resuspended in PBS, and seeded in v-bottom 96-well plates at a final density of 100,000 cells per well. The cells were centrifuged and the cell pellet resuspended in FcR blocking reagent according to the manufacturer's instructions and incubated for 20 minutes at 4 ℃ before further washing. Antibodies were diluted to 500nM in PBS and serially diluted 1:10 to 50pM in PBS and added to cells, followed by incubation at 4℃for 20 minutes. To determine the number of mAbs bound to the cell surface, cells were stained with murine anti-human IgG secondary antibodies conjugated to APC (dilution: 1:100), except for the vital dye. After incubation at 4 ℃ for 20 minutes, the cells were washed twice and fluorescence was measured using MACSQuant. The results are shown in fig. 53.

Conclusion: this data shows that, despite the introduction of binding to the EGFR positive a431 cell line, the vδ1/EGFR bispecific antibody also shows binding to vδ1γδ T cells comparable to the monospecific anti-vδ1 antibody used in its preparation.

EXAMPLE 31 in vitro assessment of γδ T cell activation and target cytotoxicity

Expanded skin-derived vδ1γδ T cells and a431 cells were isolated from the tissue culture flask and resuspended in basal growth medium and, depending on the desired effector cells: target cell ratios were seeded in 96-well plates at relevant cell dilutions. mAb was diluted and added to each well at the indicated concentration. The cultures were then incubated at 37℃under 5% CO2 for 4 (D) or 24 hours (A-C, E). To determine the number of living cells, the cells were harvested and stained with a vital stain at a dilution of 1:1000 for 20 minutes. To determine CD25 status, cells were surface stained with anti-CD 25 antibodies after cell harvest. To measure degranulation, fluorophore conjugated anti-CD 107 a antibodies were added directly to the cell antibody mixture at the beginning of co-culture. Subsequently, two washes and cell fixation were performed, fluorescence was measured using MACSQuant, and viable cell count and median fluorescence intensity were determined. The results are shown in fig. 54.

Conclusion: the data indicate that the vδ1/EGFR bispecific antibody induces activation and degranulation of primary vδ1-positive γδ T cells, resulting in an increased cell-mediated lysis of the EGFR-positive a431 cell line.

Example 32 further evaluation of the γδ T cell activation and non-depletion effects conferred by the antibodies of the invention

Study of blood-derived vδ1+ cells (fig. 55A, B):

studies were performed to explore the effect of stimulation/activation of vδ1 cells with anti-vδ1 antibodies on CD3 down-regulation on vδ1 cells. This was done by incubating anti-vδ1 clone ADT1-4-2 with PBMCs and then analyzing the TCR test by phenotypic analysis.

Cryopreserved human Peripheral Blood Mononuclear Cells (PBMCs) were of commercial origin and inoculated into round bottom 96-well tissue culture plates at 250,000 cells/well in 250ul complete medium (RPMI supplemented with 10% fcs, penicillin/streptomycin, nonessential amino acids, sodium pyruvate, and HEPES) with 10ng/ml IL 15. The v.delta.1 antibody ADT1-4-2 was titrated to a final concentration of 1ug/ml (6.67 nM), 0.01ug/ml (0.067 nM), or 0.0001ug/ml (0.00067 nM). RSV IgG antibodies were included as controls for matched concentration. Cultures were incubated for 14 days, with medium and antibody supplementation every 3 days. Flow cytometry analysis was performed at the endpoint to phenotype vδ1 cells and TCR expression under various conditions. Cells were first gated on living single cells, followed by ubiquity δ (MILTENYI REA592; 130-113-508), which is the parent gate of vδ1 (MILTENYI REA173; 130-100-553), and vδ1 itself is the parent gate of CD3 (MILTENYI REA613; 130-113-142). The cell population was identified by positive staining, and then the relative expression level of each marker between samples was identified by MFI.

Study of tumor resident v61+ cells (fig. 55C, D, E):

For fig. 55C, D and E, solid tumor biopsies (resected from Renal Cell Carcinoma (RCC) patients) are freshly delivered and processed after receipt to obtain single cell suspensions. Specifically, tissues were cut into small pieces measuring about 2mm2 and up to 1g of tissue was placed in MILTENYI C tubes along with 4.7mlRPMI, RPMI was supplemented with enzyme from Miltenyi tumor dissociation kit at the manufacturer recommended concentration, which was used at 0.2X concentration except for enzyme R to prevent lysis of the relevant cell surface molecules. The C-tube was placed on GENTLEMACSTM OCTO ionizer with a heating block attached. The procedure 37c_h_tdk_1 was selected for dissociating soft tumors. After 1 hour, the digests were filtered through a 70 μm filter and full IMDM containing 4% human plasma was added to quench the enzymatic activity. Cells were then washed twice with full IMDM and resuspended for counting. At this time, the cells were spread for stimulation. 2.5X106 cells per well were plated in 24-well plates and stimulated with ADT1-4-2 or anti-IgG 1 isotype control antibody in the presence of IL-15 at a concentration of 2 ng/ml. Viable single cd45+ Tumor Infiltrating Lymphocytes (TILs) were analyzed by flow cytometry 24 hours after mAb stimulation.

Fig. 55A shows vδ1 TCR MFI as an indication of mAb target engagement following antibody stimulation. Fig. 55B shows MFI for CD3 expression on positively gated vδ1 cells. Stimulation of ADT1-4-2 using the vδ1 antibody clones engaged vδ1 cells, resulting in down-regulation of vδ1 and CD3 on vδ1 cells. Combined figures 55C, D and E show that activating tumor-associated vδ1T cells with an exemplary anti-vδ1 antibody as described herein does not deplete vδ1+ T cells, but does bind and activate the CD3/γδ TCR complex and results in its down-regulation and loss of surface expression. Specifically, FIG. 55C shows that the absolute numbers of total γδ+ and γδ+Vδ1+ cells per well remained constant after stimulation with ADT-1-4-2 or anti-IgG 1 isotype control. Although the absolute numbers of vδ1T cells remained consistent, figure 55D shows that surface expression of vδ1 TCR, pan γδtcr and CD3 was down-regulated on vδ1T cells after stimulation with ADT-1-4-2, but not in the case of anti-IgG 1 isotype control. In contrast, the surface expression of CD45, a molecule not involved in TCR signaling, did not change after ADT-1-4-2 stimulation. FIG. 55E shows a FACS diagram (mAb concentration 33.3 nM) representing the data of FIG. 55D, in which V.delta.1T cells are marked gray. Here, vδ1+ T cells were moved obliquely to the lower left corner of the figure, because surface expression of CD3 and pan γδ was down-regulated after stimulation with ADT-1-4-2, but not down-regulated in the case of isotype control (as shown in the upper panel), but overall pan γδ+% in cd45+ remained similar in both cases. Further gating of vδ1 expression (lower panel) specifically shows that vδ1 MFI decreases after stimulation, so that a small portion (about 5%) moves from the established vδ1 TCR positive gate to the vδ1 'negative' gate (about 5% increase is observed with the 'negative' gate). In combination, these are key data demonstrating that activation of vδ1 TCR with an antibody as described herein does not deplete vδ1+ T cells. From these findings, etc., it is contemplated that the antibodies of the invention may be used as a medicament for treating a disease or disorder, thereby ameliorating at least one sign or symptom of the disease or disorder by a mechanism involving activation of blood, tissue, and tumor-resident vδ1+ T cells.

Example 33: vδ1-fapα bispecific antibodies enhance vδ1γδ T cell activation and fapα+ fibroblast lysis

Binding kinetics for binding of anti-vδ1 (ADT 1-4-2), anti-fapα (based on sibutrab) and anti-vδ1×fapα (ADT 1-4×sibutrab) (SEQ ID NO:401 or SEQ ID NO:414 and SEQ ID NO: 415) antibodies to their targets (i.e. vδ1 chain of γδtcr and fapα) were established by SPR analysis using Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500 uRIU. Recombinant human vδ1 heterodimer or human fapα flowed through cells at a concentration of 100nM with the following parameters: 180 seconds association, 480 seconds dissociation, flow rate 25. Mu.L/min, running buffer PBS+0.05% Tween 20. All experiments were performed at room temperature. The results are shown in fig. 56A. The KD values are as follows:

Binding of vδ1-fapα bispecific antibodies was determined by incubating fapα ⁺ target cells or vδ1 ⁺ effector cells with a range of concentrations of anti-vδ1 antibodies or vδ1 bispecific antibodies or controls (IgG control or anti-fapα) for 15 minutes. After washing, cells were incubated with anti-human Fc secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 56B to C.

To determine the effect of vδ1-fapα bispecific antibodies on vδ1γδ T cell activation and degranulation, vδ1 TCR down-regulation and CD107a up-regulation were quantified in the presence of fapα ⁺ target cells. Briefly, anti-vδ1, anti-fapα and anti-vδ1×fapα bispecific antibodies were serially diluted in PBS and then added to a U-bottom 96-well plate. FAP alpha ⁺ target cells (BJ fibroblasts or human dermal fibroblasts) were stained with [ 0.5. Mu.M ] CELLTRACE CFSE live cell dye for 20 min. The skin-derived V.delta.1γ.delta.T cells were isolated from the flask and the cell suspension was mixed 1:1 with FAP.alpha. ⁺ target cells or diluted 1:1 with medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4 FAP.alpha. ⁺ target cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃, 5% co ₂ for 4 hours, then washed and surface-expression stained for dead cells (eFlour, invitrogen), vδ1 TCR (MiltenyiBiotec) and aCD107a (Miltenyi) at 4 ℃ for 30 minutes. Cells were washed in FACS buffer and resuspended in Cell Fix (BD sciences) and then incubated overnight in the dark at 4 ℃. The following day VD1 TCR and CD107a expression levels, as determined by Median Fluorescence Intensity (MFI), were measured by flow cytometry using MACS Quant Analyzer. The results are shown in FIGS. 56D-G.

Moderate vδ TCR down-regulation was observed in the absence of fapα ⁺ fibroblasts (fig. 56, d). However, in the presence of fapα+ fibroblasts, anti-fapα -vδ1 bispecific antibodies were greatly enhanced, TCR down-regulated (fig. 56, e). This demonstrates that the effect of anti-fapα -vδ1 bispecific antibodies on vδ1 TCR down-regulation on vδ1γδ T cells is enhanced by binding to tumor-specific antigens, such as fapα, on neighboring cells.

In the presence of anti-vδ1xfapα bispecific antibody and fapα ⁺ fibroblasts, CD107a (degranulation marker) was upregulated on vδ1γδ T cells compared to the monoclonal control (fig. 56, g), while in the absence of fapα ⁺ fibroblasts, CD107a was not upregulated on vδ1γδ T cells compared to the monoclonal control (fig. 56, f), demonstrating that the effect of anti-fapα -vδ1 bispecific antibody on CD107a upregulation on vδ1γδ T cells is specific, and anti-fapα -vδ1 bispecific antibody enhanced degranulation and activation of vδ1γδ T cells in the presence of fapα ⁺ cells.

The effect of fapα -vδ1 bispecific antibodies on fapα ⁺ target cytotoxicity was determined using high content confocal imaging in Opera Phenix (PERKIN ELMER). FAP alpha ⁺ target cells (BJ fibroblasts, human dermal fibroblasts) were seeded into 384 well imaging plates (PERKIN ELMER) and incubated for 24 hours. Bispecific antibodies and controls were serially diluted in PBS and then added to the assay plates. Amplified skin-derived V.delta.1γ.delta.T cells were isolated from tissue culture flasks and resuspended in basal growth medium and then added to assay plates at a ratio of effector to target of 1:1. The final measured antibody concentration was in the range between 6.6nM and 66 fM. Cells were co-cultured for 24 hours and then stained with DRAQ7 (final 1:300, abcam) and Hoechst (final 1:1000, invitrogen). To determine the number of live target cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on the size, morphology, texture and intensity of Hoechst staining and absence of DRAQ7 staining. The results are shown in fig. 56, h. In the presence of anti-fapα -vδ1 bispecific antibodies, a significant increase in fibroblast cytotoxicity was observed compared to anti-vδ1 and anti-fapα controls (fig. 56, h). This suggests that direct bridging of vδ1γδ T cells with target cells can specifically enhance the activation and cytotoxic effects of vδ1γδ T cells.

Conclusion: the anti-vδ1-fapα bispecific antibody specifically binds vδ1 ⁺ γδ T cells and fapα+ target cells, thereby enhancing activation of vδ1γδ T cells in the presence of fapα ⁺ cells, which is manifested by elevated vδ1 TCR down-regulation, CD107a up-regulation, and fapα ⁺ fibroblast lysis.

Example 34: vδ1-MSLN bispecific antibodies enhance vδ1γδ T cell activation and MSLN ⁺ target cell lysis

Binding kinetics for binding of anti-V.delta.1 (ADT 1-4-2), anti-MSLN (mesothelin) (based on the antibodies disclosed in US 2014/0004121) and anti-V.delta.1 XMSLN (ADT 1-4-2 XMSLN) (SEQ ID NO:403 or SEQ ID NO:414 and SEQ ID NO: 416) antibodies to their targets (i.e., V.delta.1 chains of γδ TCR and MSLN) were established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human vδ1 heterodimer or human MSLN flowed through cells at a maximum concentration of 100nM with the following parameters: 180 seconds association, 480 seconds dissociation, flow rate 25. Mu.L/min, running buffer PBS+0.05% Tween 20. All experiments were performed at room temperature. The results are shown in fig. 57A. The KD values are as follows:

Binding of MSLN-vδ1 bispecific antibody was determined by incubating MSLN ⁺ target cells or vδ1 ⁺ effector cells with a range of concentrations of anti-vδ1 antibody or vδ1 bispecific antibody or control (IgG control or anti-MSLN) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 57B to C.

To determine the effect of MSLN-vδ1 bispecific antibodies on vδ1γδt cell activation and degranulation, vδ TCR down-regulation and CD107a up-regulation were quantified in the presence of MSLN ⁺ target cells. Briefly, anti-vδ1, anti-MSLN, and anti-vδ1×msln bispecific antibodies were serially diluted in PBS and then added to U-bottom 96-well plates. MSLN ⁺ target cells (OVCAR-3 or HeLa) were stained with [ 0.5. Mu.M ] CELLTRACE CFSE live cell stain for 20 min. The skin-derived V.delta.1γ.delta.T cells were isolated from the flask and the cell suspension was mixed 1:1 with MSLN ⁺ target cells or diluted 1:1 with medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4 MSLN ⁺ target cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃, 5% co ₂ for 4 hours, then washed and surface-expression stained for dead cells (eFlour, invitrogen), vδ1 TCR (MiltenyiBiotec) and aCD107a (Miltenyi) at 4 ℃ for 30 minutes. Cells were washed in FACS buffer and resuspended in Cell Fix (BD sciences) and then incubated overnight in the dark at 4 ℃. The following day VD1 TCR and CD107a expression levels, as determined by Median Fluorescence Intensity (MFI), were measured by flow cytometry using MACS Quant Analyzer. The results are shown in FIGS. 57D-G.

Moderate vδ TCR down-regulation was observed in the absence of MSLN ⁺ target cells (fig. 57, d). However, in the presence of MSLN ⁺ OVCAR-3 cells, the anti-MSLN-vδ1 bispecific antibody greatly enhanced TCR down-regulation (fig. 57, e). This demonstrates that the effect of anti-MSLN-vδ1 bispecific antibodies on vδ1 TCR down-regulation on vδ1γδ T cells is enhanced by binding to tumor specific antigens such as MSLN on neighboring cells.

In the presence of anti-vδ1×msln bispecific antibody and MSLN ⁺ OVCAR-3 cells, CD107a (degranulation marker) was upregulated on vδ1γδ T cells compared to the monoclonal control (fig. 57, g), while in the absence of MSLN ⁺ OVCAR-3 cells, CD107a was not upregulated on vδ1γδ T cells compared to the monoclonal control (fig. 57, f), demonstrating that the effect of anti-MSLN-vδ1 bispecific antibody on CD107a upregulation on vδ1γδ T cells is specific, and anti-MSLN-vδ1 bispecific antibody enhances degranulation and activation of vδ1γδ T cells in the presence of MSLN ⁺ cells.

The effect of MSLN-vδ1 bispecific antibody on MSLN ⁺ target cytotoxicity was determined using high content confocal imaging in Opera Phenix (PERKIN ELMER). MSLN ⁺ target cells (HeLa or OVCAR-2) were seeded into 384 well imaging plates (PERKIN ELMER) and incubated for 24 hours. Bispecific antibodies and controls were serially diluted in PBS and then added to the assay plates. The expanded skin-derived vδ1γδ T cells were isolated from the tissue culture flask and resuspended in basal growth medium, followed by effector cells at 1:1: target cell ratios were added to assay plates. The final antibody concentration was determined to be in the range between 6.6nM and 66 fM. Cells were co-cultured for 24 hours and then stained with DRAQ7 (final 1:300, abcam) and Hoechst (final 1:1000, invitrogen). To determine the number of live target cells, nine fields of view were captured at 10 x magnification using an Opera Phenix high content platform to obtain confocal images. Viable cell counts were quantified based on the size, morphology, texture and intensity of Hoechst staining and absence of DRAQ7 staining. The results are shown in fig. 57H. In the presence of anti-MSLN-vδ1 bispecific antibodies, a significant increase in target cytotoxicity was observed compared to anti-vδ1 and anti-MSLN controls (fig. 57, h). This suggests that direct bridging of vδ1γδ T cells with target cells can specifically enhance the activation and cytotoxic effects of vδ1γδ T cells.

Conclusion: anti-vδ1×msln bispecific antibodies specifically bind vδ1 ⁺ γδ T cells and MSLN ⁺ target cells, thereby enhancing activation of vδ1γδ T cells in the presence of MSLN ⁺ cells, which is manifested by elevated vδ1 TCR down-regulation, CD107a up-regulation, and lysis of MSLN ⁺ cells.

Example 35: vδ1-PD-1 bispecific antibodies enhance vδ1γδ T cell activation and block PD-1/PD-L1 checkpoint inhibition

Binding kinetics for binding of anti-V.delta.1 (ADT 1-4-2), anti-PD-1 (based on palbociclizumab) and anti-V.delta.1X PD-1 (ADT 1-4-2X palbociclizumab) (SEQ ID NO:405 or SEQ ID NO:438 and SEQ ID NO: 417) antibodies to the targets (i.e., V.delta.1 chain of γδ TCR and PD-1) were established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human V.delta.1 heterodimer or human PD-1 flowed through cells at the highest concentration of 100 nM. The results are shown in fig. 58A. The KD values are as follows:

To assess dual binding of the bispecific antibody to both target ligands, recombinant PD-1 was first immobilized on a carboxymethyl dextran sensor chip (Reichert Technologies) at 10ug/ml and then flowed through the bispecific antibody at 100 nM. The ability to subsequently bind vδ1γδtcr was then assessed by flowing 100nM through recombinant human vδ1 heterodimer. All experiments were performed at room temperature. The results are shown in fig. 58B.

Binding of anti-vδ1×pd-1 bispecific antibodies to vδ1γδ T cells and PD-1 ⁺ immune cells was assessed by flow cytometry. Initially, CD4 and CD 8T cells were negatively selected by magnetic sorting through PBMC buffy coats extracted from whole blood. Cell surface expression of PD-1 was detected on CD4 and CD 8T cells after activation by anti-CD 3/anti-CD 28 antibodies conjugated to Dynabeads (Invitrogen). Activated T cells and vδ1γδ T cells were incubated with a series of concentrations of anti-vδ1× PD-1 bispecific antibodies or controls (IgG control or anti-PD-1) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 58C, D.

To determine the effect of anti-vδ1×pd-1 bispecific antibodies on vδ1γδt cell activation, vδ1 TCR down-regulation was quantified in the presence of PD-1 ⁺ T cells. Briefly, anti-vδ1, anti-PD-1 and anti-vδ1×pd-1 bispecific antibodies were serially diluted in PBS and then added to assay plates. PD-1 ⁺ T cells were stained with CELLTRACE CFSE live cell stain and mixed 1:1 with skin-derived V.delta.1γ.delta.T cells or diluted 1:1 with medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4 PD-1 ⁺ T cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃ for 4 hours at 5% co ₂, then washed and stained for dead cells (eFlour, invitrogen) and vδ1 TCR (MiltenyiBiotec). Cells were washed and resuspended in Cell Fix (BD sciences). The VD1 TCR expression level was determined by Median Fluorescence Intensity (MFI), measured by flow cytometry using MACS Quant Analyzer. The results are shown in fig. 58E, F.

To assess the effect of anti-vδ1×pd-1 bispecific antibodies on activation of PD-1 ⁺ T cells, PD-1 ⁺ NFAT Jurkat cells (Promega, JA 2191) were incubated with anti-vδ1×pd-1 bispecific antibodies or controls (PD-1 monoclonal antibodies or anti RSVIgG ×anti PD-1) for 5 hours at 37 ℃, 5% co ₂. The assay was performed in the presence or absence of 1 μg/well of pre-coated recombinant vδ1 protein in opaque white 96-well plates. After 5 hours, bio-Glo luciferase reagent (Promega) was added to the cells at a 1:1 ratio. After incubation for 5 minutes at room temperature, the luminescence signal was detected on a BioTek Synergy plate reader. The original luminescence signal is converted into a Relative Luminescence Unit (RLU) multiple. The results are shown in fig. 5858G.

Conclusion: anti-vδ1×pd-1 bispecific antibodies enhance activation of vδ1γδ T cells, for example, by cross-linking via PD-1 ⁺ CD4 or CD 8T cells, and blocking PD-1/PD-L1 immune checkpoint inhibition in CD4 or CD 8T cells.

Example 36: vδ1-4-1BB bispecific antibodies enhance vδ1γδ T cell and CD 8T cell activation

Binding kinetics of anti-V.delta.1 (ADT 1-4-2), anti-4-1 BB (based on Wu Tuolu mab) and anti-V.delta.1.times.4-1 BB (ADT 1-4-2.times. Wu Tuolu mab) (SEQ ID NO:407 or SEQ ID NO:414 and SEQ ID NO: 418) antibodies to their targets (i.e., V.delta.1 chains and 4-1BB of γδ TCR) were established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human V.delta.1 heterodimer or human 4-1BB flowed through cells at the highest concentration of 100 nM. The results are shown in fig. 59A. The KD values are as follows:

to assess dual binding of the bispecific antibody to both target ligands, recombinant 4-1BB was first immobilized on a carboxymethyl dextran sensor chip (Reichert Technologies) at 10ug/ml and then flowed through the bispecific antibody at 100 nM. The ability to subsequently bind vδ1γδtcr was then assessed by flowing 100nM through recombinant human vδ1 heterodimer. All experiments were performed at room temperature. The results are shown in fig. 59B.

The binding of anti-vδ1×4-1BB bispecific antibodies to vδ1γδ T cells and 4-1BB ⁺ immune cells was assessed by flow cytometry. Initially, CD8 ⁺ T cells were negatively selected by magnetic sorting through PBMC buffy coats extracted from whole blood. Upon activation by the anti-CD 3/anti-CD 28 antibody conjugated to Dynabeads (Invitrogen), the cell surface expression of 4-1BB on CD 8T cells was elevated. Activated 4-1BB ⁺ CD 8T cells and V.delta.1γ.delta.T cells were incubated with a series of concentrations of anti-V.delta.1X1.4-1 BB bispecific antibodies or controls (IgG control or anti-4-1 BB) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 59C, D.

To determine the effect of anti-vδ1x4-1 BB bispecific antibodies on vδ1γδt cell activation, vδ TCR down-regulation was quantified in the presence of 4-1BB ⁺ T cells. Briefly, anti-V.delta.1, anti-4-1 BB and anti-V.delta.1X 4-1BB bispecific antibodies were serially diluted in PBS and then added to the assay plate. 4-1BB ⁺ CD 8T cells were stained with CELLTRACE CFSE vital cell stain and mixed 1:1 with skin-derived V.delta.1γ.delta.T cells or diluted 1:1 with medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4-1 BB ⁺ CD 8T cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃ for 4 hours at 5% co ₂, then washed and stained for dead cells (eFlour, invitrogen) and vδ1TCR (MiltenyiBiotec). Cells were washed and resuspended in Cell Fix (BD sciences). The VD 1TCR expression level was determined by Median Fluorescence Intensity (MFI), measured by flow cytometry using MACS Quant Analyzer. The results are shown in fig. 59E, F.

To evaluate the effect of anti-vδ1×4-1BB bispecific antibody on activation of 4-1BB ⁺ T cells, 4-1BB ⁺ NFAT Jurkat cells (Promega, JA 2191) were incubated with anti-vδ1×4-1BB bispecific antibody or control (anti-4-1 BB monoclonal antibody or anti-RSVIgG ×anti-4-1 BB) for 5 hours at 37 ℃, 5% co ₂. The assay was performed in the presence or absence of 1 μg/well of pre-coated recombinant vδ1 protein in opaque white 96-well plates. After 5 hours, bio-Glo luciferase reagent (Promega) was added to the cells at a 1:1 ratio. After incubation for 5 minutes at room temperature, the luminescence signal was detected on a BioTek Synergy plate reader. The original luminescence signal is converted into a Relative Luminescence Unit (RLU) multiple. The result is shown in fig. 59G.

Conclusion: anti-vδ1x4-1 BB bispecific antibodies enhance activation of vδ1γδ T cells, for example, by cross-linking with 4-1BB ⁺ CD 8T cells, and activating 4-1BB ⁺ T cells.

Example 37: vδ1-OX40 bispecific antibodies enhance vδ1γδ T cell and CD 4T cell activation

Binding kinetics for binding of anti-V.delta.1 (ADT 1-4-2), anti-OX 40 (based on Percoll) and anti-V.delta.1X OX40 (ADT 1-4-2X Percoll) (SEQ ID NO:409 or SEQ ID NO:414 and SEQ ID NO: 419) antibodies to their targets (i.e., V.delta.1 chain of γδ TCR and OX 40) were established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human vδ1 heterodimer or human OX40 flowed through cells at the highest concentration of 100 nM. The results are shown in fig. 60A. The KD values are as follows:

To assess dual binding of the bispecific antibody to both target ligands, recombinant OX40 was first immobilized on a carboxymethyl dextran sensor chip (Reichert Technologies) at 10ug/ml and then flowed through the bispecific antibody at 100 nM. The ability to subsequently bind vδ1γδtcr was then assessed by flowing 100nM through recombinant human vδ1 heterodimer. All experiments were performed at room temperature. The results are shown in fig. 60B.

Binding of anti-vδ1xox 40 bispecific antibodies to vδ1γδ T cells and OX40 ⁺ immune cells was assessed by flow cytometry. Initially, CD4 ⁺ T cells were negatively selected by magnetic sorting through PBMC buffy coats extracted from whole blood. After activation by the anti-CD 3/anti-CD 28 antibody conjugated to Dynabeads (Invitrogen), OX40 expression on the cell surface on CD 4T cells was elevated. Activated OX40 ⁺ CD 4T cells and vδ1γδ T cells were incubated with a series of concentrations of anti-vδ1xox 40 bispecific antibodies or controls (IgG control or anti-OX 40) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 60C, D.

To determine the effect of anti-vδ1×ox40 bispecific antibodies on vδ1γδt cell activation, vδ1 TCR down-regulation was quantified in the presence of OX40 ⁺ T cells. Briefly, anti-vδ1, anti-OX 40 and anti-vδ1×ox40 bispecific antibodies were serially diluted in PBS and then added to assay plates. OX40 ⁺ CD 4T cells were stained with CELLTRACE CFSE viable cell stain and mixed 1:1 with skin-derived vδ1γδ T cells or diluted 1:1 with culture medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4 OX40 ⁺ CD 4T cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃ for 4 hours at 5% co ₂, then washed and stained for dead cells (eFlour, invitrogen) and vδ1 TCR (MiltenyiBiotec). Cells were washed and resuspended in Cell Fix (BD sciences). The VD1 TCR expression level was determined by Median Fluorescence Intensity (MFI), measured by flow cytometry using MACS Quant Analyzer. The results are shown in fig. 60E, F.

To evaluate the effect of anti-vδ1×ox40 bispecific antibodies on activation of OX40 ⁺ T cells, OX40 ⁺ NFAT Jurkat cells (Promega, JA 2191) were incubated with anti-vδ1×ox40 bispecific antibodies or controls (OX 40L (OX 40 ligand), anti-OX 40 or anti-RSVIgG ×anti-OX 40) for 5 hours at 37 ℃ at 5% co ₂. The assay was performed in the presence or absence of 1 μg/well of pre-coated recombinant vδ1 protein in opaque white 96-well plates. After 5 hours, bio-Glo luciferase reagent (Promega) was added to the cells at a 1:1 ratio. After incubation for 5 minutes at room temperature, the luminescence signal was detected on a BioTek Synergy plate reader. The original luminescence signal is converted into a Relative Luminescence Unit (RLU) multiple. The results are shown in fig. 60G.

Conclusion: anti-vδ1xox 40 bispecific antibodies enhance the activation of vδ1γδ T cells, for example, by cross-linking with OX40 ⁺ CD 4T cells, and activating OX40 ⁺ T cells.

Example 38: vδ1-TIGIT bispecific antibodies enhance vδ1 γδ T cell activation and block TIGIT/PVR (CD 155) checkpoint inhibition

Binding kinetics for binding of anti-V.delta.1 (ADT 1-4-2), anti-TIGIT (based on the Tiaret Li Youshan antibody) and anti-V.delta.1X TIGIT (ADT 1-4-2X Tiaret Li Youshan antibody) (SEQ ID NO:411 or SEQ ID NO:439 and SEQ ID NO: 420) antibodies to their targets (i.e., V.delta.1 chain of γδ TCR and TIGIT) were established by SPR analysis using a Reichert 4SPR instrument (Reichert Technologies). Antibody (1.5 ug/mL) was coated onto Planar protein A sensor chip (Reichert Technologies), increasing baseline by approximately 500: 500uRIU. Recombinant human vδ1 heterodimer or human TIGIT flowed through the cells at the highest concentration of 100 nM. The results are shown in fig. 61A. The KD values are as follows:

To assess dual binding of the bispecific antibody to both target ligands, recombinant TIGIT was first immobilized on a carboxymethyl dextran sensor chip (Reichert Technologies) at 10ug/ml and then flowed through the bispecific antibody at 100 nM. The ability to subsequently bind vδ1γδtcr was then assessed by flowing 100nM through recombinant human vδ1 heterodimer. All experiments were performed at room temperature. The results are shown in fig. 61B.

Binding of anti-vδ1×tigit bispecific antibodies to vδ1γδ T cells and TIGIT ⁺ immune cells was assessed by flow cytometry. Initially, CD4 and CD 8T cells were negatively selected by magnetic sorting through PBMC buffy coats extracted from whole blood. Cell surface expression of 4-1BB was detected on CD4 and CD 8T cells after activation by anti-CD 3/anti-CD 28 antibodies conjugated to Dynabeads (Invitrogen). Activated T cells and V.delta.1γ.delta.T cells were incubated with a series of concentrations of anti V.delta.1X4-1 BB bispecific antibodies or controls (IgG control or anti-4-1 BB) for 15 minutes. After washing, cells were incubated with anti-human IgG secondary for an additional 15 minutes, then washed and fixed. The amount of antibody bound to each cell type was determined by flow cytometry. The results are shown in fig. 61C, D.

To determine the effect of anti-vδ1×tigit bispecific antibodies on vδ1γδt cell activation, vδ1 TCR down-regulation was quantified in the presence of TIGIT ⁺ T cells. Briefly, anti-vδ1, anti-TIGIT, and anti-vδ1×tigit bispecific antibodies were serially diluted in PBS and then added to assay plates. TIGIT ⁺ CD 8T cells were stained with CELLTRACE CFSE viable cell stain and mixed 1:1 with skin-derived vδ1γδ T cells or diluted 1:1 with culture medium. The cell suspension was seeded into assay plates with 2.5X10-4 V.delta.1γ.delta.T cells per well with or without 2.5X10-4 TIGIT ⁺ CD 8T cells per well. The final antibody concentration was determined to be in the range of 200nM to 2 pM. Cells were incubated at 37 ℃ for 4 hours at 5% co ₂, then washed and stained for dead cells (eFlour, invitrogen) and vδ1 TCR (MiltenyiBiotec). Cells were washed and resuspended in Cell Fix (BD sciences). The VD1 TCR expression level was determined by Median Fluorescence Intensity (MFI), measured by flow cytometry using MACS Quant Analyzer. The results are shown in fig. 61E, F.

To evaluate the effect of anti-vδ1×tigit bispecific antibodies on activation of TIGIT ⁺ T cells, TIGIT ⁺ NFAT Jurkat cells (Promega, JA 2191) were incubated with anti-vδ1×tigit bispecific antibodies or controls (anti-TIGIT monoclonal antibodies or anti-RSVIgG ×anti-TIGIT) for 5 hours at 37 ℃, 5% co ₂. The assay was performed in the presence or absence of 1 μg/well of pre-coated recombinant vδ1 protein in opaque white 96-well plates. After 5 hours, bio-Glo luciferase reagent (Promega) was added to the cells at a 1:1 ratio. After incubation for 5 minutes at room temperature, the luminescence signal was detected on a BioTek Synergy plate reader. The original luminescence signal is converted into a Relative Luminescence Unit (RLU) multiple. The results are shown in fig. 61G.

Conclusion: anti-vδ1×tigit bispecific antibodies enhance activation of vδ1γδ T cells by cross-linking via TIGIT ⁺ CD 8T cells, and blocking TIGIT/CD155 immune checkpoint inhibition in CD 8T cells.

Example 39: anti-vδ1 antibodies do not induce ADCC

ADCC reporter bioassay (Promega) was used to assess the level of ADCC (antibody dependent cell-mediated cytotoxicity) induced by the anti-vδ1 antibodies compared to the control antibodies.

ADCC refers to a biological phenomenon in which effector cells kill target cells labeled with antibodies. Effector cells bind to the antibody through their fcγ receptor, and subsequently kill the target cells. The ADCC reporter bioassay presented herein reveals the potential ADCC mechanism of action of antibodies tested in the assay by detecting early initiation of ADCC via NFAT (nuclear factor activating T cells) pathway activation gene transcription. The reporter assay is an engineered system that is further engineered to induce further firefly luciferase activation upon activation thereof using effector cells (Jurkat) expressing a high affinity fcyriiia receptor linked to the NFAT pathway. Luciferase activity was quantified with a luminescence reading that can be correlated with the level of ADCC that occurred.

This assay is used to see if an anti-vδ1 antibody or, suitably, an anti-vδ1 arm of a multispecific antibody would drive the ADCC response. The target cells used were γδ cells bound to anti-vδ1 antibodies by vδ1γδ TCR. If an ADCC mechanism is present, the anti-vδ1 antibody will bind to the Fc gamma receptor on the assay effector cell and generate a luminescent signal; if no signal is generated, it indicates that ADCC has not occurred.

ADCC reporter bioassay kit (Promega) was used for this assay. One bottle of Bio-Glo luciferase assay buffer was thawed and transferred to a substrate bottle. The mixture was kept at room temperature for 4-6 hours. Dilution plates were prepared for the following antibodies, with antibody concentrations (3X concentration) ranging from 10nM to 0.01nM (final concentration): anti-vδ1 antibody (ADT 1-4-2), the same anti-vδ1 antibody but Fc-depleted (L235A, G237A) (ADT 1-4-2 LAGA), rituximab, RSV, and OKT3. Target cells (γδ cells) were seeded at 25 μl per well into 2 assay plates. Then 25 μl of the appropriate antibody solution from the antibody dilution plate was transferred into the appropriate wells. Effector cells (engineered Jurkat cells) were thawed into warm assay buffer, resuspended into 4ml assay buffer, and 25 μl effector cell solution was removed into each well. The plates were then incubated at 37℃for 4.5 hours. After the incubation period, the plates were allowed to equilibrate to room temperature, then 75 μl of Bio-Glo luciferase assay reagent was added to each well, and the plates were incubated at room temperature for 10 minutes. The plate was then read using a Biotek H4 plate reader, which collects the luminescence signal from the plate (as relative light unit RLU). The fold induction was calculated using the following equation: fold induction = RLU (induction-background)/RLU (no antibody control-background).

OKT3 (anti-CD 3 antibody) was used as a positive control. As an additional positive control Raji cells, but not γδ cells, were seeded into control wells. Raji cells are a widely accepted cell line for exhibiting a strong ADCC response when used with the anti-CD 20 antibody rituximab. As an internal control and to see that in the absence of vδ1 binding on γδ cells, the anti-vδ1 antibodies of the present invention and the same but Fc-failed anti-vδ1 antibody (L235A, G237A) were also added with effector cells alone.

The results are shown in fig. 62.

Conclusion: the use of rituximab with Raji cells showed a strong ADCC reaction in the positive control and even stronger ADCC reaction under OKT3 against γδ cells. In contrast, no ADCC response was detected under either condition using the anti-vδ1 antibodies of the present invention, the same but Fc-disabled anti-vδ1 antibody L235A, G a or the RSV negative control. This suggests that in this system, antibodies of the invention that bind to vδ1 (e.g. anti-vδ1mAb or anti-vδ1 multispecific antibodies) do not show evidence of ADCC mechanisms of action. Notably, even Fc-effective anti-vδ1 antibodies do not deplete γδ T cells, which provides an option to maintain Fc function in the anti-vδ1 antibodies presented herein, increasing functionality, e.g. in a high fcγ tumor environment. This also highlights that such anti-vδ1 antibodies are suitable for inclusion in bispecific antibody formats as described herein.

Suitably, the multispecific antibodies of the invention that additionally specifically bind to the second antigen share the functional properties of antibodies provided in a monospecific format.

Description of the embodiments

The present invention includes at least the following numbered embodiments:

1. an anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 55 to 78 and 133 to 143, having an amino acid sequence of or consisting of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 82 to 105 and 147 to 157 has an amino acid sequence of or a VLCDR3 consisting of at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity.

2. An anti-TCR delta variable 1 (anti-vδ1) antibody of embodiment 1, or an antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, the VHCDR1 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO: 51. 52 and 130;

A VHCDR2, the VHCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:53 and 131; and

A VHCDR3 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:55 to 78 and 133 to 143; and/or

A light chain variable region comprising:

A VLCDR1, the VLCDR1 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:79 and 144;

A VLCDR2 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:80 and 145; and

A VLCDR3 comprising or consisting of an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:82 to 105 and 147 to 157.

3. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of embodiment 1 or embodiment 2, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 2 to 25 and 107 to 117 has or consists of a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 27 to 50 and 119 to 129 has or consists of a sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

4. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a VHCDR3, said VHCDR3 comprising a nucleotide sequence identical to SEQ ID NO:55 to 78, or consists of an amino acid sequence having at least 90% sequence identity; and/or

VLCDR3, said VLCDR3 comprising an amino acid sequence identical to SEQ ID NO:82 to 105, or consists of an amino acid sequence having at least 90% sequence identity.

5. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising:

a VHCDR3, said VHCDR3 comprising the amino acid sequence of SEQ ID NO:55 to 78 or consists of the amino acid sequence of any one of claims; and/or

VLCDR3, said VLCDR3 comprising SEQ ID NO:82 to 105 or consists of the amino acid sequence of any one of the amino acid sequences

6. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, further comprising:

a VHCDR1, said VHCDR1 comprising a nucleotide sequence identical to SEQ ID NO:51 or SEQ ID NO:52 or consists of an amino acid sequence having at least 90% sequence identity;

VHCDR2, said VHCDR2 comprising a nucleotide sequence identical to SEQ ID NO:53 or consists of an amino acid sequence having at least 90% sequence identity;

VLCDR1, said VLCDR1 comprising a nucleotide sequence identical to SEQ ID NO:79 or consists of an amino acid sequence having at least 90% sequence identity; and/or

A VLCDR2, said VLCDR2 comprising a nucleotide sequence identical to SEQ ID NO:80 or consists of an amino acid sequence having at least 90% sequence identity.

7. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, further comprising:

a VHCDR1, said VHCDR1 comprising the amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52 or consists of said amino acid sequence;

VHCDR2, said VHCDR2 comprising the amino acid sequence of SEQ ID NO:53 or consists of said amino acid sequence;

VLCDR1, said VLCDR1 comprising SEQ ID NO:79 or consists of the amino acid sequence of seq id no; and/or

A VLCDR2, said VLCDR2 comprising SEQ ID NO:80 or consists of said amino acid sequence.

8. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising:

a. comprising SEQ ID NOs: 51. 53 and 55 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 82 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

b. comprising SEQ ID NOs: 51. 53 and 56 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 83 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

c. comprising SEQ ID NOs: 51. 53 and 57 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 84, or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

d. comprising SEQ ID NOs: 51. 53 and 58 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 85 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

e. Respectively comprises SEO ID NO: 51. 53 and 59 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 86 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

f. Comprising SEQ ID NOs: 51. 53 and 60 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 87 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

g. Comprising SEQ ID NOs: 52. 53 and 61 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 88 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

h. comprising SEQ ID NOs: 51. 53 and 62 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 89 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

i. Comprising SEQ ID NOs: 51. 53 and 63 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 90 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

j. comprising SEQ ID NOs: 51. 53 and 64 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 91 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

k. Comprising SEQ ID NOs: 51. 53 and 65 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 92 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

l. comprising SEQ ID NO: 52. 53 and 66 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 93, or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

m. comprising SEQ ID NO: 51. 53 and 67 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 94 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

n. comprising SEQ ID NO: 51. 53 and 68 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 95 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

o. comprising SEQ ID NO: 51. 53 and 69 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 96 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

p. comprising SEQ ID NO: 51. 53 and 70 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 79. 80 and 97 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

q. comprises the amino acid sequence of SEQ ID NO: 51. 53 and 71 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 98 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

r. comprising SEQ ID NO: 51. 53 and 72 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 99 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

s. comprising SEQ ID NO: 51. 53 and 73 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 100 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

t. comprises the amino acid sequence of SEQ ID NO: 52. 53 and 74 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 101 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

u. comprising SEQ ID NO: 51. 53 and 75 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 102 or VLCDR1, VLCDR2, and VLCDR3 consisting of said amino acid sequences;

v. comprising SEQ ID NOs: 51. 53 and 76 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 103 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

w. comprises SEQ ID NO: 51. 53 and 77 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 104 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences; or (b)

X. comprises SEQ ID NOs: 51. 53 and 78 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 105 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences.

9. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising

A VH comprising a sequence identical to SEQ ID NO:2 to 25, an amino acid sequence having at least 90% sequence identity; and/or

VL comprising a sequence identical to SEQ ID NO:27 to 50, or consists of an amino acid sequence having at least 90% sequence identity.

10. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising:

a VH comprising SEQ ID NO:2 to 25 or consists of the amino acid sequence of any one of claims 2 to 25; and/or

VL comprising the amino acid sequence of SEQ ID NO:27 to 50 or consists of the amino acid sequence of any one of claims.

11. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising:

a. Comprising a sequence identical to SEQ ID NO:2 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:27 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

b. comprising a sequence identical to SEQ ID NO:3 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:28 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

c. Comprising a sequence identical to SEQ ID NO:4 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:29 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

d. Comprising a sequence identical to SEQ ID NO:5 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:30 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

e. comprising a sequence identical to SEQ ID NO:6 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:31 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

f. Comprising a sequence identical to SEQ ID NO:7 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:32 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

g. Comprising a sequence identical to SEQ ID NO:8 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:33 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

h. Comprising a sequence identical to SEQ ID NO:9 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:34 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

i. Comprising a sequence identical to SEQ ID NO:10 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:35 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

j. comprising a sequence identical to SEQ ID NO:11 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:36 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

k. Comprising a sequence identical to SEQ ID NO:12 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:37 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

l. comprising a sequence identical to SEQ ID NO:13 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:38 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

m. comprises a sequence identical to SEQ ID NO:14 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:39 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

n. comprises a sequence identical to SEQ ID NO:15 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:40 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

o. comprising a sequence identical to SEQ ID NO:16 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:41 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

p. comprising a sequence identical to SEQ ID NO:17 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:42 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

q. comprises a sequence identical to SEQ ID NO:18 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:43 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

r. comprises a sequence identical to SEQ ID NO:19 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:44 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

s. comprises a sequence identical to SEQ ID NO:20 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:45 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

t. comprises a sequence identical to SEQ ID NO:21 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:46 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

u. comprises a sequence identical to SEQ ID NO:22 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:47 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

v. comprising a sequence identical to SEQ ID NO:23 and a VH comprising or consisting of an amino acid sequence having at least 90% sequence identity to SEQ ID NO:48 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

w. comprises a sequence identical to SEQ ID NO:24 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:49 or a VL consisting of an amino acid sequence having at least 90% sequence identity; or (b)

X. comprises a sequence identical to SEQ ID NO:25 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:50 or a VL consisting of an amino acid sequence having at least 80% sequence identity.

12. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, comprising:

a. comprising SEQ ID NO:2 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:27 or a VL consisting of said amino acid sequence;

b. comprising SEQ ID NO:3 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:28 or a VL consisting of said amino acid sequence;

c. Comprising SEQ ID NO:4 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:29 or a VL consisting of said amino acid sequence;

d. comprising SEQ ID NO:5 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:30 or a VL consisting of said amino acid sequence;

e. Comprising SEQ ID NO:6 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:31 or a VL consisting of said amino acid sequence;

f. Comprising SEQ ID NO:7 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:32 or a VL consisting of said amino acid sequence;

g. Comprising SEQ ID NO:8 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:33 or a VL consisting of said amino acid sequence;

h. Comprising SEQ ID NO:9 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:34 or a VL consisting of said amino acid sequence;

i. comprising SEQ ID NO:10 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:35 or a VL consisting of said amino acid sequence;

j. comprising SEQ ID NO:11 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:36 or a VL consisting of said amino acid sequence;

k. Comprising SEQ ID NO:12 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:37 or a VL consisting of said amino acid sequence;

l. comprising SEQ ID NO:13 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:38 or a VL consisting of said amino acid sequence;

m. comprising SEQ ID NO:14 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:39 or a VL consisting of said amino acid sequence;

n. a polypeptide comprising SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence;

o. comprising SEQ ID NO:16 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:41 or a VL consisting of said amino acid sequence;

p. comprising SEQ ID NO:17 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:42 or a VL consisting of said amino acid sequence;

q. a polypeptide comprising SEQ ID NO:18 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:43 or a VL consisting of said amino acid sequence;

r. comprising SEQ ID NO:19 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:44 or a VL consisting of said amino acid sequence;

s. comprising SEQ ID NO:20 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:45 or a VL consisting of said amino acid sequence;

t. comprises SEQ ID NO:21 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:46 or a VL consisting of said amino acid sequence;

u. comprising SEQ ID NO:22 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:47 or a VL consisting of said amino acid sequence;

v. a polypeptide comprising SEQ ID NO:23 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:48 or a VL consisting of said amino acid sequence;

w. comprises SEQ ID NO:24 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:49 or a VL consisting of said amino acid sequence; or (b)

X. comprises SEQ ID NO:25 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:50 or a VL consisting of said amino acid sequence.

13. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof comprises an amino acid residue other than serine at position 74 according to the IMGT kappa light chain variable sequence numbering system.

14. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 13, wherein said residue at position 74 is an amino acid according to the IMGT numbering system selected from the group consisting of: glycine, alanine, valine, methionine, leucine and isoleucine.

15. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 13, wherein said residue at position 74 is an amino acid according to the IMGT numbering system selected from the group consisting of: arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine and valine.

16. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 13, wherein said residue at position 74 is an amino acid selected from the group consisting of: glycine, valine, methionine, leucine and isoleucine.

17. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 13, wherein said residue at position 74 is a leucine residue according to the IMGT numbering system.

18. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof is an antibody ：ADT1-4-105、ADT1-4-107、ADT1-4-110、ADT1-4-112、ADT1-4-117、ADT1-4-19、ADT1-4-21、ADT1-4-31、ADT1-4-139、ADT1-4-4、ADT1-4-143、ADT1-4-53、ADT1-4-173、ADT1-4-2、ADT1-4-8、ADT1-4-82、ADT1-4-83、ADT1-4-3、ADT1-4-84、ADT1-4-86、ADT1-4-95、ADT1-4-1、ADT1-4-6 and ADT1-4-138 selected from the group consisting of.

19. The anti-vδ1 antibody of embodiment 18 or an antigen binding fragment thereof, wherein said antibody comprises 1 to 10, 1 to 5 or 1 to 2 amino acid substitutions in all 6 CDR regions.

20. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 18 or embodiment 19, wherein said antibody comprises up to 2 or up to 1 amino acid substitutions in all 6 CDR regions.

21. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 18 to 20, wherein said antibody comprises 1 or 2 amino acid substitutions in all 6 CDR regions.

22. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 18 to 21, wherein the antibody comprises 1 to 10, 1 to 5 or 1 to 2 amino acid substitutions in one or more framework regions.

23. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 18 to 22, wherein said antibody comprises up to 2 or up to 1 amino acid substitutions in one or more framework regions.

24. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 18 to 23, wherein said antibody comprises 1 or 2 amino acid substitutions in one or more framework regions.

25. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 18 to 24, wherein said amino acid substitution is a conservative amino acid substitution.

26. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

an HCDR1 sequence comprising or consisting of a sequence of GDSVSSKSX ₁ a;

An HCDR2 sequence comprising SEQ ID NO:53 or consists of said sequence

An HCDR3 sequence comprising or consisting of a sequence of X ₂WX₃X₄X₅X₆DX₇, wherein the HCDR3 sequence is not SEQ ID NO:54;

an LCDR1 sequence comprising SEQ ID NO:79 or consists of said sequence;

An LCDR2 sequence comprising SEQ ID NO:80 or consists of said sequence; and

An LCDR3 sequence comprising or consisting of a sequence of QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T, wherein the LCDR3 sequence is not SEQ ID NO:81;

wherein each of X1 to X13 is a naturally occurring amino acid.

27. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 26, wherein:

a.X ₁ is selected from the group consisting of: a and V;

b.X ₂ is selected from the group consisting of: s and T;

c.X ₃ is selected from the group consisting of: v, A and L;

d.X ₄ is selected from the group consisting of: G. e and D;

e.X ₅ is selected from the group consisting of: y and N;

f.X ₆ is selected from the group consisting of: v, A and P;

g.X ₇ is selected from the group consisting of: v, Y and R;

h.X ₈ is selected from the group consisting of: K. r and G;

i.X ₉ is selected from the group consisting of: s and K;

j.X ₁₀ is selected from the group consisting of: t, Q, A, E and D;

k.X ₁₁ is selected from the group consisting of: p, H and D;

l.X ₁₂ is selected from the group consisting of: q, R, K, W, P, E and I; and

M.X ₁₃ is selected from the group consisting of: I. v and L.

28. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 26 or embodiment 27, wherein said antibody further comprises:

an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence;

An HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence;

an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence;

HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence;

An LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence;

an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence;

LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or 178 or consists of said sequence; and

An LFR4 sequence, said LFR4 sequence comprising the sequence of SEQ ID NO: 179. 180, 181 or 182 or a sequence consisting of said sequences.

29. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof:

a. Binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

b. Optionally binds to cynomolgus monkey TRDV1 (SEQ ID NO 308) with a binding affinity (K _D, e.g. measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

30. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof:

a. binds to human TRDV1 with a KD of less than about 100nM (preferably less than about 50 nM);

b. Binds to cynomolgus TRDV1 with a KD of less than about 100nM (preferably less than about 50 nM);

c. The IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM) and the IC90 for TCR downregulation is less than about 100nM (less than about 50 nM);

d. IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM)

E. the IC90 for killing THP-1 cells is less than about 50nM.

31. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 26, wherein:

a.X ₁ is selected from the group consisting of: a and V;

b.X ₂ is selected from the group consisting of: s and T;

c.X ₃ is selected from the group consisting of: v, A and L;

d.X ₄ is selected from the group consisting of: G. e and D;

e.X ₅ is selected from the group consisting of: y and N;

f.X ₆ is selected from the group consisting of: v, A and P;

g.X ₇ is selected from the group consisting of: v, Y and R;

h.X ₈ is selected from the group consisting of: K. r and G;

i.X ₉ is selected from the group consisting of: s and K;

j.X ₁ 0 is selected from the group consisting of: t, Q, A, E and D;

k.X ₁₁ is selected from the group consisting of: p and H;

l.X ₁₂ is selected from the group consisting of: q, R, K, W, P, E and I; and

M.X ₁₃ is selected from the group consisting of: I. v and L.

32. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 31, wherein said antibody further comprises:

an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence;

An HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence;

an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence;

HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence;

An LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence;

an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence;

LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or consists of said sequence; and

An LFR4 sequence, said LFR4 sequence comprising the sequence of SEQ ID NO: 179. 180, 181 or 182 or a sequence consisting of said sequences.

33. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM, preferably less than about 5nM,

B. binds to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

34. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 26, wherein:

a.X ₁ is selected from the group consisting of: a and V;

b.X ₂ is selected from the group consisting of: s and T;

c.X ₃ is selected from the group consisting of: v, A and L;

d.X ₄ is selected from the group consisting of: g and D;

e.X ₅ is selected from the group consisting of: y and N;

f.X ₆ is selected from the group consisting of: v, A and P;

g.X ₇ is selected from the group consisting of: v, Y and R;

h.X ₈ is selected from the group consisting of: k and R;

i.X ₉ is selected from the group consisting of: s and K;

j.X ₁ 0 is selected from the group consisting of: t, Q, A and E;

k.X ₁₁ is selected from the group consisting of: p and H;

l.X ₁₂ is selected from the group consisting of: q, K, W, P and I; and

M.X ₁₃ is selected from the group consisting of: v and L.

35. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 34, wherein said antibody further comprises:

an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:170 or 171 or consists of said sequence;

An HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:172 or consists of said sequence;

an HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:173 or consists of said sequence;

HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:174 or consists of said sequence;

An LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:175 or consists of said sequence;

an LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:176 or consists of said sequence;

LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:177 or consists of said sequence; and

An LFR4 sequence, said LFR4 sequence comprising the sequence of SEQ ID NO:179 or 181 or consists of said sequence.

36. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 1 nM),

B. Binding to cynomolgus monkey TRDV1 with a K _D of less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

37. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 from amino acid residues 37-53 and/or 59-77.

38. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

39. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the epitope bound by said antibody or antigen binding fragment thereof consists of the amino acid sequence of SEQ ID NO:272, amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

40. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a VHCDR3, said VHCDR3 comprising a nucleotide sequence identical to SEQ ID NO:133 to 143 or consists of an amino acid sequence having at least 90% sequence identity; and/or

VLCDR3, said VLCDR3 comprising an amino acid sequence identical to SEQ ID NO:147 to 157, or consists of an amino acid sequence having at least 90% sequence identity.

41. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 40, comprising:

a VHCDR3, said VHCDR3 comprising the amino acid sequence of SEQ ID NO:133 to 143 or consists of the amino acid sequence of any one of seq id no; and/or

VLCDR3, said VLCDR3 comprising SEQ ID NO:147 to 157 or consists of an amino acid sequence of any one of claims 147 to 157.

42. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 41, further comprising:

a VHCDR1, said VHCDR1 comprising a nucleotide sequence identical to SEQ ID NO:130 or consists of an amino acid sequence having at least 90% sequence identity;

VHCDR2, said VHCDR2 comprising a nucleotide sequence identical to SEQ ID NO:131 or consists of an amino acid sequence having at least 90% sequence identity;

VLCDR1, said VLCDR1 comprising a nucleotide sequence identical to SEQ ID NO:144 or consists of an amino acid sequence having at least 90% sequence identity; and/or

A VLCDR2, said VLCDR2 comprising a nucleotide sequence identical to SEQ ID NO:145 or consists of an amino acid sequence having at least 90% sequence identity.

43. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 42, further comprising:

a VHCDR1, said VHCDR1 comprising the amino acid sequence of SEQ ID NO:130 or consists of the amino acid sequence of seq id no;

VHCDR2, said VHCDR2 comprising the amino acid sequence of SEQ ID NO:131 or consists of said amino acid sequence;

VLCDR1, said VLCDR1 comprising SEQ ID NO:144 or consists of the amino acid sequence of 144; and/or

A VLCDR2, said VLCDR2 comprising SEQ ID NO:145 or consists of said amino acid sequence.

44. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 43, comprising:

a. comprising SEQ ID NOs: 130. 131 and 133 or a VHCDR1, a VHCDR2 and a VHCDR3 consisting of said amino acid sequences, and a nucleic acid sequence comprising SEQ ID NO: 144. 145 and 147 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

b. comprising SEQ ID NOs: 130. 131 and 134 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 148 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

c. comprising SEQ ID NOs: 130. 131 and 135 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 149 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

d. Comprising SEQ ID NOs: 130. 131 and 136 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 150 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

e. Comprising SEQ ID NOs: 130. 131 and 137 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 151 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

f. Comprising SEQ ID NOs: 130. 131 and 138 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 152 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

g. Comprising SEQ ID NOs: 130. 131 and 139 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 153 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

h. comprising SEQ ID NOs: 130. 131 and 140 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 154 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

i. Comprising SEQ ID NOs: 130. 131 and 141 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 155 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences;

j. Comprising SEQ ID NOs: 130. 131 and 142 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a polypeptide comprising SEQ ID NO: 144. 145 and 156 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences; or (b)

K. comprising SEQ ID NOs: 130. 131 and 143 or VHCDR1, VHCDR2 and VHCDR3 consisting of said amino acid sequences, and a sequence comprising SEQ ID NO: 144. 145 and 157 or VLCDR1, VLCDR2 and VLCDR3 consisting of said amino acid sequences.

45. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 44, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A VH comprising a sequence identical to SEQ ID NO:107 to 117 or consists of an amino acid sequence having at least 90% sequence identity; and/or

VL comprising a sequence identical to SEQ ID NO:119 to 129, or consists of an amino acid sequence having at least 90% sequence identity.

46. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 45, comprising:

a VH comprising SEQ ID NO:107 to 117 or consists of the amino acid sequence of any one of seq id no; and/or

VL comprising the amino acid sequence of SEQ ID NO:119 to 129 or consists of the amino acid sequence of any one of claims 119 to 129.

47. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 46, comprising:

a. comprising a sequence identical to SEQ ID NO:107 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:119 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

b. Comprising a sequence identical to SEQ ID NO:108 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:120 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

c. Comprising a sequence identical to SEQ ID NO:109 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:121 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

d. Comprising a sequence identical to SEQ ID NO:110 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:122 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

e. Comprising a sequence identical to SEQ ID NO:111 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:123 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

f. Comprising a sequence identical to SEQ ID NO:112 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:124 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

g. Comprising a sequence identical to SEQ ID NO:113 and a VH comprising or consisting of an amino acid sequence having at least 90% sequence identity to SEQ ID NO:125 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

h. comprising a sequence identical to SEQ ID NO:114 and a VH comprising or consisting of an amino acid sequence having at least 90% sequence identity to SEQ ID NO:126 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

i. Comprising a sequence identical to SEQ ID NO:115 or a VH consisting of said amino acid sequence and comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:127 or a VL consisting of an amino acid sequence having at least 90% sequence identity;

j. Comprising a sequence identical to SEQ ID NO:116 or a VH consisting of said amino acid sequence having at least 90% sequence identity to SEQ ID NO:128 or a VL consisting of an amino acid sequence having at least 90% sequence identity; or (b)

K. Comprising a sequence identical to SEQ ID NO:117 or a VH consisting of or comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:129 or a VL consisting of an amino acid sequence having at least 90% sequence identity.

48. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 47, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. Comprising SEQ ID NO:107 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:119 or a VL consisting of said amino acid sequence;

b. Comprising SEQ ID NO:108 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:120 or a VL consisting of said amino acid sequence;

c. comprising SEQ ID NO:109 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:121 or a VL consisting of said amino acid sequence;

d. comprising SEQ ID NO:110 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:122 or a VL consisting of said amino acid sequence;

e. comprising SEQ ID NO:111 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:123 or a VL consisting of said amino acid sequence;

f. Comprising SEQ ID NO:112 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:124 or a VL consisting of said amino acid sequence;

g. comprising SEQ ID NO:113 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:125 or a VL consisting of said amino acid sequence;

h. Comprising SEQ ID NO:114 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:126 or a VL consisting of said amino acid sequence;

i. Comprising SEQ ID NO:115 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:127 or a VL consisting of said amino acid sequence;

j. comprising SEQ ID NO:116 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:128 or a VL consisting of said amino acid sequence; or (b)

K. Comprising SEQ ID NO:117 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:129 or a VL consisting of said amino acid sequence.

49. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 48, wherein said antibody or antigen-binding fragment thereof comprises a kappa light chain variable sequence having an amino acid residue other than serine at position 74 according to the IMGT numbering system.

50. The anti-vδ1 antibody of embodiment 49 or an antigen binding fragment thereof, wherein said residue at position 74 of said light chain variable sequence is an amino acid according to the IMGT numbering system selected from the group consisting of: glycine, alanine, valine, methionine, leucine and isoleucine.

51. The anti-vδ1 antibody of embodiment 49 or an antigen binding fragment thereof, wherein said residue at position 74 of said light chain variable sequence is an amino acid according to the IMGT numbering system selected from the group consisting of: arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine and valine.

52. The anti-vδ1 antibody of embodiment 49 or an antigen binding fragment thereof, wherein said residue at position 74 of said light chain variable sequence is an amino acid according to the IMGT numbering system selected from the group consisting of: glycine, valine, methionine, leucine and isoleucine.

53. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 49, wherein said residue at position 74 of said light chain variable sequence is a leucine residue according to the IMGT numbering system.

54. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 53, wherein said anti-vδ1 antibody or antigen-binding fragment thereof is an antibody selected from the group consisting of: ADT1-7-10, ADT1-7-15, ADT1-7-17, ADT1-7-18, ADT1-7-19, ADT1-7-20, ADT1-7-22ADT1-7-23, ADT1-7-42, ADT1-7-3, and ADT1-7-61.

55. The anti-vδ1 antibody of embodiment 54, or an antigen binding fragment thereof, wherein said antibody comprises 1 to 10, 1 to 5 or 1 to 2 amino acid substitutions in all 6 CDR regions.

56. The anti-vδ1 antibody of embodiment 54 or 55, or an antigen binding fragment thereof, wherein said antibody comprises up to 2 or up to 1 amino acid substitutions in all 6 CDR regions.

57. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 54 to 56, wherein said antibody comprises 1 or 2 amino acid substitutions in all 6 CDR regions.

58. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 54 to 57, wherein said antibody comprises 1 to 10, 1 to 5 or 1 to 2 amino acid substitutions in one or more framework regions.

59. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 54 to 55, wherein said antibody comprises up to 2 or up to 1 amino acid substitutions in one or more framework regions.

60. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 54 to 59, wherein said antibody comprises 1 or 2 amino acid substitutions in one or more framework regions.

61. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 55 to 60, wherein said amino acid substitution is a conservative amino acid substitution.

62. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

an HCDR1 sequence comprising SEQ ID NO:130 or consists of said sequence

An HCDR2 sequence comprising SEQ ID NO:131 or consists of said sequence

An HCDR3 sequence comprising or consisting of a sequence of X ₁X₂YX₃X₄ AFDI, wherein the HCDR3 sequence is not SEQ ID NO:132, a part of the material;

an LCDR1 sequence comprising SEQ ID NO:144 or consists of said sequence;

An LCDR2 sequence comprising SEQ ID NO:145 or consists of said sequence; and

An LCDR3 sequence comprising or consisting of a sequence of QQX ₅X₆X₇X₈LX₉ T, wherein the LCDR3 sequence is not SEQ ID NO:146;

wherein each of X ₁ to X ₉ is a naturally occurring amino acid.

63. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 62, wherein:

a.X ₁ is selected from the group consisting of: D. i and V;

b.X ₂ is selected from the group consisting of: d and S;

c.X ₃ is selected from the group consisting of: n, E, D, Q and A;

d.X ₄ is selected from the group consisting of: d and E;

e.X ₅ is selected from the group consisting of: t and S;

f.X ₆ is selected from the group consisting of: A. g, Y and S;

g.X ₇ is selected from the group consisting of: s and D;

h.X ₈ is selected from the group consisting of: t, E and G; and

I.X ₉ is selected from the group consisting of: l and D.

64. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 62 or embodiment 63, wherein said antibody further comprises:

an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:189 or consists of said sequence;

An HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:190 or consists of said sequence;

An HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191 or consists of said sequence;

HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192 or consists of said sequence;

an LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:193 and 194 or consists of said sequences;

An LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:195 or consists of said sequence;

LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:196 or consists of said sequence; and

An LFR4 sequence, said LFR4 sequence comprising the sequence of SEQ ID NO:197 or consists of said sequence.

65. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 64, wherein said antibody or antigen-binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

c. IC50 for killing THP-1 cells is less than about 5nM (preferably less than about 1 nM)

D. The IC90 for killing THP-1 cells is less than about 50nM.

66. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 62, wherein:

a.X ₁ is selected from the group consisting of: d and V;

b.X ₂ is selected from the group consisting of: d and S;

c.X ₃ is selected from the group consisting of: D. q and A;

d.X ₄ is selected from the group consisting of: d and E;

e.X ₅ is S;

f.X ₆ is selected from the group consisting of: a and Y;

g.X ₇ is S;

h.X ₈ is selected from the group consisting of: e and G; and

I.X ₉ is selected from the group consisting of: l and D.

67. The anti-vδ1 antibody of embodiment 66, or an antigen binding fragment thereof, wherein said antibody further comprises:

an HFR1 sequence, said HFR1 sequence comprising SEQ ID NO:189 or consists of said sequence;

An HFR2 sequence, said HFR2 sequence comprising SEQ ID NO:190 or consists of said sequence;

An HFR3 sequence, said HFR3 sequence comprising SEQ ID NO:191 or consists of said sequence;

HFR4 sequence, said HFR4 sequence comprising SEQ ID NO:192 or consists of said sequence;

An LFR1 sequence, said LFR1 sequence comprising the sequence of SEQ ID NO:193 or consists of said sequence;

An LFR2 sequence, said LFR2 sequence comprising the sequence of SEQ ID NO:195 or consists of said sequence;

LFR3 sequence, said LFR3 sequence comprising SEQ ID NO:196 or consists of said sequence; and

An LFR4 sequence, said LFR4 sequence comprising the sequence of SEQ ID NO:197 or consists of said sequence.

68. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 67, wherein said antibody or antigen-binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 5nM (preferably less than about 1 nM);

c. the IC90 for TCR downregulation is less than about 10nM (preferably less than about 5 nM);

d. IC50 for killing THP-1 cells is less than about 5nM (preferably less than about 1 nM)

E. The IC90 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM).

69. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 68, wherein the epitope bound by said antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:271 from amino acid region 5-20 and/or from 62-77.

70. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 69, wherein the epitope bound by said antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77.

71. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 40 to 70, wherein the epitope bound by said antibody or antigen-binding fragment thereof consists of the amino acid sequence of SEQ ID NO:272, amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77.

72. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises a polypeptide comprising SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, a VL sequence of the amino acid sequence of seq id no.

73. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 72, wherein said antibody is an antibody as defined in any one of embodiments 4 to 40.

74. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 4 to 40, 72 or 73, wherein said antibody or antigen binding fragment thereof:

a. binds to human TRDV1 with a KD of less than about 100nM (preferably less than about 50 nM);

b. Binds to cynomolgus TRDV1 with a KD of less than about 100nM (preferably less than about 50 nM);

c. The IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM) and the IC90 for TCR downregulation is less than about 100nM (less than about 50 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

75. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 4 to 40, 72 or 73, wherein said antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM, preferably less than about 5nM,

B. binds to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

76. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 4 to 40, 72 or 73, wherein said antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 1 nM),

B. Binding to cynomolgus monkey TRDV1 with a K _D of less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and/or

E. the IC90 for killing THP-1 cells is less than about 50nM.

77. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises a polypeptide comprising SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, a VL sequence of the amino acid sequence of seq id no.

78. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 77, wherein said antibody is an antibody as defined in any one of embodiments 41 to 71.

79. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 41 to 71, 77 or 78, wherein said antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

c. an IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM); and/or

D. The IC90 for killing THP-1 cells is less than about 50nM.

80. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 41 to 71, 77 or 78, wherein said antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 5nM (preferably less than about 1 nM);

c. the IC90 for TCR downregulation is less than about 10nM (preferably less than about 5 nM);

d. An IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM); and/or

E. The IC90 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM).

81. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-vδ1 antibody or antigen-binding fragment thereof, wherein the parent anti-vδ1 antibody or antigen-binding fragment thereof comprises:

a. Comprising SEQ ID NO:273 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:282 or a VL consisting of said amino acid sequence;

b. comprising SEQ ID NO:274 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:283 or a VL consisting of said amino acid sequence;

c. Comprising SEQ ID NO:275 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:284 or a VL consisting of said amino acid sequence;

d. Comprising SEQ ID NO:276 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:285 or a VL consisting of said amino acid sequence;

e. comprising SEQ ID NO:277 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:286 or a VL consisting of said amino acid sequence;

f. comprising SEQ ID NO:278 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:287 or a VL consisting of said amino acid sequence;

g. comprising SEQ ID NO:279 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:288 or a VL consisting of said amino acid sequence;

h. comprising SEQ ID NO:280 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:289 or a VL consisting of said amino acid sequence;

i. Comprising SEQ ID NO:281 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:290 or a VL consisting of said amino acid sequence; or (b)

J. comprising SEQ ID NO:312 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:313 or a VL consisting of said amino acid sequence.

82. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 72 to 81, wherein the affinity of said affinity matured antibody for binding to a variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) is at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 500% or more preferably at least about 1000% higher than said parent antibody, e.g. measured by Kd.

83. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 72 to 82, wherein said affinity matured anti-vδ1 antibody or antigen-binding fragment thereof comprises VH and VL sequences that are at least 80%, at least 90%, at least 95% or at least 96% identical to the corresponding parent VH and VL sequences.

84. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 72 to 82, wherein said affinity matured anti-vδ1 antibody or antigen-binding fragment comprises up to 20, such as up to 15, such as up to 10 amino acid substitutions compared to said parent antibody sequence.

85. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody does not comprise a polypeptide having a sequence of SEQ ID NO:54 and a VHCDR3 having the amino acid sequence of SEQ ID NO:81, and/or wherein the antibody does not comprise a polypeptide having the amino acid sequence of SEQ ID NO:132 and a VHCDR3 having the amino acid sequence of SEQ ID NO:146 VLCDR3.

86. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody does not comprise a polypeptide having a sequence of SEQ ID NO:1 and a VH region having SEQ ID NO:26, and/or wherein the antibody does not comprise a VL region having SEQ ID NO:106 and a VH region having SEQ ID NO: 118.

87. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof comprising a kappa light chain variable sequence, wherein the residue at position 74 is not a serine residue according to the IMGT numbering system.

88. The anti-vδ1 antibody of embodiment 85 or an antigen binding fragment thereof, wherein said residue at position 74 is an amino acid according to the IMGT numbering system selected from the group consisting of: glycine, alanine, valine, methionine, leucine and isoleucine.

89. The anti-vδ1 antibody of embodiment 88 or an antigen binding fragment thereof, wherein said residue at position 74 is an amino acid according to the IMGT numbering system selected from the group consisting of: arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine and valine.

90. The anti-vδ1 antibody of embodiment 88 or an antigen binding fragment thereof, wherein said residue at position 74 is an amino acid according to the IMGT numbering system selected from the group consisting of: glycine, valine, methionine, leucine and isoleucine.

91. The anti-vδ1 antibody of embodiment 88 or an antigen binding fragment thereof according to the IMGT numbering system, said residue at position 74 being a leucine residue.

92. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody is an IgG antibody.

93. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody is an IgG1 antibody.

94. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody is an antibody as defined in any one of embodiments 4 to 40 or 72 to 76.

95. An anti-vδ1 antibody or antigen binding fragment, wherein the antibody is a multispecific antibody that specifically binds to an epitope of:

a. a first target epitope, wherein the first target epitope is an epitope of the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR); and

B. A second target epitope.

96. An anti-vδ1 antibody or antigen binding fragment, wherein the antibody is a multispecific antibody that specifically binds to an epitope of:

a. a first target epitope, wherein the first target epitope is an epitope of the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR); and

B. A second target epitope;

Wherein the multispecific antibody comprises SEQ ID NO：385;SEQ ID NO：386;SEQ ID NO：387;SEQ ID NO：314;SEQ ID NO：393;SEQ ID NO：394;SEQ ID NO：395;SEQ ID NO：397;SEQ ID NO：388;SEQ ID NO：315;SEQ ID NO：316;SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：393;SEQ ID NO：396;SEQ ID NO：398;SEQ ID NO：399;SEQ ID NO：401;SEQ ID NO：402;SEQ ID NO：403;SEQ ID NO：404;SEQ ID NO：405;SEQ ID NO：406;SEQ ID NO：407;SEQ ID NO：408;SEQ ID NO：409;SEQ ID NO：410;SEQ ID NO：411;SEQ ID NO：412;SEQ ID NO：413;SEQ ID NO：414 and SEQ ID NO:415, a base; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:418; SEQ ID NO:505 and SEQ ID NO:415, a base; SEQ ID NO:505 and SEQ ID NO:416; SEQ ID NO:505 and SEQ ID NO:419; SEQ ID NO:505 and SEQ ID NO:418; SEQ ID NO:421 and SEQ ID NO:422, a part of the material; SEQ ID NO:504 and SEQ ID NO:422, a part of the material; SEQ ID NO:423 and SEQ ID NO: 424. SEQ ID NO:425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:429; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:414 and SEQ ID NO:435; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:434, a base; SEQ ID NO:505 and SEQ ID NO:435; SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

97. The anti-vδ1 antibody or antigen binding fragment of any one of embodiments 1 to 95, wherein said antibody is a multispecific antibody that specifically binds to an epitope of:

a. a first target epitope, wherein the first target epitope is an epitope of the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR); and

B. A second target epitope;

Wherein the multispecific antibody comprises SEQ ID NO：385;SEQ ID NO：386;SEQ ID NO：387;SEQ ID NO：314;SEQ ID NO：393;SEQ ID NO：394;SEQ ID NO：395;SEQ ID NO：397;SEQ ID NO：388;SEQ ID NO：315;SEQ ID NO：316;SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：393;SEQ ID NO：396;SEQ ID NO：398;SEQ ID NO：399;SEQ ID NO：401;SEQ ID NO：402;SEQ ID NO：403;SEQ ID NO：404;SEQ ID NO：405;SEQ ID NO：406;SEQ ID NO：407;SEQ ID NO：408;SEQ ID NO：409;SEQ ID NO：410;SEQ ID NO：411;SEQ ID NO：412;SEQ ID NO：414 and SEQ ID NO:415, a base; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:418; SEQ ID NO:505 and SEQ ID NO:415, a base; SEQ ID NO:505 and SEQ ID NO:416; SEQ ID NO:505 and SEQ ID NO:419; SEQ ID NO:505 and SEQ ID NO:418; SEQ ID NO:421 and SEQ ID NO:422, a part of the material; SEQ ID NO:504 and SEQ ID NO:422, a part of the material; SEQ ID NO:423 and SEQ ID NO: 424. SEQ ID NO:425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:429; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:414 and SEQ ID NO:435; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:434, a base; SEQ ID NO:505 and SEQ ID NO:435; SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

98. The anti-vδ1 antibody or antigen binding fragment of embodiment 94 or embodiment 95, wherein said multispecific antibody comprises SEQ ID NO：388;SEQ ID NO：393;SEQ ID NO：394;SEQ ID NO：395;SEQ ID NO：396;SEQ ID NO：397;SEQ ID NO：398;SEQ ID NO：399;SEQ ID NO：401;SEQ ID NO：402;SEQ ID NO：403;SEQ ID NO：404;SEQ ID NO：405;SEQ ID NO：406;SEQ ID NO：407;SEQ ID NO：408;SEQ ID NO：409;SEQ ID NO：410;SEQ ID NO：411;SEQ ID NO：412;SEQ ID NO：414 and SEQ ID NO:415, a base; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:418; SEQ ID NO:505 and SEQ ID NO:415, a base; SEQ ID NO:505 and SEQ ID NO:416; SEQ ID NO:505 and SEQ ID NO:419; SEQ ID NO:505 and SEQ ID NO:418; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:434, a base; SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

99. The anti-vδ1 antibody or antigen binding fragment of any one of embodiments 96-98, wherein said first target epitope is an epitope as defined in any one of embodiments 37-39 or 69-71.

100. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, that specifically binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) and competes with an antibody or antigen-binding fragment thereof of any preceding embodiment for binding to the variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR).

101. The anti-vδ1 antibody or antigen binding fragment of any preceding embodiment, wherein the antibody is Fc effective.

102. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof characterized in that it:

a. Causing the TCR on vδ1T cells to which it binds to down-regulate;

b. does not exhibit CDC or ADCC; and

C. The vδ1T cells were not depleted.

103. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of embodiment 102, wherein the antibody causes less than about 30%, or less than about 20%, or less than about 10% of a population of viable vδ T + cells via ADCC and/or CDC.

104. The anti-TCR delta variable 1 (anti-vδ1) antibody of embodiment 102, or an antigen-binding fragment thereof, wherein the antibody does not induce secretion of IL-17A.

105. The anti-TCR delta variable 1 (anti-vδ1) antibody of embodiment 102, or antigen-binding fragment thereof, wherein the antibody induces less than about 30%, or less than about 20%, or less than about 10% of IL-17A secretion induced by an equivalent CD3 antibody.

106. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 102-105, wherein the antibody or antigen-binding fragment thereof binds to human TRVD1 with a KD of less than about 10 nM.

107. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 102-106, wherein the antibody or antigen-binding fragment thereof binds to human TRVD1 with a KD of less than about 5 nM.

108. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 102-107, wherein said antibody or antigen-binding fragment thereof binds to cynomolgus monkey TRVD1 with a KD of less than about 100nM.

109. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiment 108, wherein said antibody or antigen-binding fragment thereof binds to cynomolgus monkey TRVD1 with a KD of less than about 50 nM.

110. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, that specifically binds to a variable δ1 (vδ1) chain of a human γδ T Cell Receptor (TCR) with a KD of less than about 10 nM.

111. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 110, wherein said anti-vδ1 antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof as defined in any one of embodiments 1 to 109.

112. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen binding fragment thereof binds to a variable δ1 (vδ1) chain of a cynomolgus monkey γδ T Cell Receptor (TCR) with a KD of less than about 100nM or less than about 50 nM.

113. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof down-regulates a TCR with an IC50 of less than about 50nM, preferably less than about 10nM.

114. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof down-regulates a TCR with an IC90 of less than about 100nM, preferably less than about 50nM.

115. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof kills THP-1 cells with an IC50 of less than about 10nM, preferably less than about 5nM.

116. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof kills THP-1 cells with an IC90 of less than about 50nM.

117. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody or antigen binding fragment thereof stimulates vδ1T cell proliferation.

118. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof binds human TVRD1 with a KD of less than about 100 nM.

119. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof binds human TVRD1 with a KD of less than about 50 nM.

120. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof binds human TRDV with a K _D of less than about 10 nM.

121. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof binds human TRDV with a K _D of less than about 5 nM.

122. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof binds human TRDV with a K _D of less than about 1 nM.

123. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen-binding fragment thereof binds cynomolgus monkey TRDV1 with a K _D of less than about 100 nM.

124. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen-binding fragment thereof binds cynomolgus monkey TRDV1 with a K _D of less than about 50 nM.

125. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen binding fragment thereof downregulates a TCR with an IC50 of less than about 50nM, less than about 10nM, less than about 5nM, less than about 1nM or less than about 0.5nM.

126. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen-binding fragment thereof has an IC90 for TCR downregulation of less than about 10nM or less than about 5nM

127. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen binding fragment thereof kills THP-1 cells with an IC50 of less than about 10nM, less than about 5nM or less than about 1nM.

128. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the anti-vδ1 antibody or antigen binding fragment thereof kills THP-1 cells with an IC90 of less than about 50nM, less than about 10nM, less than about 5nM or less than about 1nM.

129. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. Binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

b. Optionally binds to cynomolgus monkey TRDV1 (SEQ ID NO 308) with a binding affinity (K _D, e.g. measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and

E. the IC90 for killing THP-1 cells is less than about 50nM.

130. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

b. binds to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

c. The IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM) and the IC90 for TCR downregulation is less than about 100nM (less than about 50 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and

E. the IC90 for killing THP-1 cells is less than about 50nM.

131. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM, preferably less than about 5nM,

B. binds to cynomolgus monkey TRDV1 with a K _D of less than about 100nM (preferably less than about 50 nM);

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and

E. the IC90 for killing THP-1 cells is less than about 50nM.

132. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. binds to human TRDV1 with a K _D of less than about 10nM, preferably less than about 1nM,

B. Binding to cynomolgus monkey TRDV1 with a K _D of less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 1nM (preferably less than about 0.5 nM) and the IC90 for TCR downregulation is less than about 10nM (less than about 5 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and

E. the IC90 for killing THP-1 cells is less than about 50nM.

133. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

c. an IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM); and

D. The IC90 for killing THP-1 cells is less than about 50nM.

134. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said anti-vδ1 antibody or antigen binding fragment thereof:

a. Binds to human TRDV1 with a K _D of less than about 10nM (preferably less than about 5 nM);

b. the IC50 for TCR downregulation is less than about 5nM (preferably less than about 1 nM);

c. the IC90 for TCR downregulation is less than about 10nM (preferably less than about 5 nM);

d. An IC50 for killing THP-1 cells of less than about 5nM (preferably less than about 1 nM); and

E. The IC90 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM).

135. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof is a human antibody or antigen binding fragment thereof.

136. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody is scFv, fab, fab ', F (ab') 2, fv, variable domain (e.g. VH or VL), diabody, minibody or full length antibody.

137. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody is a scFv or full length antibody, e.g. IgG1

138. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof is a IgA, igD, igE, igG, igM or IgY antibody.

139. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof is an IgG antibody.

140. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof is an IgG1 antibody.

141. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody is a monoclonal antibody.

142. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said antibody or antigen-binding fragment thereof comprises a VH and a VL, and said VH region and said VL region are joined by a linker, such as a polypeptide linker.

143. The anti-vδ1 antibody of embodiment 142 or an antigen binding fragment thereof, wherein the linker comprises a (Gly 4 Ser) n format, where n=1 to 8.

144. The anti-vδ1 antibody of embodiment 142, or an antigen binding fragment thereof, wherein the linker comprises a [ (Gly ₄Ser)_n(Gly₃AlaSer)_m]_p linker where n, m and p=1 to 8).

145. The anti-vδ1 antibody of embodiment 142 or an antigen binding fragment thereof, wherein said linker comprises the amino acid sequence of SEQ ID NO:291.

146. The anti-vδ1 antibody of embodiment 142 or an antigen binding fragment thereof, wherein said linker consists of SEQ ID NO: 291.

147. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein the antibody or antigen-binding fragment thereof binds only to an epitope in the V region of the vδ1 chain of a γδ TCR.

148. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof binds only to the amino acid sequence of SEQ ID NO: 272. SEQ ID NO:306 or SEQ ID NO:308 amino acid residues 1-90.

149. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof does not bind to an epitope found in CDR3 of the vδ1 chain of a γδ TCR.

150. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein said antibody or antigen binding fragment thereof does not bind to SEQ ID NO:272 within amino acid region 91-105 (CDR 3).

151. The anti-vδ1 antibody or antigen-binding fragment thereof of any preceding embodiment, wherein said antibody or antigen-binding fragment thereof binds to an epitope within the amino acid region

(I) SEQ ID NO:272 3-20; and/or

(Ii) SEQ ID NO:272 from 37 to 77.

152. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 from amino acid residues 37-53 and/or 59-77.

153. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

154. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof consists of the amino acid sequence of SEQ ID NO:272, amino acid residues 37, 42, 50, 53, 59, 64, 68, 69, 72, 73 and 77.

155. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 from amino acid regions 5-20 and/or 62-77.

156. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof comprises the amino acid sequence of SEQ ID NO:272 amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77.

157. The anti-vδ1 antibody of embodiment 151 or an antigen binding fragment thereof, wherein the epitope bound by said antibody or antigen binding fragment thereof consists of the amino acid sequence of SEQ ID NO:272, amino acid residues 5, 9, 16, 20, 62, 64, 72 and 77.

158. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the epitope is an activating epitope of γδ T cells.

159. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 158, wherein binding of the activating epitope: (i) down-regulating γδ TCR; (ii) activate degranulation of γδ T cells; and/or (iii) promote γδ T cell mediated killing.

160. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 158 or 159, wherein binding of said activating epitope upregulates expression of CD107a, CD25, CD69 and/or Ki 67.

161. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the antibody is a multispecific antibody that specifically binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) and a second antigen, or binds to two different epitopes on a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR).

162. The anti-vδ1 antibody of embodiment 161 or an antigen binding fragment thereof, wherein the second antigen is a cell surface protein.

163. The anti-vδ1 antibody of embodiments 161 or 162, or an antigen binding fragment thereof, wherein said second antigen is on a vδ1+ T cell.

164. The anti-vδ1 antibody of embodiments 161 or 162, or an antigen binding fragment thereof, wherein said second antigen is not on a vδ1 cell.

165. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 161 or 162 or 163, wherein said second antigen is on an immune cell.

166. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 161-165, wherein the second antigen is a cancer antigen or a cancer-associated antigen.

167. The anti-vδ1 antibody or antigen binding fragment thereof of embodiment 166, wherein the cancer antigen or cancer-associated antigen is a tumor-associated antigen (TAA).

168. The anti-vδ1 antibody of embodiment 166, or an antigen binding fragment thereof, wherein the cancer antigen or cancer-associated antigen is selected from the group consisting of: AFP, AKAP-4, ALK, alpha fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCA, bcr-abl, beta-catenin, beta-HCG, beta-human chorionic gonadotrophin, BORIS, BTAA, CA, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, CAM 17.1, CAM43, carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33/IL3Ra, CD68\P1, CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG 4), C-Met, CO-029, CSPG4, cyclin B1, cyclophilin C-related protein, CYP1B1, E2A-PRL, EGFR, EGFRvIII, ELF2M, epCAM, ephA, ephran B2, ephran-Bar virus antigen EBVA, ERG (TMPRSS 2ETS fusion gene) ETV6-AML, FAP, FGF-5, fos associated antigen 1, fucosyl GM1, G250, ga733\EpCAM, GAGE-1, GAG E-2, GD3, glioma associated antigen, globoH, glycolipid F77, GM3, GP100 (Pmel 17), H4-RET, HER-2/Neu/ErbB-2, high molecular weight melanoma associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin Growth Factor (IGF) -I, intestinal carboxylesterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, legumain, LMP2, M344, MA-50, HIV-2, mac-2 binding protein 、MAD-CT-1、MAD-CT-2、MAGE、MAGE A1、MAGE A3、MAGE-1、MAGE-3、MAGE-4、MAGE-5、MAGE-6、MART-1、MART-1/MelanA、M-CSF、 melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin, MG7-Ag, ML-IAP, MN-CA IX, MOV18, MUC1, mum-1, hsp70-2, MYCN, MYL-RAR, NA17, NB/70K, neuronal-glial antigen 2 (NG 2), neutrophil elastase, nm-23H1, nuMa, NY-BR-1, NY-CO-1, NY-ESO-1, OY-TES1, p15, p16, p180erbB3, p185erbB2, p53 mutant, page4, PAX3, PAX5, PDGFR-beta, PLAC1, polysialic acid, POLYSAC prostate cancer tumor antigen-1 (PCTA-1), prostate specific antigen, prostatic Acid Phosphatase (PAP), protease 3 (PR 1), PSA, PSCA, PSMA, RAGE-1, ras mutant, RCAS1, RGS5, rhoC, ROR1, RU2 (AS), SART3, SDCCAG, sLe (a), sperm protein 17, SSX2, STn, survivin, TA-90, TAAL, TAG-72, telomerase, thyroglobulin, tie 2, TLP, tn, TPS, TRP-1, TRP-2, TSP-180, tyrosinase, VEGF, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72.

169. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-168, wherein the multispecific antibody is a T cell engagement bispecific antibody.

170. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-169, wherein said antibody is a bispecific antibody and said second antigen is CD19, her2, EGFR, fapα or MSLN.

171. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-170, wherein the antibody is a bispecific antibody and the second antigen is CD19.

172. The anti-vδ1 antibody of embodiment 171 or an antigen binding fragment thereof, wherein said antibody comprises a sequence selected from the group consisting of: SEQ ID NO: 314. SEQ ID NO: 315. SEQ ID NO: 316. SEQ ID NO:396 and SEQ ID NO: 397. SEQ ID NO:421 and SEQ ID NO: 422. SEQ ID NO:504 and SEQ ID NO: 422. SEQ ID NO:423 and SEQ ID NO:424.

173. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-170, wherein said antibody is a bispecific antibody and said second antigen is Her2 (CD 340).

174. The anti-vδ1 antibody of embodiment 173, or an antigen binding fragment thereof, wherein the antibody comprises a sequence selected from the group consisting of: SEQ ID NO: 393. SEQ ID NO: 394. SEQ ID NO: 395. SEQ ID NO: 398. SEQ ID NO:414 and SEQ ID NO:433, respectively; and SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:433, respectively; and SEQ ID NO:505 and SEQ ID NO:434.

175. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-170, wherein the antibody is a bispecific antibody and the second antigen is EGFR.

176. The anti-vδ1 antibody of embodiment 175 or an antigen binding fragment thereof, wherein said antibody comprises SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：385;SEQ ID NO：386;SEQ ID NO：387;SEQ ID NO：388;SEQ ID NO：399;SEQ ID NO：425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:414 and SEQ ID NO:435 or SEQ ID NO:414 and SEQ ID NO:435.

177. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-170, wherein the antibody is a bispecific antibody and the second antigen is fapα.

178. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 177, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:458, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:459 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:460, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:46l, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:462 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:463, a VLCDR3 of the amino acid sequence of seq id no;

179. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 178, said anti-vδ1 antibody or antigen-binding fragment thereof further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

180. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 177, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:456 or a VH consisting of said amino acid sequence and a nucleotide sequence comprising SEQ ID NO:457 or a VL consisting of said amino acid sequence.

181. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 180, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

182. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 177 to 181, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:402.

183. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 177-182, wherein said antibody comprises the amino acid sequence of SEQ ID NO:401; or SEQ ID NO:414 and SEQ ID NO:415.

184. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-170, wherein said antibody is a bispecific antibody and said second antigen is MSLN.

185. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 184, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:466, comprising the amino acid sequence of SEQ ID NO:467 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: VHCDR3 of the amino acid sequence of 468; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:469, comprising the amino acid sequence of SEQ ID NO:470 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:471, a VLCDR3 of the amino acid sequence of seq id no;

186. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 185, further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

187. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 184, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:464 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:465 or a VL consisting of said amino acid sequence.

188. An anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 187, comprising an amino acid sequence comprising SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

189. The anti-vδl antibody or antigen-binding fragment thereof of any one of embodiments 184-188, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:404.

190. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 184-189, wherein said antibody comprises the amino acid sequence of SEQ ID NO:403; or SEQ ID NO:414 and SEQ ID NO:416.

191. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 161-165, wherein the second antigen is an immunomodulatory antigen.

192. The anti-vδ1 antibody of embodiment 191 or an antigen binding fragment thereof, wherein said immunomodulatory antigen may be selected from the group consisting of ：B7-1(CD80)、B7-2(CD86)、B7-DC(CD273)、B7-H1(CD274)、B7-H2(CD275)、B7-H3(CD276)、B7-H4(VTCN1)、B7-H5(VISTA)、BTLA(CD272)、4-1BB(CD137)、CD137L、CD24、CD27、CD28、CD38、CD40、CD40L(CD154)、CD54、CD59、CD70、CTLA4(CD152)、CXCL9、GITR(CD357)、HVEM(CD270)、ICAM-1(CD54)、ICOS(CD278)、LAG-3(CD223)、OX40(CD 134)、OX40L(CD252)、PD-1(CD279)、PD-L1(CD274)、TIGIT、CD314、CD334、CD335、CD337 and TIM-3 (CD 366).

193. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 191 or 192, wherein said second antigen is a stimulatory immune checkpoint molecule

194. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 191 or 192, wherein the second antigen is an inhibitory immune checkpoint molecule.

195. The anti-vδ1 antibody of embodiments 191 or 192, or an antigen binding fragment thereof, wherein said antibody is a bispecific antibody and said second epitope is an epitope of PD-1, 4-1BB (CD 137), OX40 or TIGIT.

196. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 191 or 192, wherein said antibody is a bispecific antibody and said second epitope is an epitope of PD-1.

197. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 196, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:474, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:475 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:476, a VHCDR3 of the amino acid sequence of seq id no; and

B. a light chain variable region comprising a light chain sequence comprising SEQ ID NO:477, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:478 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:479, VLCDR3 of the amino acid sequence;

198. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 197, further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

199. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 195, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:472 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:473 or a VL consisting of said amino acid sequence.

200. The anti-vδ1 antibody of embodiment 199, or an antigen-binding fragment thereof, comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

201. The anti-vδl antibody or antigen-binding fragment thereof of any one of embodiments 195 to 200, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:406.

202. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 195-201, wherein said antibody comprises the amino acid sequence of SEQ ID NO:405; or SEQ ID NO:438 and SEQ ID NO:417.

203. The anti-vδ1 antibody of embodiment 191 or 192, or an antigen binding fragment thereof, wherein said antibody is a bispecific antibody and said second epitope is an epitope of 4-1BB (CD 137).

204. The anti-V.delta.1 antibody of embodiment 203 or an antigen-binding fragment thereof, comprising

A. a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:482, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:483 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 484; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:485, comprising the amino acid sequence of SEQ ID NO:486 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:487 VLCDR3 of the amino acid sequence;

205. The anti-vδl antibody or antigen-binding fragment thereof of embodiment 204, said anti-vδ1 antibody or antigen-binding fragment thereof further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

206. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 203, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:480 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:481 or a VL consisting of said amino acid sequence.

207. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 206, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

208. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 203 to 207, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:408.

209. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 203 to 208, wherein said antibody comprises the amino acid sequence of SEQ ID NO:407, a step of selecting a specific code; or SEQ ID NO:414 and SEQ ID NO:418.

210. The anti-vδ1 antibody or antigen binding fragment thereof of embodiments 191 or 192, wherein the antibody is a bispecific antibody and the second epitope is an epitope of OX 40.

211. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 210, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. a heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:490, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:491 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:492, a VHCDR3 of an amino acid sequence; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:493, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:494 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:495, a VLCDR3 of the amino acid sequence;

212. the anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 211, said anti-vδ1 antibody or antigen-binding fragment thereof further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

213. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 210, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:488 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:489 or a VL consisting of said amino acid sequence.

214. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 213, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

215. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 210 to 214, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:410.

216. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 210-215, wherein said antibody comprises the amino acid sequence of SEQ ID NO:409; or SEQ ID NO:414 and SEQ ID NO:419.

217. The anti-vδ1 antibody of embodiment 191 or 192 or an antigen binding fragment thereof, wherein the antibody is a bispecific antibody and the second epitope is an epitope of TIGIT.

218. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 217, said anti-vδ1 antibody or antigen-binding fragment thereof comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:498, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:499 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:500, a VHCDR3 of an amino acid sequence; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:501, a VLCDR1 comprising the amino acid sequence of SEQ ID NO:502 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:503, a VLCDR3 of the amino acid sequence of seq id no;

219. the anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 218, further comprising

A. A heavy chain variable region comprising a heavy chain sequence comprising SEQ ID NO:51, a VHCDR1 comprising the amino acid sequence of SEQ ID NO:53 and a VHCDR2 comprising the amino acid sequence of SEQ ID NO:68, a VHCDR3 of the amino acid sequence of seq id no; and

B. A light chain variable region comprising a light chain sequence comprising SEQ ID NO:79, comprising the amino acid sequence of SEQ ID NO:80 and a VLCDR2 comprising the amino acid sequence of SEQ ID NO:95, a VLCDR3 of the amino acid sequence of seq id no;

220. An anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 217, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising SEQ ID NO:496 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:497 or a VL consisting of said amino acid sequence.

221. The anti-vδ1 antibody or antigen-binding fragment thereof of embodiment 220, said anti-vδ1 antibody or antigen-binding fragment thereof comprising a polypeptide comprising the amino acid sequence of SEQ ID NO:15 or a VH consisting of said amino acid sequence and a polypeptide comprising SEQ ID NO:40 or a VL consisting of said amino acid sequence.

222. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 217 to 221, said anti-vδ1 antibody or antigen-binding fragment thereof comprising the amino acid sequence of SEQ ID NO:412.

223. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 217 to 222, wherein said antibody comprises the amino acid sequence of SEQ ID NO:411; or SEQ ID NO:439 and SEQ ID NO:420.

224. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 161-223, wherein the second antigen is a cluster of differentiation CD antigen.

225. The anti-vδ1 antibody of embodiment 224 or an antigen binding fragment thereof, wherein said CD antigen is selected from the group consisting of ：CD1a、CD1b、CD1c、CD1d、CD1e、CD2、CD3、CD3d、CD3e、CD3g、CD4、CD5、CD6、CD7、CD8、CD8a、CD8b、CD9、CD10、CD11a、CD11b、CD11 c、CD11d、CD13、CD14、CD15、CD16、CD16a、CD16b、CD17、CD18、CD19、CD20、CD21、CD22、CD23、CD24、CD25、CD26、CD27、CD28、CD29、CD30、CD31、CD32A、CD32B、CD33、CD34、CD35、CD36、CD37、CD38、CD39、CD40、CD41、CD42、CD42a、CD42b、CD42c、CD42d、CD43、CD44、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD50、CD51、CD52、CD53、CD54、CD55、CD56、CD57、CD58、CD59、CD60a、CD60b、CD60c、CD61、CD62E、CD62L、CD62P、CD63、CD64a、CD65、CD65s、CD66a、CD66b、CD66c、CD66d、CD66e、CD66f、CD68、CD69、CD70、CD71、CD72、CD73、CD74、CD75、CD75s、CD77、CD79A、CD79B、CD80、CD81、CD82、CD83、CD84、CD85A、CD85B、CD85C、CD85D、CD85F、CD85G、CD85H、CD85I、CD85J、CD85K、CD85M、CD86、CD87、CD88、CD89、CD90、CD91、CD92、CD93、CD94、CD95、CD96、CD97、CD98、CD99、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107、CD107a、CD107b、CD108、CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120、CD120a、CD120b、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140A、CD140B、CD141、CD142、CD143、CD144、CDw145、CD146、CD147、CD148、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD157、CD158、CD158A、CD158B1、CD158B2、CD158C、CD158D、CD158E1、CD158E2、CD158F1、CD158F2、CD158G、CD158H、CD158I、CD158J、CD158K、CD159a、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CDw198、CDw199、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218a、CD218b、CD219、CD220、CD221、CD222、CD223、CD224、CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240CE、CD240D、CD241、CD242、CD243、CD244、CD245、CD246、CD247、CD248、CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261、CD262、CD263、CD264、CD265、CD266、CD267、CD268、CD269、CD270、CD271、CD272、CD273、CD274、CD275、CD276、CD277、CD278、CD279、CD280、CD281、CD282、CD283、CD284、CD285、CD286、CD287、CD288、CD289、CD290、CD291、CD292、CDw293、CD294、CD295、CD296、CD297、CD298、CD299、CD300A、CD300C、CD301、CD302、CD303、CD304、CD305、CD306、CD307、CD307a、CD307b、CD307c、CD307d、CD307e、CD308、CD309、CD310、CD311、CD312、CD313、CD314、CD315、CD316、CD317、CD318、CD319、CD320、CD321、CD322、CD323、CD324、CD325、CD326、CD327、CD328、CD329、CD330、CD331、CD332、CD333、CD334、CD335、CD336、CD337、CD338、CD339、CD340、CD344、CD349、CD351、CD352、CD353、CD354、CD355、CD357、CD358、CD360、CD361、CD362、CD363、CD364、CD365、CD366、CD367、CD368、CD369、CD370 and CD371.

226. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-225, wherein said antibody is a bispecific antibody.

227. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-226, wherein said antibody is a bispecific antibody and is selected from the group consisting of: cross Mab, DAF (two-in-one), DAF (four-in-one), dutaMab, DT-IgG, pestle-mortar structure (KIH), pestle-mortar structure (universal light chain), charge pair, fab arm exchange, SEEDbody, triomab, LUZ-Y, fcab, kappa lambda body, orthogonal Fab、DVD-IgG、IgG(H)-scFv、scFv-(H)IgG、IgG(L)-scFv、scFv-(L)IgG、IgG(L,H)-Fv、IgG(H)-V、V(H)-IgG、IgG(L)-V、V(L)-IgG、KIH IgG-scFab、2scFv-IgG、IgG-2scFv、scFv4-Ig、Zybody、DVI-IgG( four-in-one), nanobody, nanoby-HAS, biTE, diabody, DART, tandAb, scDiabody, scDiabody-CH3, diabody-CH3, triabody, morisen format, minibody, triBi minibody, scFv-CH3 KIH, fab-scFv, scFv-CH-CL-scFv, F (ab') 2, F (ab) 2-scFv2, scFv-KIH, fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabody-Fc, tandem scFv-Fc, intracellular antibody, dock and Lock, immTAC, HSAbody, scDiabody-HAS, tandem scFv-toxin, igG-ovX-B0 dy, duobody, mab ² and 1-PEG-2.

228. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-227, wherein said antibody is a bispecific antibody conjugate (e.g. an IgG2, scFv, tandem scFv, F (ab') ₂, kiH bispecific antibody, morrison format bispecific antibody (IgG-HC-scFv), diabody, triabody, minibody, or full length bispecific antibody.

229. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-225, wherein said antibody is not a bispecific antibody.

230. The anti-vδ1 antibody or antigen binding fragment thereof of any preceding embodiment, wherein the multispecific antibody is greater than about 70KDa.

231. The anti-vδ1 antibody or antigen binding fragment thereof of any one of embodiments 161-230, wherein said multispecific antibody comprises a sequence ：SEQ ID NO：385、SEQ ID NO：386、SEQ ID NO：387、SEQ ID NO：314、、SEQ ID、SEQ ID NO：394、SEQ ID NO：395,SEQ ID NO：397、SEQ ID NO：402、SEQ ID NO：404、SEQ ID NO：406、SEQ ID NO：408、SEQ ID NO：410、SEQ ID NO：412 selected from the group consisting of SEQ ID NO:413.

232. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of embodiments 161-230, wherein said multispecific antibody comprises SEQ ID NO：388;SEQ ID NO：315;SEQ ID NO：316;SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：393;SEQ ID NO：396;SEQ ID NO：398;SEQ ID NO：399;SEQ ID NO：401;SEQ ID NO：403;SEQ ID NO：405;SEQ ID NO：407;SEQ ID NO：409;SEQ ID NO：411;SEQ ID NO：414 and SEQ ID NO:415, a base; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:418; SEQ ID NO:505 and SEQ ID NO:415, a base; SEQ ID NO:505 and SEQ ID NO:416; SEQ ID NO:505 and SEQ ID NO:419; SEQ ID NO:505 and SEQ ID NO:418; SEQ ID NO:421 and SEQ ID NO:422, a part of the material; SEQ ID NO:504 and SEQ ID NO:422, a part of the material; SEQ ID NO:423 and SEQ ID NO: 424. SEQ ID NO:425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:429; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:414 and SEQ ID NO:435; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:434, a base; SEQ ID NO:505 and SEQ ID NO:435; SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

233. A polynucleotide sequence encoding an anti-vδ1 antibody or antigen binding fragment thereof as defined in any preceding embodiment.

234. A polynucleotide sequence encoding a polypeptide comprising a sequence identical to SEQ ID NO:199 to 222, 224 to 247, 249 to 259, or 261 to 271, or an antibody binding fragment thereof.

235. An expression vector comprising a polynucleotide sequence as defined in embodiment 233 or embodiment 234.

236. A host cell comprising a polynucleotide sequence as defined in embodiment 233 or embodiment 234 or an expression vector as defined in embodiment 235, optionally wherein the host cell is a recombinant host cell.

237. The host cell of embodiment 236, comprising a first polynucleotide sequence encoding a VH sequence and a second polynucleotide sequence encoding a VL sequence.

238. The host cell of embodiment 236, comprising a first expression vector comprising a first polynucleotide sequence encoding a VH sequence and a second expression vector comprising a second polynucleotide sequence encoding a VL sequence.

239. The host cell of embodiment 236, comprising an expression vector comprising a first polynucleotide sequence encoding a VH sequence and a second polynucleotide sequence encoding a VL sequence.

240. A host cell as defined in any one of embodiments 236 to 239, wherein the polynucleotide or expression vector encodes a membrane anchor or transmembrane domain fused to the antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is presented on the cell's outer surface

241. A method for producing any one of the antibodies or antigen-binding fragments thereof of any one of embodiments 1 to 232, the method comprising culturing the host cell of any one of embodiments 236 to 240 in a cell culture medium.

242. The method of embodiment 241, further comprising isolating said antibody or antigen-binding fragment thereof from said culture medium.

243. A method of mutating an antibody or antigen binding fragment thereof, the method comprising providing an antibody comprising a kappa light chain having serine at position 74 according to the IMGT numbering system, and mutating the serine to a different amino acid.

244. The method of embodiment 243, wherein the antibody is an anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof.

245. The method of embodiment 243 or embodiment 244, wherein the antibody is of a sequence comprising a sequence identical to SEQ ID NO:1 and VH comprising an amino acid sequence at least 90% identical to SEQ ID NO:26, an antibody to VL of an amino acid sequence that is at least 90% identical.

246. The method of any one of embodiments 243 to 244, wherein said antibody or antigen-binding fragment thereof binds a human antigen and said mutation increases the affinity of said antibody or antigen-binding fragment thereof for a cognate cynomolgus monkey antigen.

247. The method of any one of embodiments 243 to 246, wherein the antibody or antigen binding fragment thereof after and after the mutation is introduced is an anti-tcrδ variable 1 (anti-vδ1) antibody or antigen binding fragment thereof that binds to a variable δ1 (vδ1) chain of a human γδ T Cell Receptor (TCR), and the affinity of the antibody for a variable δ1 (vδ1) chain of a cynomolgus monkey γδ T Cell Receptor (TCR) is increased after the mutation is introduced.

248. A method of making a variant anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising providing a parent antibody comprising:

a. Comprising SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, VL of the amino acid sequence of seq id no;

b. comprising SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, VL of the amino acid sequence of seq id no;

c. comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

d. Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

e. comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

f. comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

g. comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

h. Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

i. comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

j. comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

k. Comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

L. comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, VL of the amino acid sequence of seq id no;

And subjecting the antibody to affinity maturation, wherein the produced antibody has a greater affinity for the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) than the parent antibody.

249. The method of embodiment 248, wherein the produced antibody binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) with an affinity that is at least 20%, at least 30%, at least 40%, more preferably at least 50%, at least 100% or more preferably at least about 1000% greater than the parent antibody, e.g. as measured by Kd.

250. The method of embodiment 248 or embodiment 249, further comprising co-formulating the produced antibodies with at least one or more pharmaceutically acceptable diluents or carriers.

251. A method of preparing a pharmaceutical composition comprising providing an antibody prepared according to the method of any one of embodiments 241 to 250 and co-formulating the antibody with at least one or more pharmaceutically acceptable diluents or carriers.

252. An anti-vδ1 antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof being prepared according to or being prepared according to the method of any one of embodiments 241 to 250.

253. A pharmaceutical composition prepared according to or preparable according to the method of embodiment 251.

254. A composition comprising an antibody or antibody binding fragment thereof as defined in any one of embodiments 1 to 23l or 251.

255. A pharmaceutical composition comprising an antibody or antibody-binding fragment thereof as defined in any one of embodiments 1 to 231 or 251 and a pharmaceutically acceptable diluent or carrier.

256. A kit comprising the anti-vδ1 antibody or antibody binding fragment of any one of embodiments 1 to 229 or 249 or the pharmaceutical composition of embodiments 253 or 255.

257. The kit of embodiment 256, further comprising an additional therapeutically active agent.

258. The kit of embodiment 256 or embodiment 257, further comprising instructions for use.

259. A method of treating a disease or disorder in a subject, the method comprising administering to the subject the anti-vδ1 antibody or antibody binding fragment thereof of any one of embodiments 1 to 231 or 251, or the pharmaceutical composition of embodiments 253 or 255.

260. The method of embodiment 259, wherein the disease or disorder is cancer, infectious disease, or inflammatory disease.

261. The method of any one of embodiments 259 to 260, further comprising administering a second agent to the subject simultaneously or sequentially in any order.

262. A method of modulating an immune response in a subject, the method comprising administering to the subject the anti-vδ1 antibody or antibody binding fragment thereof of any one of embodiments 1 to 231 or 251, or the pharmaceutical composition of embodiments 253 or 255.

263. The method of embodiment 262, wherein the subject has cancer, infectious disease, or inflammatory disease.

264. The method of embodiment 262 or 263, wherein modulating the immune response in the subject comprises at least one selected from the group consisting of: activating γδ T cells, causing or increasing γδ T cell proliferation, causing or increasing γδ T cell expansion, causing or increasing γδ T cell degranulation, causing or increasing γδ T cell mediated killing activity, causing or increasing γδ T cytotoxicity, causing or increasing γδ T cell mobilization, increasing γδ T cell survival, and increasing resistance to γδ T cell depletion.

265. The method of any one of embodiments 259 to 264, wherein the diseased cells are killed and healthy cells are retained.

266. The anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, of any one of embodiments 1-231 or 251, or the pharmaceutical composition of embodiment 253 or 25, or the kit of any one of embodiments 256-258, for use in medicine.

267. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 1-231 or 251, or the pharmaceutical composition of embodiment 253 or 255, or the kit of any one of embodiments 256-258, for use in the treatment of cancer, infectious disease, or inflammatory disease.

268. Use of the anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 1-231 or 251 for the manufacture of a medicament.

269. Use of the anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 1-231 or 251 for the manufacture of a medicament for the treatment of cancer, infectious disease, or inflammatory disease.

270. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of embodiments 103-110, wherein the antibody is an antibody as defined in any one of embodiments 1-101.

Sequence listing

<110> Aldata biopharmaceutical Co., ltd (Adaptate Biotherapeutics Ltd)

<120> Anti-TCR delta variable 1 antibodies

<130> P142024WO

<150> GB2102224.9

<151> 2021-02-17

<150> GB2111685.0

<151> 2021-08-14

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50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Ser Trp Val Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 17

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 variable heavy chain

<400> 17

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Ser Trp Val Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 18

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 variable heavy chain

<400> 18

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Ser Trp Val Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 19

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 variable heavy chain

<400> 19

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ala Asp Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 20

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 variable heavy chain

<400> 20

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Ala Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 21

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 variable heavy chain

<400> 21

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Val Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Leu Gly Asn Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 22

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 variable heavy chain

<400> 22

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Ala Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 23

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 variable heavy chain

<400> 23

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Ala Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 24

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 variable heavy chain

<400> 24

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ala Gly Tyr Pro Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 25

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 variable heavy chain

<400> 25

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ala Asp Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 26

<211> 110

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 variable light chain

<400> 26

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

20 25 30

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105 110

<210> 27

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-105 variable light chain

<400> 27

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Gln

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 28

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-107 variable light chain

<400> 28

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Pro Gln

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 29

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 variable light chain

<400> 29

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Gln

85 90 95

Val Thr Phe Gly Ala Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 30

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-112 variable light chain

<400> 30

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Gln Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 31

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-117 variable light chain

<400> 31

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Pro Arg

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 32

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-19 variable light chain

<400> 32

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Lys

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 33

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21 variable light chain

<400> 33

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Trp

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 34

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-31 variable light chain

<400> 34

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Pro Pro

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 35

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-139 variable light chain

<400> 35

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Ser Thr Pro Gln

85 90 95

Leu Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 36

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-4 variable light chain

<400> 36

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Trp

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 37

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 variable light chain

<400> 37

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr His Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Val Ile Lys

100 105

<210> 38

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-53 variable light chain

<400> 38

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Gln

85 90 95

Leu Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 39

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-173 variable light chain

<400> 39

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 40

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 variable light chain

<400> 40

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 41

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-8 variable light chain

<400> 41

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 42

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 variable light chain

<400> 42

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Pro Gln

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 43

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 variable light chain

<400> 43

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Glu Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 44

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 variable light chain

<400> 44

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Glu

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 45

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 variable light chain

<400> 45

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Asp Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 46

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 variable light chain

<400> 46

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Glu Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 47

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 variable light chain

<400> 47

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Thr Pro Gln

85 90 95

Val Thr Phe Gly Ser Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 48

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 variable light chain

<400> 48

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Pro Ile

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 49

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 variable light chain

<400> 49

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Lys Thr Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 50

<211> 108

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 variable light chain

<400> 50

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Tyr Ser Thr Asp Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 51

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VHCDR1 (parent and affinity matured clone)

<400> 51

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala

1 5 10

<210> 52

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21/ADT1-4-53/ADT1-4-86 VHCDR1

<400> 52

Gly Asp Ser Val Ser Ser Lys Ser Val Ala

1 5 10

<210> 53

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VHCDR2 (parent and all affinity matured clones)

<400> 53

Thr Tyr Tyr Arg Ser Lys Trp Ser Thr

1 5

<210> 54

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VHCDR3

<400> 54

Thr Trp Ser Gly Tyr Val Asp Val

1 5

<210> 55

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-105 VHCDR3

<400> 55

Thr Trp Val Gly Tyr Val Asp Val

1 5

<210> 56

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-107 VHCDR3

<400> 56

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 57

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 VHCDR3

<400> 57

Thr Trp Val Glu Tyr Val Asp Val

1 5

<210> 58

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-112 VHCDR3

<400> 58

Thr Trp Val Gly Tyr Val Asp Tyr

1 5

<210> 59

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-117 VHCDR3

<400> 59

Thr Trp Val Gly Tyr Val Asp Val

1 5

<210> 60

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-19 VHCDR3

<400> 60

Thr Trp Val Gly Tyr Val Asp Arg

1 5

<210> 61

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21 VHCDR3

<400> 61

Thr Trp Ala Asp Tyr Val Asp Val

1 5

<210> 62

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-31 VHCDR3

<400> 62

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 63

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-139 VHCDR3

<400> 63

Thr Trp Val Gly Tyr Val Asp Tyr

1 5

<210> 64

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-4 VHCDR3

<400> 64

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 65

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 VHCDR3

<400> 65

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 66

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-53 VHCDR3

<400> 66

Thr Trp Ala Asp Tyr Val Asp Val

1 5

<210> 67

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-173 VHCDR3

<400> 67

Thr Trp Ala Gly Tyr Pro Asp Val

1 5

<210> 68

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 VHCDR3

<400> 68

Thr Trp Val Gly Tyr Val Asp Arg

1 5

<210> 69

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-8 VHCDR3

<400> 69

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 70

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 VHCDR3

<400> 70

Ser Trp Val Gly Tyr Val Asp Val

1 5

<210> 71

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 VHCDR3

<400> 71

Ser Trp Val Gly Tyr Val Asp Val

1 5

<210> 72

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 VHCDR3

<400> 72

Thr Trp Ala Asp Tyr Val Asp Val

1 5

<210> 73

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 VHCDR3

<400> 73

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 74

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 VHCDR3

<400> 74

Thr Trp Leu Gly Asn Val Asp Val

1 5

<210> 75

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 VHCDR3

<400> 75

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 76

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 VHCDR3

<400> 76

Thr Trp Val Gly Tyr Ala Asp Val

1 5

<210> 77

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 VHCDR3

<400> 77

Thr Trp Ala Gly Tyr Pro Asp Val

1 5

<210> 78

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 VHCDR3

<400> 78

Thr Trp Ala Asp Tyr Val Asp Val

1 5

<210> 79

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VLCDR1 (parent and all affinity matured clones)

<400> 79

Gln Asp Ile Asn Asp Trp

1 5

<210> 80

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VLCDR2 (parent and all affinity matured clones)

<400> 80

Asp Ala Ser

1

<210> 81

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 VLCDR3

<400> 81

Gln Gln Ser Tyr Ser Thr Pro Gln Val Thr

1 5 10

<210> 82

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-105 VLCDR3

<400> 82

Gln Gln Lys Tyr Ser Thr Pro Gln Ile Thr

1 5 10

<210> 83

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-107 VLCDR3

<400> 83

Gln Gln Arg Tyr Ser Thr Pro Gln Ile Thr

1 5 10

<210> 84

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 VLCDR3

<400> 84

Gln Gln Lys Tyr Ser Thr Pro Gln Val Thr

1 5 10

<210> 85

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-112 VLCDR3

<400> 85

Gln Gln Lys Tyr Ser Gln Pro Gln Val Thr

1 5 10

<210> 86

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-117 VLCDR3

<400> 86

Gln Gln Arg Tyr Ser Thr Pro Arg Val Thr

1 5 10

<210> 87

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-19 VLCDR3

<400> 87

Gln Gln Lys Tyr Ser Thr Pro Lys Val Thr

1 5 10

<210> 88

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21 VLCDR3

<400> 88

Gln Gln Lys Tyr Ser Thr Pro Trp Val Thr

1 5 10

<210> 89

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-31 VLCDR3

<400> 89

Gln Gln Arg Tyr Ser Thr Pro Pro Val Thr

1 5 10

<210> 90

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-139 VLCDR3

<400> 90

Gln Gln Gly Tyr Ser Thr Pro Gln Leu Thr

1 5 10

<210> 91

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-4 VLCDR3

<400> 91

Gln Gln Lys Tyr Ser Thr Pro Trp Val Thr

1 5 10

<210> 92

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 VLCDR3

<400> 92

Gln Gln Arg Tyr Ser Thr His Gln Val Thr

1 5 10

<210> 93

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-53 VLCDR3

<400> 93

Gln Gln Lys Tyr Ser Thr Pro Gln Leu Thr

1 5 10

<210> 94

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-173 VLCDR3

<400> 94

Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr

1 5 10

<210> 95

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 VLCDR3

<400> 95

Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr

1 5 10

<210> 96

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-8 VLCDR3

<400> 96

Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr

1 5 10

<210> 97

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 VLCDR3

<400> 97

Gln Gln Arg Tyr Ser Thr Pro Gln Ile Thr

1 5 10

<210> 98

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 VLCDR3

<400> 98

Gln Gln Lys Tyr Ser Glu Pro Gln Val Thr

1 5 10

<210> 99

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 VLCDR3

<400> 99

Gln Gln Lys Tyr Ser Thr Pro Glu Val Thr

1 5 10

<210> 100

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 VLCDR3

<400> 100

Gln Gln Lys Tyr Ser Asp Pro Gln Val Thr

1 5 10

<210> 101

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 VLCDR3

<400> 101

Gln Gln Lys Tyr Ser Glu Pro Gln Val Thr

1 5 10

<210> 102

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 VLCDR3

<400> 102

Gln Gln Lys Tyr Ser Thr Pro Gln Val Thr

1 5 10

<210> 103

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 VLCDR3

<400> 103

Gln Gln Arg Tyr Ser Thr Pro Ile Val Thr

1 5 10

<210> 104

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 VLCDR3

<400> 104

Gln Gln Lys Tyr Lys Thr Pro Gln Val Thr

1 5 10

<210> 105

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 VLCDR3

<400> 105

Gln Gln Arg Tyr Ser Thr Asp Gln Val Thr

1 5 10

<210> 106

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 variable heavy chain

<400> 106

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Ala Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 107

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 variable heavy chain

<400> 107

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Tyr Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 108

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 variable heavy chain

<400> 108

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Tyr Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 109

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 variable heavy chain

<400> 109

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ile Asp Tyr Glu Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 110

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 variable heavy chain

<400> 110

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Ser Tyr Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 111

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 variable heavy chain

<400> 111

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ile Asp Tyr Glu Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 112

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 variable heavy chain

<400> 112

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Gln Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 113

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 variable heavy chain

<400> 113

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Tyr Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 114

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 variable heavy chain

<400> 114

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Asn Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 115

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 variable heavy chain

<400> 115

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Tyr Glu Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 116

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 variable heavy chain

<400> 116

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Ser Tyr Ala Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 117

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 variable heavy chain

<400> 117

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Tyr Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 118

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 variable light chain

<400> 118

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly

20 25 30

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 119

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 variable light chain

<400> 119

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Ala Ser Thr Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 120

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 variable light chain

<400> 120

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ala Asp Thr Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 121

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 variable light chain

<400> 121

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Gly Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 122

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 variable light chain

<400> 122

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Val Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ala Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 123

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 variable light chain

<400> 123

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ala Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 124

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 variable light chain

<400> 124

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Gly Leu Asp

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 125

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 variable light chain

<400> 125

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ala Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 126

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 variable light chain

<400> 126

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Gly Leu Asp

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 127

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 variable light chain

<400> 127

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 128

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 variable light chain

<400> 128

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Gly Leu Asp

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 129

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 variable light chain

<400> 129

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly Thr Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ala Ser Glu Leu Leu

85 90 95

Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

100 105

<210> 130

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VHCDR1 (parent and all affinity matured clones)

<400> 130

Gly Phe Thr Phe Ser Asp Tyr Tyr

1 5

<210> 131

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VHCDR2 (parent and all affinity matured clones)

<400> 131

Ile Ser Ser Ser Gly Ser Thr Ile

1 5

<210> 132

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VHCDR3

<400> 132

Val Asp Tyr Ala Asp Ala Phe Asp Ile

1 5

<210> 133

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 VHCDR3

<400> 133

Asp Asp Tyr Asn Asp Ala Phe Asp Ile

1 5

<210> 134

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 VHCDR3

<400> 134

Asp Asp Tyr Asn Asp Ala Phe Asp Ile

1 5

<210> 135

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 VHCDR3

<400> 135

Ile Asp Tyr Glu Asp Ala Phe Asp Ile

1 5

<210> 136

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 VHCDR3

<400> 136

Val Ser Tyr Asp Asp Ala Phe Asp Ile

1 5

<210> 137

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 VHCDR3

<400> 137

Ile Asp Tyr Glu Asp Ala Phe Asp Ile

1 5

<210> 138

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 VHCDR3

<400> 138

Val Asp Tyr Gln Glu Ala Phe Asp Ile

1 5

<210> 139

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 VHCDR3

<400> 139

Asp Asp Tyr Asn Asp Ala Phe Asp Ile

1 5

<210> 140

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 VHCDR3

<400> 140

Val Asp Tyr Asn Glu Ala Phe Asp Ile

1 5

<210> 141

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 VHCDR3

<400> 141

Asp Asp Tyr Glu Asp Ala Phe Asp Ile

1 5

<210> 142

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 VHCDR3

<400> 142

Val Ser Tyr Ala Glu Ala Phe Asp Ile

1 5

<210> 143

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 VHCDR3

<400> 143

Asp Asp Tyr Asp Asp Ala Phe Asp Ile

1 5

<210> 144

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VLCDR1 (parent and all affinity matured clones)

<400> 144

Gln Ser Ile Gly Thr Tyr

1 5

<210> 145

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VLCDR2 (parent and all affinity matured clones)

<400> 145

Val Ala Ser

1

<210> 146

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 VLCDR3

<400> 146

Gln Gln Ser Tyr Ser Thr Leu Leu Thr

1 5

<210> 147

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 VLCDR3

<400> 147

Gln Gln Thr Ala Ser Thr Leu Leu Thr

1 5

<210> 148

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 VLCDR3

<400> 148

Gln Gln Ser Ala Asp Thr Leu Leu Thr

1 5

<210> 149

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 VLCDR3

<400> 149

Gln Gln Ser Gly Ser Glu Leu Leu Thr

1 5

<210> 150

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 VLCDR3

<400> 150

Gln Gln Ser Ala Ser Glu Leu Leu Thr

1 5

<210> 151

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 VLCDR3

<400> 151

Gln Gln Ser Ala Ser Glu Leu Leu Thr

1 5

<210> 152

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 VLCDR3

<400> 152

Gln Gln Ser Tyr Ser Gly Leu Asp Thr

1 5

<210> 153

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 VLCDR3

<400> 153

Gln Gln Ser Ala Ser Glu Leu Leu Thr

1 5

<210> 154

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 VLCDR3

<400> 154

Gln Gln Ser Tyr Ser Gly Leu Asp Thr

1 5

<210> 155

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 VLCDR3

<400> 155

Gln Gln Ser Ser Ser Glu Leu Leu Thr

1 5

<210> 156

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 VLCDR3

<400> 156

Gln Gln Ser Tyr Ser Gly Leu Asp Thr

1 5

<210> 157

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 VLCDR3

<400> 157

Gln Gln Ser Ala Ser Glu Leu Leu Thr

1 5

<210> 158

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR1

<220>

<221> misc_feature

<223> X is any amino acid

<220>

<221> misc_feature

<222> (9)..(9)

<223> Xaa can be any naturally occurring amino acid

<400> 158

Gly Asp Ser Val Ser Ser Lys Ser Xaa Ala

1 5 10

<210> 159

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR1

<220>

<221> misc_feature

<223> X is A or V

<220>

<221> misc_feature

<222> (9)..(9)

<223> Xaa can be any naturally occurring amino acid

<400> 159

Gly Asp Ser Val Ser Ser Lys Ser Xaa Ala

1 5 10

<210> 160

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR1

<220>

<221> misc_feature

<223> X is A or V

<220>

<221> misc_feature

<222> (9)..(9)

<223> Xaa can be any naturally occurring amino acid

<400> 160

Gly Asp Ser Val Ser Ser Lys Ser Xaa Ala

1 5 10

<210> 161

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR1

<220>

<221> misc_feature

<223> X is A or V

<220>

<221> misc_feature

<222> (9)..(9)

<223> Xaa can be any naturally occurring amino acid

<400> 161

Gly Asp Ser Val Ser Ser Lys Ser Xaa Ala

1 5 10

<210> 162

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<223> X is any amino acid

<220>

<221> misc_feature

<222> (1)..(1)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> misc_feature

<222> (3)..(6)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> misc_feature

<222> (8)..(8)

<223> Xaa can be any naturally occurring amino acid

<400> 162

Xaa Trp Xaa Xaa Xaa Xaa Asp Xaa

1 5

<210> 163

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<222> (1)..(1)

<223> X is S or T

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is V, A or L

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is G, E or D

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is Y or N

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is V, A or P

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is V, Y or R

<400> 163

Xaa Trp Xaa Xaa Xaa Xaa Asp Xaa

1 5

<210> 164

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<222> (1)..(1)

<223> X is S or T

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is V, A or L

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is G, E or D

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is Y or N

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is V, A or P

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is V, Y or R

<400> 164

Xaa Trp Xaa Xaa Xaa Xaa Asp Xaa

1 5

<210> 165

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<222> (1)..(1)

<223> X is S or T

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is V, A or L

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is G or D

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is Y or N

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is V, A or P

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is V, Y or R

<400> 165

Xaa Trp Xaa Xaa Xaa Xaa Asp Xaa

1 5

<210> 166

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<223> X is any amino acid

<220>

<221> misc_feature

<222> (3)..(3)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> misc_feature

<222> (5)..(9)

<223> Xaa can be any naturally occurring amino acid

<400> 166

Gln Gln Xaa Tyr Xaa Xaa Xaa Xaa Xaa Thr

1 5 10

<210> 167

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is K, R or G

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is S or K

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is T, Q, A, E or D

<220>

<221> misc_feature

<222> (7)..(7)

<223> X is P, H or D

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is Q, R, K, W, P, E or I

<220>

<221> misc_feature

<222> (9)..(9)

<223> X is I, V or L

<400> 167

Gln Gln Xaa Tyr Xaa Xaa Xaa Xaa Xaa Thr

1 5 10

<210> 168

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is K, R or G

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is S or K

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is T, Q, A, E or D

<220>

<221> misc_feature

<222> (7)..(7)

<223> X is P or H

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is Q, R, K, W, P, E or I

<220>

<221> misc_feature

<222> (9)..(9)

<223> X is I, V or L

<400> 168

Gln Gln Xaa Tyr Xaa Xaa Xaa Xaa Xaa Thr

1 5 10

<210> 169

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is K or R

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is S or K

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is T, Q, A or E

<220>

<221> misc_feature

<222> (7)..(7)

<223> X is P or H

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is Q, K, W, P or I

<220>

<221> misc_feature

<222> (9)..(9)

<223> X is V or L

<400> 169

Gln Gln Xaa Tyr Xaa Xaa Xaa Xaa Xaa Thr

1 5 10

<210> 170

<211> 25

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 HFR1 (parent and affinity maturing clone)

<400> 170

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser

20 25

<210> 171

<211> 25

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 HFR1

<400> 171

Glu Val Gln Leu Leu Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser

20 25

<210> 172

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 HFR2 (parent and all affinity matured clones)

<400> 172

Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly

1 5 10 15

Arg

<210> 173

<211> 40

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 HFR3 (parent and all affinity matured clones)

<400> 173

Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr

1 5 10 15

Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp

20 25 30

Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 174

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 HFR4 (parent and all affinity matured clones)

<400> 174

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 175

<211> 26

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 LFR1 (parental and all affinity matured clones)

<400> 175

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

20 25

<210> 176

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 LFR2 (parental and all affinity matured clones)

<400> 176

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

1 5 10 15

Tyr

<210> 177

<211> 36

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 LFR3 (affinity matured clone)

<400> 177

Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly Ser Gly Ser Gly

1 5 10 15

Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala

20 25 30

Thr Tyr Tyr Cys

35

<210> 178

<211> 36

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 parental clone and ADT1-4-138 LFR3

<400> 178

Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

1 5 10 15

Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala

20 25 30

Thr Tyr Tyr Cys

35

<210> 179

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 LFR4 (parental and affinity matured clone)

<400> 179

Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

1 5 10

<210> 180

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 LFR4

<400> 180

Phe Gly Ala Gly Thr Arg Leu Glu Ile Lys

1 5 10

<210> 181

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 LFR4

<400> 181

Phe Gly Gln Gly Thr Arg Leu Val Ile Lys

1 5 10

<210> 182

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 LFR4

<400> 182

Phe Gly Ser Gly Thr Arg Leu Glu Ile Lys

1 5 10

<210> 183

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<223> X is any amino acid

<220>

<221> misc_feature

<222> (1)..(2)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> misc_feature

<222> (4)..(5)

<223> Xaa can be any naturally occurring amino acid

<400> 183

Xaa Xaa Tyr Xaa Xaa Ala Phe Asp Ile

1 5

<210> 184

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<222> (1)..(1)

<223> X is D, I or V

<220>

<221> misc_feature

<222> (2)..(2)

<223> X is D or S

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is N, E, D, Q or A

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is D or E

<400> 184

Xaa Xaa Tyr Xaa Xaa Ala Phe Asp Ile

1 5

<210> 185

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: HCDR3

<220>

<221> misc_feature

<222> (1)..(1)

<223> X is D or V

<220>

<221> misc_feature

<222> (2)..(2)

<223> X is D or S

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is D, Q or A

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is D or E

<400> 185

Xaa Xaa Tyr Xaa Xaa Ala Phe Asp Ile

1 5

<210> 186

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<223> X is any amino acid

<220>

<221> misc_feature

<222> (3)..(6)

<223> Xaa can be any naturally occurring amino acid

<220>

<221> misc_feature

<222> (8)..(8)

<223> Xaa can be any naturally occurring amino acid

<400> 186

Gln Gln Xaa Xaa Xaa Xaa Leu Xaa Thr

1 5

<210> 187

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is T or S

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is A, G, Y or S

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is S or D

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is T, E or G

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is L or D

<400> 187

Gln Gln Xaa Xaa Xaa Xaa Leu Xaa Thr

1 5

<210> 188

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 derived anti-TCR delta variable 1 antibody or antigen-binding fragment thereof: LCDR3

<220>

<221> misc_feature

<222> (3)..(3)

<223> X is S

<220>

<221> misc_feature

<222> (4)..(4)

<223> X is A or Y

<220>

<221> misc_feature

<222> (5)..(5)

<223> X is S

<220>

<221> misc_feature

<222> (6)..(6)

<223> X is E or G

<220>

<221> misc_feature

<222> (8)..(8)

<223> X is L or D

<400> 188

Gln Gln Xaa Xaa Xaa Xaa Leu Xaa Thr

1 5

<210> 189

<211> 25

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 HFR1 (parent and all affinity matured clones)

<400> 189

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 190

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 HFR2 (parent and all affinity matured clones)

<400> 190

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10 15

Tyr

<210> 191

<211> 40

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 HFR3 (parent and all affinity matured clones)

<400> 191

Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

1 5 10 15

Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

20 25 30

Thr Ala Val Tyr Tyr Cys Ala Arg

35 40

<210> 192

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 HFR4 (parent and all affinity matured clones)

<400> 192

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 193

<211> 26

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 LFR1 (parental and affinity matured clone)

<400> 193

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Ala Gly

20 25

<210> 194

<211> 26

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 LFR1

<400> 194

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Ala Cys Arg Val Gly

20 25

<210> 195

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 LFR2 (parental and all affinity matured clones)

<400> 195

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

1 5 10 15

Tyr

<210> 196

<211> 36

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 LFR3 (parental and all affinity matured clones)

<400> 196

Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

1 5 10 15

Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala

20 25 30

Thr Tyr Tyr Cys

35

<210> 197

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 LFR4 (parental and all affinity matured clones)

<400> 197

Phe Gly Arg Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 198

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 variable heavy chain DNA

<400> 198

caggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtgga gtggttatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 199

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-105 variable heavy chain DNA

<400> 199

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 200

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-107 variable heavy chain DNA

<400> 200

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 201

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 variable heavy chain DNA

<400> 201

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcga tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctctc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tggagtatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 202

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-112 variable heavy chain DNA

<400> 202

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggactactgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 203

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-117 variable heavy chain DNA

<400> 203

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 204

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-19 variable heavy chain DNA

<400> 204

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggacaggtgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 205

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21 variable heavy chain DNA

<400> 205

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ttgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccgactatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 206

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-31 variable heavy chain DNA

<400> 206

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 207

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-139 variable heavy chain DNA

<400> 207

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggactactgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 208

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-4 variable heavy chain DNA

<400> 208

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 209

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 variable heavy chain DNA

<400> 209

gaggtacagc tgctgcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 210

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-53 variable heavy chain DNA

<400> 210

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ttgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccgactatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 211

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-173 variable heavy chain DNA

<400> 211

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccggttatcc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 212

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 variable heavy chain DNA

<400> 212

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgt ggacaggtgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 213

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-8 variable heavy chain DNA

<400> 213

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 214

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 variable heavy chain DNA

<400> 214

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaagctggg tgggttatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 215

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 variable heavy chain DNA

<400> 215

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaagctggg tgggttatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 216

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 variable heavy chain DNA

<400> 216

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccgactatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 217

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 variable heavy chain DNA

<400> 217

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 218

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 variable heavy chain DNA

<400> 218

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ttgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggc tgggtaacgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 219

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 variable heavy chain DNA

<400> 219

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 220

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 variable heavy chain DNA

<400> 220

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg tgggttatgc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 221

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 variable heavy chain DNA

<400> 221

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccggttatcc cgacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 222

<211> 360

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 variable heavy chain DNA

<400> 222

gaggtacagc tgcagcagtc aggtccagga ctggtgaagc cctcgcagac cctctcactc 60

acctgtgcca tctccgggga cagtgtctcc agcaaaagtg ctgcttggaa ctggatcagg 120

cagtccccat cgagaggcct tgagtggctg ggaaggacat actacaggtc caagtggtct 180

actgattatg cagcatctgt gaaaagtcga ataaccatca acccagacac atccaagaac 240

cagctctccc tgcagttaaa ctctgtgact cccgaggaca cggctgtgta ttactgtgca 300

agaacgtggg ccgactatgt ggacgtctgg ggccaaggaa ccctggtcac cgtctcgagc 360

<210> 223

<211> 330

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 variable light chain DNA

<400> 223

gctagcgaca tccagatgac ccagtcccct cccgccctgt ctgcatctgt gggagacaga 60

gtcaccatca cttgccgggc cagtcaagat attaatgact ggttggcctg gtatcagcat 120

aaacctggga aagcccctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 180

ccatcaaggt tcagcggcag tggatctggg acagaattca ctctcaccat cagcagcctg 240

cagcctgatg attttgcaac ttactactgt caacagagtt acagtacccc tcaggtcact 300

tttggccagg ggacacgact ggagatcaaa 330

<210> 224

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-105 variable light chain DNA

<400> 224

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctcagat cacttttggc 300

caggggacac gactggagat caaa 324

<210> 225

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-107 variable light chain DNA

<400> 225

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccctcagat cacttttggc 300

caggggacac gactggagat caaa 324

<210> 226

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-110 variable light chain DNA

<400> 226

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctcaggt cacttttggc 300

gccgggacac gactggagat caaa 324

<210> 227

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-112 variable light chain DNA

<400> 227

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtc agcctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 228

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-117 variable light chain DNA

<400> 228

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccctcgcgt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 229

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-19 variable light chain DNA

<400> 229

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctaaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 230

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-21 variable light chain DNA

<400> 230

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta ccccttgggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 231

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-31 variable light chain DNA

<400> 231

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccctcccgt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 232

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-139 variable light chain DNA

<400> 232

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag ggctacagta cccctcagct gacttttggc 300

caggggacac gactggagat caaa 324

<210> 233

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-4 variable light chain DNA

<400> 233

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta ccccttgggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 234

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-143 variable light chain DNA

<400> 234

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccaccaggt cacttttggc 300

caggggacac gactggtgat caaa 324

<210> 235

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-53 variable light chain DNA

<400> 235

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctcagct gacttttggc 300

caggggacac gactggagat caaa 324

<210> 236

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-173 variable light chain DNA

<400> 236

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg cccctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 237

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 variable light chain DNA

<400> 237

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg cccctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 238

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-8 variable light chain DNA

<400> 238

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg cccctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 239

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-82 variable light chain DNA

<400> 239

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccctcagat cacttttggc 300

caggggacac gactggagat caaa 324

<210> 240

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-83 variable light chain DNA

<400> 240

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg agcctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 241

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-3 variable light chain DNA

<400> 241

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctgaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 242

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-84 variable light chain DNA

<400> 242

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg accctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 243

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-86 variable light chain DNA

<400> 243

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagtg agcctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 244

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-95 variable light chain DNA

<400> 244

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacagta cccctcaggt cacttttggc 300

agcgggacac gactggagat caaa 324

<210> 245

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-1 variable light chain DNA

<400> 245

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta cccctatcgt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 246

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-6 variable light chain DNA

<400> 246

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatta 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag aagtacaaga cccctcaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 247

<211> 324

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-138 variable light chain DNA

<400> 247

gacatccaga tgacccagtc ccctcccgcc ctgtctgcat ctgtgggaga cagagtcacc 60

atcacttgcc gggccagtca agatattaat gactggttgg cctggtatca gcataaacct 120

gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttacta ctgtcaacag cgctacagta ccgaccaggt cacttttggc 300

caggggacac gactggagat caaa 324

<210> 248

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 variable heavy chain DNA

<400> 248

caggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggtggac 300

tacgctgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 249

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 variable heavy chain DNA

<400> 249

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggacgac 300

tacaacgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 250

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 variable heavy chain DNA

<400> 250

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggacgac 300

tacaacgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 251

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 variable heavy chain DNA

<400> 251

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aaggatcgac 300

tacgaggatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 252

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 variable heavy chain DNA

<400> 252

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggtgagc 300

tacgacgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 253

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 variable heavy chain DNA

<400> 253

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aaggatcgac 300

tacgaggatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 254

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 variable heavy chain DNA

<400> 254

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ttcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggtggac 300

taccaggagg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 255

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 variable heavy chain DNA

<400> 255

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggacgac 300

tacaacgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 256

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 variable heavy chain DNA

<400> 256

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggtggac 300

tacaacgagg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 257

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 variable heavy chain DNA

<400> 257

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggacgac 300

tacgaggatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 258

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 variable heavy chain DNA

<400> 258

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggtgagc 300

tacgctgagg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 259

<211> 354

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 variable heavy chain DNA

<400> 259

gaggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120

ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180

gcagactctg tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240

ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc aagggacgac 300

tacgacgatg catttgatat ctggggccag ggcaccctgg tcaccgtctc gagc 354

<210> 260

<211> 327

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7 variable light chain DNA

<400> 260

gctagcgaca tccagatgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 60

gtcaccatcg cttgccgggc aggtcagagc attggcacct atttaaattg gtatcagcag 120

aaaccaggga aagcccctaa actcctgatc tatgttgcat ccagtttgca aagtggggtc 180

ccgtcacggt tcagtggcag tggatctggg acagaattca ctctcaccat cagcagtctg 240

caacctgaag attttgcaac ttactactgt caacagagtt acagtaccct cctcactttc 300

ggcagaggga ccaaggtgga aatcaaa 327

<210> 261

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-10 variable light chain DNA

<400> 261

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag accgccagta ccctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 262

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-15 variable light chain DNA

<400> 262

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtgccgaca ccctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 263

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-17 variable light chain DNA

<400> 263

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtggcagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 264

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-18 variable light chain DNA

<400> 264

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggtaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtgccagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 265

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-19 variable light chain DNA

<400> 265

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtgccagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 266

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-20 variable light chain DNA

<400> 266

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agttacagtg gcctcgacac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 267

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-22 variable light chain DNA

<400> 267

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtgccagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 268

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-23 variable light chain DNA

<400> 268

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agttacagtg gcctcgacac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 269

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-42 variable light chain DNA

<400> 269

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtagcagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 270

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-3 variable light chain DNA

<400> 270

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agttacagtg gcctcgacac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 271

<211> 321

<212> DNA

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-7-61 variable light chain DNA

<400> 271

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcgcttgcc gggcaggtca gagcattggc acctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccgtca 180

cggttcagtg gcagtggatc tgggacagaa ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtgccagtg agctcctcac tttcggcaga 300

gggaccaagg tggaaatcaa a 321

<210> 272

<211> 209

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Human variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) -also known as TRDV1

<400> 272

Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser Met Pro Val Arg

1 5 10 15

Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser Trp Trp Ser Tyr

20 25 30

Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu Met Ile Phe Leu

35 40 45

Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser Gly Arg Tyr Ser

50 55 60

Val Asn Phe Lys Lys Ala Ala Lys Ser Val Ala Leu Thr Ile Ser Ala

65 70 75 80

Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala Leu Gly Glu Ser

85 90 95

Leu Thr Arg Ala Asp Lys Leu Ile Phe Gly Lys Gly Thr Arg Val Thr

100 105 110

Val Glu Pro Asn Ile Gln Asn Pro Asp Pro Ala Val Tyr Gln Leu Arg

115 120 125

Asp Ser Lys Ser Ser Asp Lys Ser Val Cys Leu Phe Thr Asp Phe Asp

130 135 140

Ser Gln Thr Asn Val Ser Gln Ser Lys Asp Ser Asp Val Tyr Ile Thr

145 150 155 160

Asp Lys Thr Val Leu Asp Met Arg Ser Met Asp Phe Lys Ser Asn Ser

165 170 175

Ala Val Ala Trp Ser Asn Lys Ser Asp Phe Ala Cys Ala Asn Ala Phe

180 185 190

Asn Asn Ser Ile Ile Pro Glu Asp Thr Phe Phe Pro Ser Pro Glu Ser

195 200 205

Ser

<210> 273

<211> 117

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 variable heavy chain

<400> 273

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Ser Pro Ile Glu Leu Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 274

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 variable heavy chain

<400> 274

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Asn Tyr Leu Asn Ala Phe Asp Ile Trp Gly Arg Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 275

<211> 117

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 variable heavy chain

<400> 275

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Gly Gly Gly Thr Thr Tyr Ser Ser Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ser Gly Val Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 276

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 variable heavy chain

<400> 276

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala

50 55 60

Val Ser Val Arg Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Ser Trp Asn Asp Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 277

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 variable heavy chain

<400> 277

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Val Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala

50 55 60

Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Asp Tyr Tyr Tyr Ser Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 278

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 variable heavy chain

<400> 278

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Trp Asn Asp Ala Phe Asp Val Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 279

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 variable heavy chain

<400> 279

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Gly Ser Lys Trp Tyr Asn Glu Tyr Ala

50 55 60

Leu Ser Val Lys Ser Arg Ile Ile Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Asp Tyr Tyr Tyr Ser Met Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 280

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 variable heavy chain

<400> 280

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Trp Ser Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 281

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 variable heavy chain

<400> 281

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Trp Asn Asp Ala Phe Asp Ile Trp Gly Arg Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 282

<211> 110

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 variable light chain

<400> 282

Ala Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro

1 5 10 15

Gly Lys Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Gln

20 25 30

Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Met Leu Val

35 40 45

Ile Tyr Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser

50 55 60

Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu

65 70 75 80

Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser

85 90 95

Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 283

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 variable light chain

<400> 283

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Ser Leu Ser

20 25 30

Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 284

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 variable light chain

<400> 284

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg

20 25 30

Thr Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Lys Arg Tyr

85 90 95

Pro Pro Thr Phe Gly Leu Gly Thr Lys Val Glu Ile Lys

100 105

<210> 285

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 variable light chain

<400> 285

Ala Ser Asp Ile Val Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser

1 5 10 15

Ile Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Gly Gln Ser Ile Ser

20 25 30

Thr Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 286

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 variable light chain

<400> 286

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

20 25 30

Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser His Ser His

85 90 95

Pro Pro Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 287

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 variable light chain

<400> 287

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

20 25 30

Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Asp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 288

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 variable light chain

<400> 288

Ala Ser Asp Ile Val Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser

1 5 10 15

Ile Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Gly Gln Ser Ile Ser

20 25 30

Thr Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Ala Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Val Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 289

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 variable light chain

<400> 289

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser

20 25 30

Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 290

<211> 109

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 variable light chain

<400> 290

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

20 25 30

Ser His Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Ala Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 291

<211> 13

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> VH-VL linker

<400> 291

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

1 5 10

<210> 292

<211> 12

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Optional scFv tag sequence (kappa sequence+His tag)

<400> 292

Arg Thr Ala Ala Ala Ser Ala His His His His His

1 5 10

<210> 293

<211> 37

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Optional scFv tag sequence (kappa sequence+His and FLAG tag)

<400> 293

Arg Thr Ala Ala Ala Ser Ala His His His His His His Lys Leu Asp

1 5 10 15

Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys

20 25 30

Asp Asp Asp Asp Lys

35

<210> 294

<211> 13

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Optional scFv tag sequence (lambda sequence+His tag)

<400> 294

Gly Gln Pro Ala Ala Ala Ser Ala His His His His His

1 5 10

<210> 295

<211> 38

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Optional scFv tag sequence (lambda sequence+His and FLAG tag)

<400> 295

Gly Gln Pro Ala Ala Ala Ser Ala His His His His His His Lys Leu

1 5 10 15

Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr

20 25 30

Lys Asp Asp Asp Asp Lys

35

<210> 296

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Constant domain (RTVAAPS sequence replaces the AD3 and AD4 full light constant chain of RTAAAPS)

<400> 296

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 297

<211> 330

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Constant domain (AD 3 full heavy chain constant domain)

<400> 297

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 298

<211> 330

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Constant domain (with AD4 full heavy chain constant domain (with L235A, G237A modification))

<400> 298

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 299

<211> 20

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> N-terminal leader sequence of V.delta.1 chain

<400> 299

Met Leu Phe Ser Ser Leu Leu Cys Val Phe Val Ala Phe Ser Tyr Ser

1 5 10 15

Gly Ser Ser Val

20

<210> 300

<211> 95

<212> PRT

<213> Chile (Homo Sapiens)

<220>

V region of <223> V.delta.1 chain

<400> 300

Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser Met Pro Val Arg

1 5 10 15

Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser Trp Trp Ser Tyr

20 25 30

Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu Met Ile Phe Leu

35 40 45

Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser Gly Arg Tyr Ser

50 55 60

Val Asn Phe Lys Lys Ala Ala Lys Ser Val Ala Leu Thr Ile Ser Ala

65 70 75 80

Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala Leu Gly Glu

85 90 95

<210> 301

<211> 16

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> J1 x 0-human delta single-stranded germline one of four regions encoded in

<400> 301

Thr Asp Lys Leu Ile Phe Gly Lys Gly Thr Arg Val Thr Val Glu Pro

1 5 10 15

<210> 302

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> J2 x 0-human delta single-stranded germline one of four regions encoded in

<400> 302

Leu Thr Ala Gln Leu Phe Phe Gly Lys Gly Thr Gln Leu Ile Val Glu

1 5 10 15

Pro

<210> 303

<211> 19

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> J3 x 0-human delta single-stranded germline one of four regions encoded in

<400> 303

Ser Trp Asp Thr Arg Gln Met Phe Phe Gly Thr Gly Ile Lys Leu Phe

1 5 10 15

Val Glu Pro

<210> 304

<211> 15

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> J4 x 0-human delta single-stranded germline one of four regions encoded in

<400> 304

Arg Pro Leu Ile Phe Gly Lys Gly Thr Tyr Leu Glu Val Gln Gln

1 5 10 15

<210> 305

<211> 153

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C1 x 0 containing C-terminal membrane proximal/transmembrane region

<220>

<221> misc_feature

<222> (1)..(1)

<223> Xaa can be any naturally occurring amino acid

<400> 305

Xaa Ser Gln Pro His Thr Lys Pro Ser Val Phe Val Met Lys Asn Gly

1 5 10 15

Thr Asn Val Ala Cys Leu Val Lys Glu Phe Tyr Pro Lys Asp Ile Arg

20 25 30

Ile Asn Leu Val Ser Ser Lys Lys Ile Thr Glu Phe Asp Pro Ala Ile

35 40 45

Val Ile Ser Pro Ser Gly Lys Tyr Asn Ala Val Lys Leu Gly Lys Tyr

50 55 60

Glu Ser Asn Ser Val Thr Cys Ser Val Gln His Asp Asn Lys Thr Val

65 70 75 80

His Ser Thr Asp Phe Glu Val Lys Thr Asp Ser Thr Asp His Val Lys

85 90 95

Pro Lys Glu Thr Glu Asn Thr Lys Gln Pro Ser Lys Ser Cys His Lys

100 105 110

Pro Lys Ala Ile Val His Thr Glu Lys Val Asn Met Met Ser Leu Thr

115 120 125

Val Leu Gly Leu Arg Met Leu Phe Ala Lys Thr Val Ala Val Asn Phe

130 135 140

Leu Leu Thr Ala Lys Leu Phe Phe Leu

145 150

<210> 306

<211> 254

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Human variable δ1 (vδ1) chain polymorphic variant of γδ T Cell Receptor (TCR) -also known as human TRDV1 polymorphic variant

<400> 306

Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser Met Pro Val Arg

1 5 10 15

Lys Ala Val Thr Leu Asn Cys Leu Tyr Glu Thr Ser Trp Trp Ser Tyr

20 25 30

Tyr Ile Phe Trp Tyr Lys Gln Leu Pro Ser Lys Glu Met Ile Phe Leu

35 40 45

Ile Arg Gln Gly Ser Asp Glu Gln Asn Ala Lys Ser Gly Arg Tyr Ser

50 55 60

Val Asn Phe Lys Lys Ala Val Lys Ser Val Ala Leu Thr Ile Ser Ala

65 70 75 80

Leu Gln Leu Glu Asp Ser Ala Lys Tyr Phe Cys Ala Leu Gly Glu Ser

85 90 95

Leu Thr Arg Ala Asp Lys Leu Ile Phe Gly Lys Gly Thr Arg Val Thr

100 105 110

Val Glu Pro Asn Ile Gln Asn Pro Asp Pro Ala Val Tyr Gln Leu Arg

115 120 125

Asp Ser Lys Ser Ser Asp Lys Ser Val Cys Leu Phe Thr Asp Phe Asp

130 135 140

Ser Gln Thr Asn Val Ser Gln Ser Lys Asp Ser Asp Val Tyr Ile Thr

145 150 155 160

Asp Lys Thr Val Leu Asp Met Arg Ser Met Asp Phe Lys Ser Asn Ser

165 170 175

Ala Val Ala Trp Ser Asn Lys Ser Asp Phe Ala Cys Ala Asn Ala Phe

180 185 190

Asn Asn Ser Ile Ile Pro Glu Asp Thr Phe Phe Pro Ser Pro Glu Ser

195 200 205

Ser Cys Thr Thr Ala Pro Ser Ala Gln Leu Lys Lys Lys Leu Gln Ala

210 215 220

Leu Lys Lys Lys Asn Ala Gln Leu Lys Trp Lys Leu Gln Ala Leu Lys

225 230 235 240

Lys Lys Leu Ala Gln Gly Ser Gly His His His His His His

245 250

<210> 307

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Constant domain (AD 3 and AD4 full light constant chain with RTAAAPS sequences)

<400> 307

Arg Thr Ala Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 308

<211> 116

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Cynomolgus monkey variable delta 1 (V delta 1) chain of gamma delta T Cell Receptor (TCR) -also known as cynomolgus monkey TRDV1 (minus leader sequence)

<400> 308

Ala Gln Lys Val Thr Gln Ala Gln Ser Ser Val Ser Met Pro Val Glu

1 5 10 15

Lys Ala Val Thr Leu Asn Cys Gln Tyr Glu Thr Ser Ser Trp Ser Tyr

20 25 30

Asp Leu Phe Trp Tyr Lys Gln Leu Pro Gly Lys Glu Met Ile Phe Leu

35 40 45

Ile His Gln Gly Ser Ser Gln Gln Asn Ala Arg Asn Gly Arg Tyr Ser

50 55 60

Val Asn Phe Gln Lys Ala Ala Ser Ser Ile Thr Leu Thr Ile Ser Ala

65 70 75 80

Leu Gln Leu Glu Asp Ser Ala Thr Tyr Phe Cys Ala

85 90

<210> 309

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Human variable gamma 4 chain (V gamma 4) of gamma delta T Cell Receptor (TCR) -also known as human TRGV4 (minus leader sequence)

<400> 309

Lys Ser Ser Asn Leu Glu Gly Arg Thr Lys Ser Val Ile Arg Gln Thr

1 5 10 15

Gly Ser Ser Ala Glu Ile Thr Cys Asp Leu Ala Glu Gly Ser Thr Gly

20 25 30

Tyr Ile His Trp Tyr Leu His Gln Glu Gly Lys Ala Pro Gln Arg Leu

35 40 45

Leu Tyr Tyr Asp Ser Tyr Thr Ser Ser Val Val Leu Glu Ser Gly Ile

50 55 60

Ser Pro Gly Lys Tyr Asp Thr Tyr Gly Ser Thr Arg Lys Asn Leu Arg

65 70 75 80

Met Ile Leu Arg Asn Leu Ile Glu Asn Asp Ser Gly Val Tyr Tyr Cys

85 90 95

Ala Thr Trp Asp Gly

100

<210> 310

<211> 115

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Human variable delta 2 (V delta 2) chain of gamma delta T Cell Receptor (TCR) -also known as TRDV2 (minus leader sequence)

<400> 310

Ala Ile Glu Leu Val Pro Glu His Gln Thr Val Pro Val Ser Ile Gly

1 5 10 15

Val Pro Ala Thr Leu Arg Cys Ser Met Lys Gly Glu Ala Ile Gly Asn

20 25 30

Tyr Tyr Ile Asn Trp Tyr Arg Lys Thr Gln Gly Asn Thr Met Thr Phe

35 40 45

Ile Tyr Arg Glu Lys Asp Ile Tyr Gly Pro Gly Phe Lys Asp Asn Phe

50 55 60

Gln Gly Asp Ile Asp Ile Ala Lys Asn Leu Ala Val Leu Lys Ile Leu

65 70 75 80

Ala Pro Ser Glu Arg Asp Glu Gly Ser Tyr Tyr Cys Ala Cys Asp Thr

85 90 95

<210> 311

<211> 113

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Human variable delta 3 (V delta 3) chain of gamma delta T Cell Receptor (TCR) -also known as TRDV3 (minus leader sequence)

<400> 311

Asp Lys Val Thr Gln Ser Ser Pro Asp Gln Thr Val Ala Ser Gly Ser

1 5 10 15

Glu Val Val Leu Leu Cys Thr Tyr Asp Thr Val Tyr Ser Asn Pro Asp

20 25 30

Leu Phe Trp Tyr Arg Ile Arg Pro Asp Tyr Ser Phe Gln Phe Val Phe

35 40 45

Tyr Gly Asp Asn Ser Arg Ser Glu Gly Ala Asp Phe Thr Gln Gly Arg

50 55 60

Phe Ser Val Lys His Ile Leu Thr Gln Lys Ala Phe His Leu Val Ile

65 70 75 80

Ser Pro Val Arg Thr Glu Asp Ser Ala Thr Tyr Tyr Cys Ala Phe

85 90 95

<210> 312

<211> 119

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E01 variable heavy chain

<400> 312

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr

20 25 30

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe

50 55 60

Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg His Gln Val Asp Thr Arg Thr Ala Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 313

<211> 112

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E01 variable light chain

<400> 313

Ala Ser Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro

1 5 10 15

Gly Gln Ser Val Thr Ile Ser Cys Thr Gly Thr Arg Ser Asp Val Gly

20 25 30

Gly Tyr Asn Tyr Val Ser Trp Tyr Gln His His Pro Gly Lys Ala Pro

35 40 45

Lys Leu Met Ile Tyr Glu Val Ser Asn Arg Pro Ser Gly Val Ser Asn

50 55 60

Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser

65 70 75 80

Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr

85 90 95

Ser Thr Ser Thr Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 314

<211> 250

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ScFv anti-CD 19 binding Module

<400> 314

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Gln Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Leu Thr

130 135 140

Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile

145 150 155 160

Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu

165 170 175

Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

180 185 190

Asp Ala Ser Asn Leu Val Ser Gly Ile Pro Pro Arg Phe Ser Gly Ser

195 200 205

Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Lys Val

210 215 220

Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr Glu Asp Pro Trp Thr

225 230 235 240

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

245 250

<210> 315

<211> 931

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G04 x anti-CD 19 multispecific (full Length)

<400> 315

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

20 25 30

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr

100 105 110

Ala Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln Gln Ser

210 215 220

Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala

225 230 235 240

Ile Ser Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile

245 250 255

Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr

260 265 270

Arg Ser Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile

275 280 285

Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn

290 295 300

Ser Val Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp

305 310 315 320

Ser Gly Tyr Val Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser

325 330 335

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

340 345 350

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

355 360 365

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

370 375 380

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

385 390 395 400

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

405 410 415

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

420 425 430

Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

435 440 445

Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro

450 455 460

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

465 470 475 480

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

485 490 495

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

500 505 510

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

515 520 525

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

530 535 540

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

545 550 555 560

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

565 570 575

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

580 585 590

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

595 600 605

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

610 615 620

Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

625 630 635 640

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

645 650 655

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly

660 665 670

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser

675 680 685

Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val Lys Ile Ser Cys Lys

690 695 700

Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gln

705 710 715 720

Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Gln Ile Trp Pro Gly Asp

725 730 735

Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr

740 745 750

Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Ala

755 760 765

Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr

770 775 780

Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr

785 790 795 800

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

805 810 815

Gly Gly Gly Ser Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala

820 825 830

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser

835 840 845

Val Asp Tyr Asp Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro

850 855 860

Gly Gln Pro Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Val Ser

865 870 875 880

Gly Ile Pro Pro Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

885 890 895

Leu Asn Ile His Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys

900 905 910

Gln Gln Ser Thr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

915 920 925

Glu Ile Lys

930

<210> 316

<211> 928

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E07 x anti-CD 19 multispecific (full Length)

<400> 316

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly

20 25 30

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val Glu Ser Gly

210 215 220

Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

225 230 235 240

Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala

245 250 255

Pro Gly Lys Gly Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser

260 265 270

Thr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

275 280 285

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

290 295 300

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Asp Tyr Ala Asp Ala

305 310 315 320

Phe Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

325 330 335

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr

340 345 350

Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

355 360 365

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

370 375 380

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

385 390 395 400

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile

405 410 415

Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val

420 425 430

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

435 440 445

Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

450 455 460

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

465 470 475 480

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

485 490 495

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

500 505 510

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

515 520 525

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

530 535 540

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

545 550 555 560

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr

565 570 575

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

580 585 590

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

595 600 605

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

610 615 620

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

625 630 635 640

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

645 650 655

Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly

660 665 670

Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu

675 680 685

Leu Val Arg Pro Gly Ser Ser Val Lys Ile Ser Cys Lys Ala Ser Gly

690 695 700

Tyr Ala Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gln Arg Pro Gly

705 710 715 720

Gln Gly Leu Glu Trp Ile Gly Gln Ile Trp Pro Gly Asp Gly Asp Thr

725 730 735

Asn Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu

740 745 750

Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Ala Ser Glu Asp

755 760 765

Ser Ala Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg

770 775 780

Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

785 790 795 800

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

805 810 815

Ser Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu

820 825 830

Gly Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr

835 840 845

Asp Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro

850 855 860

Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Val Ser Gly Ile Pro

865 870 875 880

Pro Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile

885 890 895

His Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gln Gln Ser

900 905 910

Thr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

915 920 925

<210> 317

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 HCDR1

<400> 317

Gly Phe Thr Val Ser Ser Asn Tyr

1 5

<210> 318

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 HCDR2

<400> 318

Ile Tyr Ser Gly Gly Ser Thr

1 5

<210> 319

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 HCDR3

<400> 319

Pro Ile Glu Leu Gly Ala Phe Asp Ile

1 5

<210> 320

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 HCDR1

<400> 320

Gly Phe Thr Phe Ser Asp Tyr Tyr

1 5

<210> 321

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 HCDR2

<400> 321

Ile Ser Ser Ser Gly Ser Thr Ile

1 5

<210> 322

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 HCDR3

<400> 322

Glu Asn Tyr Leu Asn Ala Phe Asp Ile

1 5

<210> 323

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 HCDR1

<400> 323

Gly Phe Thr Phe Ser Ser Tyr Ala

1 5

<210> 324

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 HCDR2

<400> 324

Ile Ser Gly Gly Gly Gly Thr Thr

1 5

<210> 325

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 HCDR3

<400> 325

Asp Ser Gly Val Ala Phe Asp Ile

1 5

<210> 326

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 HCDR1

<400> 326

Gly Asp Ser Val Ser Ser Asn Ser Ala Ala

1 5 10

<210> 327

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 HCDR2

<400> 327

Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn

1 5

<210> 328

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 HCDR3

<400> 328

Ser Trp Asn Asp Ala Phe Asp Ile

1 5

<210> 329

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 HCDR1

<400> 329

Gly Asp Ser Val Ser Ser Asn Ser Ala Ala

1 5 10

<210> 330

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 HCDR2

<400> 330

Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn

1 5

<210> 331

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 HCDR3

<400> 331

Asp Tyr Tyr Tyr Ser Met Asp Val

1 5

<210> 332

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 HCDR1

<400> 332

Gly Phe Thr Phe Ser Asp Tyr Tyr

1 5

<210> 333

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 HCDR2

<400> 333

Ile Ser Ser Ser Gly Ser Thr Ile

1 5

<210> 334

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 HCDR3

<400> 334

His Ser Trp Asn Asp Ala Phe Asp Val

1 5

<210> 335

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 HCDR1

<400> 335

Gly Asp Ser Val Ser Ser Asn Ser Ala Ala

1 5 10

<210> 336

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 HCDR2

<400> 336

Thr Tyr Tyr Gly Ser Lys Trp Tyr Asn

1 5

<210> 337

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 HCDR3

<400> 337

Asp Tyr Tyr Tyr Ser Met Asp Val

1 5

<210> 338

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 HCDR1

<400> 338

Gly Phe Thr Phe Ser Asp Tyr Tyr

1 5

<210> 339

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 HCDR2

<400> 339

Ile Ser Ser Ser Gly Ser Thr Ile

1 5

<210> 340

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 HCDR3

<400> 340

His Ser Trp Ser Asp Ala Phe Asp Ile

1 5

<210> 341

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 HCDR1

<400> 341

Gly Phe Thr Phe Ser Asp Tyr Tyr

1 5

<210> 342

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 HCDR2

<400> 342

Ile Ser Ser Ser Gly Ser Thr Ile

1 5

<210> 343

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 HCDR3

<400> 343

His Ser Trp Asn Asp Ala Phe Asp Ile

1 5

<210> 344

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E01 HCDR1

<400> 344

Gly Tyr Ser Phe Thr Ser Tyr Trp

1 5

<210> 345

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E02 HCDR2

<400> 345

Ile Tyr Pro Gly Asp Ser Asp Thr

1 5

<210> 346

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E02 HCDR3

<400> 346

His Gln Val Asp Thr Arg Thr Ala Asp Tyr

1 5 10

<210> 347

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 LCDR1

<400> 347

Asn Ile Gly Ser Gln Ser

1 5

<210> 348

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 LCDR2

<400> 348

Tyr Asp Ser

1

<210> 349

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 LCDR3

<400> 349

Gln Val Trp Asp Ser Ser Ser Asp His Val Val

1 5 10

<210> 350

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 LCDR1

<400> 350

Gln Ser Leu Ser Asn Tyr

1 5

<210> 351

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 LCDR2

<400> 351

Ala Ala Ser

1

<210> 352

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B07 LCDR3

<400> 352

Gln Gln Ser Tyr Ser Thr Pro Leu Thr

1 5

<210> 353

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 LCDR1

<400> 353

Gln Asn Ile Arg Thr Trp

1 5

<210> 354

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 LCDR2

<400> 354

Asp Ala Ser

1

<210> 355

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C05 LCDR3

<400> 355

Gln Gln Phe Lys Arg Tyr Pro Pro Thr

1 5

<210> 356

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 LCDR1

<400> 356

Gln Ser Ile Ser Thr Trp

1 5

<210> 357

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 LCDR2

<400> 357

Asp Ala Ser

1

<210> 358

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E04 LCDR3

<400> 358

Gln Gln Ser Tyr Ser Thr Pro Leu Thr

1 5

<210> 359

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 LCDR1

<400> 359

Gln Ser Ile Ser Ser Trp

1 5

<210> 360

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 LCDR2

<400> 360

Asp Ala Ser

1

<210> 361

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> F07 LCDR3

<400> 361

Gln Gln Ser His Ser His Pro Pro Thr

1 5

<210> 362

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 LCDR1

<400> 362

Gln Ser Ile Ser Ser Tyr

1 5

<210> 363

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 LCDR2

<400> 363

Ala Ala Ser

1

<210> 364

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G06 LCDR3

<400> 364

Gln Gln Ser Tyr Ser Thr Pro Asp Thr

1 5

<210> 365

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 LCDR1

<400> 365

Gln Ser Ile Ser Thr Trp

1 5

<210> 366

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 LCDR2

<400> 366

Asp Ala Ser

1

<210> 367

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G09 LCDR3

<400> 367

Gln Gln Ser Tyr Ser Thr Pro Val Thr

1 5

<210> 368

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 LCDR1

<400> 368

Gln Asp Ile Ser Asn Tyr

1 5

<210> 369

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 LCDR2

<400> 369

Asp Ala Ser

1

<210> 370

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> B09 LCDR3

<400> 370

Gln Gln Ser Tyr Ser Thr Pro Leu Thr

1 5

<210> 371

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 LCDR1

<400> 371

Gln Ser Ile Ser Ser His

1 5

<210> 372

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 LCDR2

<400> 372

Ala Ala Ser

1

<210> 373

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G10 LCDR3

<400> 373

Gln Gln Ser Tyr Ser Thr Leu Leu Thr

1 5

<210> 374

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E01 LCDR1

<400> 374

Arg Ser Asp Val Gly Gly Tyr Asn Tyr

1 5

<210> 375

<211> 3

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E02 LCDR2

<400> 375

Glu Val Ser

1

<210> 376

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E02 LCDR3

<400> 376

Ser Ser Tyr Thr Ser Thr Ser Thr Leu Val

1 5 10

<210> 377

<211> 661

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Anti-chicken lysozyme x FS1-67 Fcab (EGFR binding domain)

<400> 377

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His

20 25 30

Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Tyr Thr Thr Thr Leu Ala Asp Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln His Phe Trp Ser Thr

85 90 95

Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln Glu Ser Gly

210 215 220

Pro Gly Leu Val Arg Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val

225 230 235 240

Ser Gly Ser Thr Phe Ser Gly Tyr Gly Val Asn Trp Val Arg Gln Pro

245 250 255

Pro Gly Arg Gly Leu Glu Trp Ile Gly Met Ile Trp Gly Asp Gly Asn

260 265 270

Thr Asp Tyr Asn Ser Ala Leu Lys Ser Arg Val Thr Met Leu Val Asp

275 280 285

Thr Ser Lys Asn Gln Phe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala

290 295 300

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Asp Tyr Arg Leu Asp

305 310 315 320

Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser Ser Ala Ser Thr Lys

325 330 335

Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly

340 345 350

Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro

355 360 365

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

370 375 380

Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val

385 390 395 400

Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn

405 410 415

Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro

420 425 430

Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

435 440 445

Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

450 455 460

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

465 470 475 480

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

485 490 495

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

500 505 510

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

515 520 525

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

530 535 540

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

545 550 555 560

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp Asp Gly

565 570 575

Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

580 585 590

Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr

595 600 605

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

610 615 620

Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys

625 630 635 640

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

645 650 655

Ser Leu Ser Pro Gly

660

<210> 378

<211> 662

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parent C08 x FS1-67 (LAGA)

<400> 378

Ala Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro

1 5 10 15

Gly Lys Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Gln

20 25 30

Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Met Leu Val

35 40 45

Ile Tyr Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser

50 55 60

Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu

65 70 75 80

Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser

85 90 95

Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

100 105 110

Pro Ala Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

115 120 125

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

130 135 140

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys

145 150 155 160

Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr

165 170 175

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

180 185 190

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

195 200 205

Thr Val Ala Pro Thr Glu Cys Ser Glu Val Gln Leu Leu Glu Ser Gly

210 215 220

Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala

225 230 235 240

Ser Gly Phe Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala

245 250 255

Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser

260 265 270

Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

275 280 285

Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

290 295 300

Asp Thr Ala Val Tyr Tyr Cys Ala Ser Pro Ile Glu Leu Gly Ala Phe

305 310 315 320

Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

325 330 335

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser

340 345 350

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu

355 360 365

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His

370 375 380

Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser

385 390 395 400

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys

405 410 415

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu

420 425 430

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

435 440 445

Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

450 455 460

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

465 470 475 480

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

485 490 495

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

500 505 510

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

515 520 525

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

530 535 540

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

545 550 555 560

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp Asp

565 570 575

Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

580 585 590

Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys

595 600 605

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

610 615 620

Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser

625 630 635 640

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

645 650 655

Leu Ser Leu Ser Pro Gly

660

<210> 379

<211> 666

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parent G04 x FS1-67 (LAGA)

<400> 379

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

20 25 30

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr

100 105 110

Ala Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln Gln Ser

210 215 220

Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala

225 230 235 240

Ile Ser Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile

245 250 255

Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr

260 265 270

Arg Ser Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile

275 280 285

Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn

290 295 300

Ser Val Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp

305 310 315 320

Ser Gly Tyr Val Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser

325 330 335

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

340 345 350

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

355 360 365

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

370 375 380

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

385 390 395 400

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

405 410 415

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

420 425 430

Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

435 440 445

Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro

450 455 460

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

465 470 475 480

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

485 490 495

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

500 505 510

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

515 520 525

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

530 535 540

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

545 550 555 560

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

565 570 575

Glu Thr Asp Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe

580 585 590

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu

595 600 605

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

610 615 620

Phe Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly

625 630 635 640

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

645 650 655

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

660 665

<210> 380

<211> 663

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parent E07 x FS1-67 (LAGA)

<400> 380

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly

20 25 30

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val Glu Ser Gly

210 215 220

Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

225 230 235 240

Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala

245 250 255

Pro Gly Lys Gly Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser

260 265 270

Thr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

275 280 285

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

290 295 300

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Asp Tyr Ala Asp Ala

305 310 315 320

Phe Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

325 330 335

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr

340 345 350

Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

355 360 365

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

370 375 380

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

385 390 395 400

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile

405 410 415

Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val

420 425 430

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

435 440 445

Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

450 455 460

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

465 470 475 480

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

485 490 495

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

500 505 510

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

515 520 525

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

530 535 540

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

545 550 555 560

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp

565 570 575

Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

580 585 590

Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr

595 600 605

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

610 615 620

Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe

625 630 635 640

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

645 650 655

Ser Leu Ser Leu Ser Pro Gly

660

<210> 381

<211> 917

<212> PRT

<213> Artificial sequence (ARTIFICIAL SEQUENCE)

<220>

<223> Anti-chicken lysozyme x linker cetuximab scFV (EGFR binding domain as Morisen at the C-terminus of CH3

<400> 381

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His

20 25 30

Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Tyr Thr Thr Thr Leu Ala Asp Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln His Phe Trp Ser Thr

85 90 95

Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln Glu Ser Gly

210 215 220

Pro Gly Leu Val Arg Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val

225 230 235 240

Ser Gly Ser Thr Phe Ser Gly Tyr Gly Val Asn Trp Val Arg Gln Pro

245 250 255

Pro Gly Arg Gly Leu Glu Trp Ile Gly Met Ile Trp Gly Asp Gly Asn

260 265 270

Thr Asp Tyr Asn Ser Ala Leu Lys Ser Arg Val Thr Met Leu Val Asp

275 280 285

Thr Ser Lys Asn Gln Phe Ser Leu Arg Leu Ser Ser Val Thr Ala Ala

290 295 300

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Asp Tyr Arg Leu Asp

305 310 315 320

Tyr Trp Gly Gln Gly Ser Leu Val Thr Val Ser Ser Ala Ser Thr Lys

325 330 335

Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly

340 345 350

Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro

355 360 365

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

370 375 380

Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val

385 390 395 400

Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn

405 410 415

Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro

420 425 430

Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

435 440 445

Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

450 455 460

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

465 470 475 480

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

485 490 495

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

500 505 510

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

515 520 525

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

530 535 540

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

545 550 555 560

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn

565 570 575

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

580 585 590

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

595 600 605

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

610 615 620

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

625 630 635 640

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

645 650 655

Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

660 665 670

Gly Gly Gly Ser Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val

675 680 685

Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser

690 695 700

Leu Thr Asn Tyr Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly

705 710 715 720

Leu Glu Trp Leu Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn

725 730 735

Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser

740 745 750

Gln Val Phe Phe Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile

755 760 765

Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr

770 775 780

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser

785 790 795 800

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Leu Leu Thr Gln

805 810 815

Ser Pro Val Ile Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe Ser

820 825 830

Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn Ile His Trp Tyr Gln Gln

835 840 845

Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Glu Ser

850 855 860

Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

865 870 875 880

Phe Thr Leu Ser Ile Asn Ser Val Glu Ser Glu Asp Ile Ala Asp Tyr

885 890 895

Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr Thr Phe Gly Ala Gly Thr

900 905 910

Lys Leu Glu Leu Lys

915

<210> 382

<211> 918

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parental C08 x linker cetuximab scFV (EGFR binding domain as Morisen at C-terminus of CH 3)

<400> 382

Ala Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro

1 5 10 15

Gly Lys Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Gln

20 25 30

Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Met Leu Val

35 40 45

Ile Tyr Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser

50 55 60

Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu

65 70 75 80

Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser

85 90 95

Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

100 105 110

Pro Ala Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

115 120 125

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

130 135 140

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys

145 150 155 160

Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr

165 170 175

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

180 185 190

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

195 200 205

Thr Val Ala Pro Thr Glu Cys Ser Glu Val Gln Leu Leu Glu Ser Gly

210 215 220

Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala

225 230 235 240

Ser Gly Phe Thr Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala

245 250 255

Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser

260 265 270

Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

275 280 285

Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu

290 295 300

Asp Thr Ala Val Tyr Tyr Cys Ala Ser Pro Ile Glu Leu Gly Ala Phe

305 310 315 320

Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

325 330 335

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser

340 345 350

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu

355 360 365

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His

370 375 380

Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser

385 390 395 400

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys

405 410 415

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu

420 425 430

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

435 440 445

Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

450 455 460

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

465 470 475 480

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

485 490 495

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

500 505 510

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

515 520 525

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

530 535 540

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

545 550 555 560

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys

565 570 575

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

580 585 590

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

595 600 605

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

610 615 620

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

625 630 635 640

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

645 650 655

Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

660 665 670

Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu

675 680 685

Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe

690 695 700

Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg Gln Ser Pro Gly Lys

705 710 715 720

Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr

725 730 735

Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys

740 745 750

Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala

755 760 765

Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala

770 775 780

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly

785 790 795 800

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Leu Leu Thr

805 810 815

Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe

820 825 830

Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn Ile His Trp Tyr Gln

835 840 845

Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Glu

850 855 860

Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr

865 870 875 880

Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser Glu Asp Ile Ala Asp

885 890 895

Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr Thr Phe Gly Ala Gly

900 905 910

Thr Lys Leu Glu Leu Lys

915

<210> 383

<211> 922

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parental G04 x linker cetuximab scFV (EGFR binding domain as Morisen at the C-terminus of CH 3)

<400> 383

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

20 25 30

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr

100 105 110

Ala Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln Gln Ser

210 215 220

Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala

225 230 235 240

Ile Ser Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile

245 250 255

Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr

260 265 270

Arg Ser Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile

275 280 285

Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn

290 295 300

Ser Val Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp

305 310 315 320

Ser Gly Tyr Val Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser

325 330 335

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

340 345 350

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

355 360 365

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

370 375 380

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

385 390 395 400

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

405 410 415

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

420 425 430

Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

435 440 445

Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro

450 455 460

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

465 470 475 480

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

485 490 495

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

500 505 510

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

515 520 525

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

530 535 540

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

545 550 555 560

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

565 570 575

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

580 585 590

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

595 600 605

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

610 615 620

Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

625 630 635 640

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

645 650 655

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly

660 665 670

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln Ser

675 680 685

Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys Thr

690 695 700

Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg Gln

705 710 715 720

Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly Gly

725 730 735

Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn Lys

740 745 750

Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln Ser

755 760 765

Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr Asp

770 775 780

Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala

785 790 795 800

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp

805 810 815

Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly Glu

820 825 830

Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn Ile

835 840 845

His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile Lys

850 855 860

Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser

865 870 875 880

Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser Glu

885 890 895

Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr Thr

900 905 910

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

915 920

<210> 384

<211> 919

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Parental E07 x linker cetuximab scFV (EGFR binding domain as Morisen at C-terminal of CH 3)

<400> 384

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly

20 25 30

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val Glu Ser Gly

210 215 220

Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

225 230 235 240

Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala

245 250 255

Pro Gly Lys Gly Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser

260 265 270

Thr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

275 280 285

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

290 295 300

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Asp Tyr Ala Asp Ala

305 310 315 320

Phe Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

325 330 335

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr

340 345 350

Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

355 360 365

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

370 375 380

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

385 390 395 400

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile

405 410 415

Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val

420 425 430

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

435 440 445

Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

450 455 460

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

465 470 475 480

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

485 490 495

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

500 505 510

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

515 520 525

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

530 535 540

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

545 550 555 560

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr

565 570 575

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

580 585 590

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

595 600 605

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

610 615 620

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

625 630 635 640

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

645 650 655

Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly

660 665 670

Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln Ser Gly Pro Gly

675 680 685

Leu Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys Thr Val Ser Gly

690 695 700

Phe Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg Gln Ser Pro Gly

705 710 715 720

Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly Gly Asn Thr Asp

725 730 735

Tyr Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser

740 745 750

Lys Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln Ser Asn Asp Thr

755 760 765

Ala Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe

770 775 780

Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly

785 790 795 800

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Leu Leu

805 810 815

Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly Glu Arg Val Ser

820 825 830

Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn Ile His Trp Tyr

835 840 845

Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser

850 855 860

Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly

865 870 875 880

Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser Glu Asp Ile Ala

885 890 895

Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr Thr Phe Gly Ala

900 905 910

Gly Thr Lys Leu Glu Leu Lys

915

<210> 385

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> IgG1 CH1-CH2-CH3 FS-167 Fcab

<400> 385

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Thr Asp Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 386

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> IgG1 CH1-CH2-CH3 FS-167 Fcab (LAGA)

<400> 386

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Thr Asp Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 387

<211> 241

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ScFV cetuximab (in Morisebody format)

<400> 387

Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln

1 5 10 15

Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr

20 25 30

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr

50 55 60

Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe

65 70 75 80

Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Leu Leu Thr Gln Ser Pro Val Ile

130 135 140

Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg Ala Ser

145 150 155 160

Gln Ser Ile Gly Thr Asn Ile His Trp Tyr Gln Gln Arg Thr Asn Gly

165 170 175

Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile

180 185 190

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser

195 200 205

Ile Asn Ser Val Glu Ser Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln

210 215 220

Asn Asn Asn Trp Pro Thr Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu

225 230 235 240

Lys

<210> 388

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x FS1-67 EGFR (LAGA)

<400> 388

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Thr

565 570 575

Asp Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 389

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x LEE (LAGA)

<400> 389

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Glu Glu Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 390

<211> 257

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Folate receptor alpha

<400> 390

Met Ala Gln Arg Met Thr Thr Gln Leu Leu Leu Leu Leu Val Trp Val

1 5 10 15

Ala Val Val Gly Glu Ala Gln Thr Arg Ile Ala Trp Ala Arg Thr Glu

20 25 30

Leu Leu Asn Val Cys Met Asn Ala Lys His His Lys Glu Lys Pro Gly

35 40 45

Pro Glu Asp Lys Leu His Glu Gln Cys Arg Pro Trp Arg Lys Asn Ala

50 55 60

Cys Cys Ser Thr Asn Thr Ser Gln Glu Ala His Lys Asp Val Ser Tyr

65 70 75 80

Leu Tyr Arg Phe Asn Trp Asn His Cys Gly Glu Met Ala Pro Ala Cys

85 90 95

Lys Arg His Phe Ile Gln Asp Thr Cys Leu Tyr Glu Cys Ser Pro Asn

100 105 110

Leu Gly Pro Trp Ile Gln Gln Val Asp Gln Ser Trp Arg Lys Glu Arg

115 120 125

Val Leu Asn Val Pro Leu Cys Lys Glu Asp Cys Glu Gln Trp Trp Glu

130 135 140

Asp Cys Arg Thr Ser Tyr Thr Cys Lys Ser Asn Trp His Lys Gly Trp

145 150 155 160

Asn Trp Thr Ser Gly Phe Asn Lys Cys Ala Val Gly Ala Ala Cys Gln

165 170 175

Pro Phe His Phe Tyr Phe Pro Thr Pro Thr Val Leu Cys Asn Glu Ile

180 185 190

Trp Thr His Ser Tyr Lys Val Ser Asn Tyr Ser Arg Gly Ser Gly Arg

195 200 205

Cys Ile Gln Met Trp Phe Asp Pro Ala Gln Gly Asn Pro Asn Glu Glu

210 215 220

Val Ala Arg Phe Tyr Ala Ala Ala Met Ser Gly Ala Gly Pro Trp Ala

225 230 235 240

Ala Trp Pro Phe Leu Leu Ser Leu Ala Leu Met Leu Leu Trp Leu Leu

245 250 255

Ser

<210> 391

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR (LEE) binding Module (LAGA)

<400> 391

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Glu Glu Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 392

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR (LEE) binding Module (IgG 1 wt)

<400> 392

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Glu Glu Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 393

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HER2 (LAGA)

<400> 393

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Phe

565 570 575

Phe Thr Tyr Trp Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Arg Arg Arg Trp Thr Ala Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 394

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> HER2 binding Module (LAGA)

<400> 394

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Phe Phe Thr Tyr Trp Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Arg Arg Arg Trp Thr Ala Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 395

<211> 329

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> HER2 binding Module (Ig WT)

<400> 395

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Phe Phe Thr Tyr Trp Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Arg Arg Arg Trp Thr Ala Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 396

<211> 498

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x CD19

<400> 396

Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp

20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Val Ser Gly Ile Pro Pro

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His

65 70 75 80

Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val

115 120 125

Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val

130 135 140

Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met

145 150 155 160

Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Gln

165 170 175

Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly

180 185 190

Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gln

195 200 205

Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg

210 215 220

Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp

225 230 235 240

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gln

245 250 255

Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr

260 265 270

Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys Ser

275 280 285

Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp

290 295 300

Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala Ala

305 310 315 320

Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln

325 330 335

Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val Tyr

340 345 350

Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln Gly

355 360 365

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

370 375 380

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala

385 390 395 400

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

405 410 415

Gln Asp Ile Asn Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys

420 425 430

Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val

435 440 445

Pro Leu Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr

450 455 460

Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

465 470 475 480

Lys Tyr Ser Ala Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu

485 490 495

Ile Lys

<210> 397

<211> 250

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 binding Module

<400> 397

Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp

20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Val Ser Gly Ile Pro Pro

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His

65 70 75 80

Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val

115 120 125

Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val

130 135 140

Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met

145 150 155 160

Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Gln

165 170 175

Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly

180 185 190

Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gln

195 200 205

Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg

210 215 220

Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp

225 230 235 240

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

245 250

<210> 398

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HER2 (wt IgG1)

<400> 398

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Phe

565 570 575

Phe Thr Tyr Trp Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Arg Arg Arg Trp Thr Ala Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 399

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x FS1-67 EGFR (wt)

<400> 399

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Thr

565 570 575

Asp Asp Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 400

<211> 664

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x LEE (wt)

<400> 400

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Glu Glu Gly Pro Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Thr Tyr Gly Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg Trp Tyr Lys Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 401

<211> 931

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x FAPα

<400> 401

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly

660 665 670

Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser Gly Ala

675 680 685

Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Thr Ser

690 695 700

Arg Tyr Thr Phe Thr Glu Tyr Thr Ile His Trp Val Arg Gln Ala Pro

705 710 715 720

Gly Gln Arg Leu Glu Trp Ile Gly Gly Ile Asn Pro Asn Asn Gly Ile

725 730 735

Pro Asn Tyr Asn Gln Lys Phe Lys Gly Arg Val Thr Ile Thr Val Asp

740 745 750

Thr Ser Ala Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu

755 760 765

Asp Thr Ala Val Tyr Tyr Cys Ala Arg Arg Arg Ile Ala Tyr Gly Tyr

770 775 780

Asp Glu Gly His Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr

785 790 795 800

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

805 810 815

Gly Ser Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser

820 825 830

Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu

835 840 845

Tyr Ser Arg Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro

850 855 860

Gly Gln Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser

865 870 875 880

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Phe Gly Thr Asp Phe Thr

885 890 895

Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys

900 905 910

Gln Gln Tyr Phe Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val

915 920 925

Glu Ile Lys

930

<210> 402

<211> 252

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-FAP alpha scFv binding Module

<400> 402

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Thr Ser Arg Tyr Thr Phe Thr Glu Tyr

20 25 30

Thr Ile His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile

35 40 45

Gly Gly Ile Asn Pro Asn Asn Gly Ile Pro Asn Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Val Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Arg Ile Ala Tyr Gly Tyr Asp Glu Gly His Ala Met Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr

130 135 140

Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile

145 150 155 160

Asn Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser Arg Asn Gln Lys Asn

165 170 175

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

180 185 190

Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser

195 200 205

Gly Ser Gly Phe Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

210 215 220

Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Phe Ser Tyr Pro

225 230 235 240

Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

245 250

<210> 403

<211> 919

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 x MSLN

<400> 403

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly

660 665 670

Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu Ser Gly Pro

675 680 685

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser

690 695 700

Gly Gly Ser Ile Asn Asn Asn Asn Tyr Tyr Trp Thr Trp Ile Arg Gln

705 710 715 720

His Pro Gly Lys Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly

725 730 735

Ser Thr Phe Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val

740 745 750

Asp Thr Ser Lys Thr Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala

755 760 765

Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Asp Thr Met Thr Gly

770 775 780

Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly

785 790 795 800

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln

805 810 815

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

820 825 830

Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Asn Tyr Leu Asn Trp

835 840 845

Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile Tyr Ala Ala

850 855 860

Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Arg Ser

865 870 875 880

Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe

885 890 895

Ala Ala Tyr Phe Cys Gln Gln Thr Tyr Ser Asn Pro Thr Phe Gly Gln

900 905 910

Gly Thr Lys Val Glu Val Lys

915

<210> 404

<211> 240

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-MSLN scFv binding module

<400> 404

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Asn

20 25 30

Asn Tyr Tyr Trp Thr Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Phe Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Thr Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Asp Thr Met Thr Gly Leu Asp Val Trp Gly Gln Gly

100 105 110

Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

130 135 140

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

145 150 155 160

Gln Ser Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys

165 170 175

Ala Pro Thr Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val

180 185 190

Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr

195 200 205

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Ala Tyr Phe Cys Gln Gln

210 215 220

Thr Tyr Ser Asn Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Val Lys

225 230 235 240

<210> 405

<211> 926

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1xADT1-4-2

<400> 405

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val Gln

210 215 220

Ser Gly Val Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys

225 230 235 240

Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Tyr Met Tyr Trp Val Arg

245 250 255

Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Gly Ile Asn Pro Ser

260 265 270

Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe Lys Asn Arg Val Thr Leu

275 280 285

Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr Met Glu Leu Lys Ser Leu

290 295 300

Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Arg Asp Tyr Arg

305 310 315 320

Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

325 330 335

Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser

340 345 350

Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys

355 360 365

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu

370 375 380

Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu

385 390 395 400

Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr

405 410 415

Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val

420 425 430

Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro

435 440 445

Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe

450 455 460

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

465 470 475 480

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe

485 490 495

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

500 505 510

Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

515 520 525

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

530 535 540

Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala

545 550 555 560

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg

565 570 575

Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

580 585 590

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

595 600 605

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

610 615 620

Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln

625 630 635 640

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

645 650 655

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Gly

660 665 670

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln

675 680 685

Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr

690 695 700

Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn

705 710 715 720

Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr

725 730 735

Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser

740 745 750

Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln

755 760 765

Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

770 775 780

Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr

785 790 795 800

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

805 810 815

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

820 825 830

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

835 840 845

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

850 855 860

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe

865 870 875 880

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

885 890 895

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala

900 905 910

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

915 920 925

<210> 406

<211> 246

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-PD-1 scFv binding Module

<400> 406

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

115 120 125

Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala

130 135 140

Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala

145 150 155 160

Ser Lys Gly Val Ser Thr Ser Gly Tyr Ser Tyr Leu His Trp Tyr Gln

165 170 175

Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Leu Ala Ser Tyr

180 185 190

Leu Glu Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr

195 200 205

Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val

210 215 220

Tyr Tyr Cys Gln His Ser Arg Asp Leu Pro Leu Thr Phe Gly Gly Gly

225 230 235 240

Thr Lys Val Glu Ile Lys

245

<210> 407

<211> 918

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2x 4-1BB

<400> 407

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly

660 665 670

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln Ser Gly Ala

675 680 685

Glu Val Lys Lys Pro Gly Glu Ser Leu Arg Ile Ser Cys Lys Gly Ser

690 695 700

Gly Tyr Ser Phe Ser Thr Tyr Trp Ile Ser Trp Val Arg Gln Met Pro

705 710 715 720

Gly Lys Gly Leu Glu Trp Met Gly Lys Ile Tyr Pro Gly Asp Ser Tyr

725 730 735

Thr Asn Tyr Ser Pro Ser Phe Gln Gly Gln Val Thr Ile Ser Ala Asp

740 745 750

Lys Ser Ile Ser Thr Ala Tyr Leu Gln Trp Ser Ser Leu Lys Ala Ser

755 760 765

Asp Thr Ala Met Tyr Tyr Cys Ala Arg Gly Tyr Gly Ile Phe Asp Tyr

770 775 780

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser

785 790 795 800

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Tyr Glu Leu Thr Gln

805 810 815

Pro Pro Ser Val Ser Val Ser Pro Gly Gln Thr Ala Ser Ile Thr Cys

820 825 830

Ser Gly Asp Asn Ile Gly Asp Gln Tyr Ala His Trp Tyr Gln Gln Lys

835 840 845

Pro Gly Gln Ser Pro Val Leu Val Ile Tyr Gln Asp Lys Asn Arg Pro

850 855 860

Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala

865 870 875 880

Thr Leu Thr Ile Ser Gly Thr Gln Ala Met Asp Glu Ala Asp Tyr Tyr

885 890 895

Cys Ala Thr Tyr Thr Gly Phe Gly Ser Leu Ala Val Phe Gly Gly Gly

900 905 910

Thr Lys Leu Thr Val Leu

915

<210> 408

<211> 239

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-41 BB scFv binding Module

<400> 408

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Ser Thr Tyr

20 25 30

Trp Ile Ser Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Lys Ile Tyr Pro Gly Asp Ser Tyr Thr Asn Tyr Ser Pro Ser Phe

50 55 60

Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Gly Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

115 120 125

Gly Gly Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser

130 135 140

Pro Gly Gln Thr Ala Ser Ile Thr Cys Ser Gly Asp Asn Ile Gly Asp

145 150 155 160

Gln Tyr Ala His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu

165 170 175

Val Ile Tyr Gln Asp Lys Asn Arg Pro Ser Gly Ile Pro Glu Arg Phe

180 185 190

Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr

195 200 205

Gln Ala Met Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Tyr Thr Gly Phe

210 215 220

Gly Ser Leu Ala Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

225 230 235

<210> 409

<211> 918

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2xOX40

<400> 409

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Gln Gln Ser Gly Pro

210 215 220

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

225 230 235 240

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

245 250 255

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

260 265 270

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

275 280 285

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

290 295 300

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

305 310 315 320

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

325 330 335

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

340 345 350

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

355 360 365

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

370 375 380

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

385 390 395 400

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

405 410 415

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

420 425 430

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

435 440 445

Ala Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

545 550 555 560

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

565 570 575

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

610 615 620

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly

660 665 670

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln Ser Gly Ala

675 680 685

Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser

690 695 700

Gly Tyr Thr Phe Thr Asp Ser Tyr Met Ser Trp Val Arg Gln Ala Pro

705 710 715 720

Gly Gln Gly Leu Glu Trp Ile Gly Asp Met Tyr Pro Asp Asn Gly Asp

725 730 735

Ser Ser Tyr Asn Gln Lys Phe Arg Glu Arg Val Thr Ile Thr Arg Asp

740 745 750

Thr Ser Thr Ser Thr Ala Tyr Leu Glu Leu Ser Ser Leu Arg Ser Glu

755 760 765

Asp Thr Ala Val Tyr Tyr Cys Val Leu Ala Pro Arg Trp Tyr Phe Ser

770 775 780

Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

785 790 795 800

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr

805 810 815

Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile

820 825 830

Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr Leu Asn Trp Tyr Gln

835 840 845

Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Arg

850 855 860

Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr

865 870 875 880

Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr

885 890 895

Tyr Tyr Cys Gln Gln Gly His Thr Leu Pro Pro Thr Phe Gly Gln Gly

900 905 910

Thr Lys Val Glu Ile Lys

915

<210> 410

<211> 239

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-OX 40 scFv binding modules

<400> 410

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Ser

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asp Met Tyr Pro Asp Asn Gly Asp Ser Ser Tyr Asn Gln Lys Phe

50 55 60

Arg Glu Arg Val Thr Ile Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Leu Ala Pro Arg Trp Tyr Phe Ser Val Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

130 135 140

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp

145 150 155 160

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

165 170 175

Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser

180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

195 200 205

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly His

210 215 220

Thr Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

225 230 235

<210> 411

<211> 934

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT x ADT1-4-2

<400> 411

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Lys Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Asn Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Glu Val Gln Leu

210 215 220

Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu

225 230 235 240

Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn Ser Ala Ala Trp

245 250 255

Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Lys

260 265 270

Thr Tyr Tyr Arg Phe Lys Trp Tyr Ser Asp Tyr Ala Val Ser Val Lys

275 280 285

Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn Gln Phe Ser Leu

290 295 300

Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val Phe Tyr Cys Thr

305 310 315 320

Arg Glu Ser Thr Thr Tyr Asp Leu Leu Ala Gly Pro Phe Asp Tyr Trp

325 330 335

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

340 345 350

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

355 360 365

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

370 375 380

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

385 390 395 400

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

405 410 415

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

420 425 430

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

435 440 445

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala

450 455 460

Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

465 470 475 480

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

485 490 495

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

500 505 510

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

515 520 525

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

530 535 540

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

545 550 555 560

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

565 570 575

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

580 585 590

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

595 600 605

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

610 615 620

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

625 630 635 640

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

645 650 655

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

660 665 670

Ser Pro Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

675 680 685

Gly Gly Ser Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys

690 695 700

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val

705 710 715 720

Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg

725 730 735

Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr

740 745 750

Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr

755 760 765

Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp

770 775 780

Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg

785 790 795 800

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser

805 810 815

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln

820 825 830

Ser Pro Pro Ala Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

835 840 845

Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp Leu Ala Trp Tyr Gln His

850 855 860

Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu

865 870 875 880

Glu Ser Gly Val Pro Leu Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu

885 890 895

Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr

900 905 910

Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr Phe Gly Gln Gly

915 920 925

Thr Arg Leu Glu Ile Lys

930

<210> 412

<211> 255

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti TIGIT SCFV binding module

<400> 412

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Lys Thr Tyr Tyr Arg Phe Lys Trp Tyr Ser Asp Tyr Ala

50 55 60

Val Ser Val Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Phe Tyr Cys Thr Arg Glu Ser Thr Thr Tyr Asp Leu Leu Ala Gly Pro

100 105 110

Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

130 135 140

Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg

145 150 155 160

Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Val Leu Tyr Ser Ser Asn

165 170 175

Asn Lys Lys Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

180 185 190

Asn Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp

195 200 205

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

210 215 220

Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr

225 230 235 240

Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Glu Ile Lys

245 250 255

<210> 413

<211> 243

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Anti-VD 1 scFv binding module

<400> 413

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

115 120 125

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Pro

130 135 140

Ala Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

145 150 155 160

Ser Gln Asp Ile Asn Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly

165 170 175

Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly

180 185 190

Val Pro Leu Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu

195 200 205

Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln

210 215 220

Gln Lys Tyr Ser Ala Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu

225 230 235 240

Glu Ile Lys

<210> 414

<211> 215

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 LC

<400> 414

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 415

<211> 716

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC + FAPa scFv

<400> 415

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

465 470 475 480

Ser Val Lys Val Ser Cys Lys Thr Ser Arg Tyr Thr Phe Thr Glu Tyr

485 490 495

Thr Ile His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile

500 505 510

Gly Gly Ile Asn Pro Asn Asn Gly Ile Pro Asn Tyr Asn Gln Lys Phe

515 520 525

Lys Gly Arg Val Thr Ile Thr Val Asp Thr Ser Ala Ser Thr Ala Tyr

530 535 540

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

545 550 555 560

Ala Arg Arg Arg Ile Ala Tyr Gly Tyr Asp Glu Gly His Ala Met Asp

565 570 575

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr

595 600 605

Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile

610 615 620

Asn Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser Arg Asn Gln Lys Asn

625 630 635 640

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

645 650 655

Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser

660 665 670

Gly Ser Gly Phe Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

675 680 685

Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Phe Ser Tyr Pro

690 695 700

Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

705 710 715

<210> 416

<211> 704

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC + MSLN scFv

<400> 416

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

465 470 475 480

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Asn

485 490 495

Asn Tyr Tyr Trp Thr Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu

500 505 510

Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Phe Tyr Asn Pro Ser

515 520 525

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Thr Gln Phe

530 535 540

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

545 550 555 560

Cys Ala Arg Glu Asp Thr Met Thr Gly Leu Asp Val Trp Gly Gln Gly

565 570 575

Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

595 600 605

Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

610 615 620

Gln Ser Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys

625 630 635 640

Ala Pro Thr Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val

645 650 655

Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr

660 665 670

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Ala Tyr Phe Cys Gln Gln

675 680 685

Thr Tyr Ser Asn Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Val Lys

690 695 700

<210> 417

<211> 708

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 HC + ADT1-4-2 scFv

<400> 417

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

450 455 460

Ser Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser

465 470 475 480

Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser

485 490 495

Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu

500 505 510

Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr

515 520 525

Ala Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys

530 535 540

Asn Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala

545 550 555 560

Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly

565 570 575

Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro

595 600 605

Pro Ala Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg

610 615 620

Ala Ser Gln Asp Ile Asn Asp Trp Leu Ala Trp Tyr Gln His Lys Pro

625 630 635 640

Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser

645 650 655

Gly Val Pro Leu Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr

660 665 670

Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys

675 680 685

Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr Phe Gly Gln Gly Thr Arg

690 695 700

Leu Glu Ile Lys

705

<210> 418

<211> 703

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC + 4-1BB scFv

<400> 418

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

465 470 475 480

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Ser Thr Tyr

485 490 495

Trp Ile Ser Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

500 505 510

Gly Lys Ile Tyr Pro Gly Asp Ser Tyr Thr Asn Tyr Ser Pro Ser Phe

515 520 525

Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

530 535 540

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

545 550 555 560

Ala Arg Gly Tyr Gly Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

565 570 575

Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser

595 600 605

Pro Gly Gln Thr Ala Ser Ile Thr Cys Ser Gly Asp Asn Ile Gly Asp

610 615 620

Gln Tyr Ala His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu

625 630 635 640

Val Ile Tyr Gln Asp Lys Asn Arg Pro Ser Gly Ile Pro Glu Arg Phe

645 650 655

Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr

660 665 670

Gln Ala Met Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Tyr Thr Gly Phe

675 680 685

Gly Ser Leu Ala Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

690 695 700

<210> 419

<211> 703

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC + OX40 scFv

<400> 419

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

465 470 475 480

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Ser

485 490 495

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

500 505 510

Gly Asp Met Tyr Pro Asp Asn Gly Asp Ser Ser Tyr Asn Gln Lys Phe

515 520 525

Arg Glu Arg Val Thr Ile Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr

530 535 540

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

545 550 555 560

Val Leu Ala Pro Arg Trp Tyr Phe Ser Val Trp Gly Gln Gly Thr Leu

565 570 575

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser

595 600 605

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp

610 615 620

Ile Ser Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

625 630 635 640

Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser

645 650 655

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

660 665 670

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly His

675 680 685

Thr Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

690 695 700

<210> 420

<211> 714

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT HC + ADT1-4-2 scFv

<400> 420

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Lys Thr Tyr Tyr Arg Phe Lys Trp Tyr Ser Asp Tyr Ala

50 55 60

Val Ser Val Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Phe Tyr Cys Thr Arg Glu Ser Thr Thr Tyr Asp Leu Leu Ala Gly Pro

100 105 110

Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

115 120 125

Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr

130 135 140

Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro

145 150 155 160

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val

165 170 175

His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser

180 185 190

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile

195 200 205

Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val

210 215 220

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

225 230 235 240

Pro Glu Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

245 250 255

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

260 265 270

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

275 280 285

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

290 295 300

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

305 310 315 320

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

325 330 335

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

340 345 350

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr

355 360 365

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

370 375 380

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

385 390 395 400

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

405 410 415

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

420 425 430

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

435 440 445

Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly

450 455 460

Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln Gln Ser Gly Pro

465 470 475 480

Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser

485 490 495

Gly Asp Ser Val Ser Ser Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln

500 505 510

Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly Arg Thr Tyr Tyr Arg Ser

515 520 525

Lys Trp Ser Thr Asp Tyr Ala Ala Ser Val Lys Ser Arg Ile Thr Ile

530 535 540

Asn Pro Asp Thr Ser Lys Asn Gln Leu Ser Leu Gln Leu Asn Ser Val

545 550 555 560

Thr Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Trp Val Gly

565 570 575

Tyr Val Asp Arg Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

580 585 590

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile

595 600 605

Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly Asp Arg

610 615 620

Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp Leu Ala

625 630 635 640

Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp

645 650 655

Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly Ser Gly

660 665 670

Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp

675 680 685

Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln Val Thr

690 695 700

Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

705 710

<210> 421

<211> 217

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G04 LC

<400> 421

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn

20 25 30

Asp Trp Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Pro Gln Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr

100 105 110

Ala Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 422

<211> 714

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G04 HC + CD19 scFv

<400> 422

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ser Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

465 470 475 480

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr

485 490 495

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

500 505 510

Gly Gln Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe

515 520 525

Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr

530 535 540

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys

545 550 555 560

Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp

565 570 575

Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Leu Thr

595 600 605

Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile

610 615 620

Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu

625 630 635 640

Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

645 650 655

Asp Ala Ser Asn Leu Val Ser Gly Ile Pro Pro Arg Phe Ser Gly Ser

660 665 670

Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Lys Val

675 680 685

Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr Glu Asp Pro Trp Thr

690 695 700

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

705 710

<210> 423

<211> 216

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E07 LC

<400> 423

Ala Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

1 5 10 15

Val Gly Asp Arg Val Thr Ile Ala Cys Arg Ala Gly Gln Ser Ile Gly

20 25 30

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

35 40 45

Leu Ile Tyr Val Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

85 90 95

Leu Leu Thr Phe Gly Arg Gly Thr Lys Val Glu Ile Lys Arg Thr Ala

100 105 110

Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys

115 120 125

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg

130 135 140

Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn

145 150 155 160

Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser

165 170 175

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys

180 185 190

Leu Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr

195 200 205

Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 424

<211> 712

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E07 HC + CD19 scFv

<400> 424

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Ala Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val

450 455 460

Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val

465 470 475 480

Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met

485 490 495

Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Gln

500 505 510

Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly

515 520 525

Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gln

530 535 540

Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg

545 550 555 560

Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp

565 570 575

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Leu Thr Gln Ser

595 600 605

Pro Ala Ser Leu Ala Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys

610 615 620

Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu Asn Trp

625 630 635 640

Tyr Gln Gln Ile Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Asp Ala

645 650 655

Ser Asn Leu Val Ser Gly Ile Pro Pro Arg Phe Ser Gly Ser Gly Ser

660 665 670

Gly Thr Asp Phe Thr Leu Asn Ile His Pro Val Glu Lys Val Asp Ala

675 680 685

Ala Thr Tyr His Cys Gln Gln Ser Thr Glu Asp Pro Trp Thr Phe Gly

690 695 700

Gly Gly Thr Lys Leu Glu Ile Lys

705 710

<210> 425

<211> 216

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 LC

<400> 425

Ala Ser Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro

1 5 10 15

Gly Lys Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Gln

20 25 30

Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Met Leu Val

35 40 45

Ile Tyr Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser

50 55 60

Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu

65 70 75 80

Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser

85 90 95

Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

100 105 110

Pro Ala Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

115 120 125

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

130 135 140

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys

145 150 155 160

Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr

165 170 175

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

180 185 190

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

195 200 205

Thr Val Ala Pro Thr Glu Cys Ser

210 215

<210> 426

<211> 446

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 HC (LAGA) EGFR FS1-67

<400> 426

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Ser Pro Ile Glu Leu Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Thr Asp Asp Gly Pro Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Thr

370 375 380

Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp Arg

405 410 415

Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 427

<211> 447

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E07 HC (LAGA) EGFR FS1-67

<400> 427

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Ala Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Thr Asp Asp Gly Pro Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser Tyr Trp

405 410 415

Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 428

<211> 702

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> C08 HC + EGFR scFv

<400> 428

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Ser Pro Ile Glu Leu Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly

435 440 445

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln

450 455 460

Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu Ser

465 470 475 480

Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val His

485 490 495

Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val Ile

500 505 510

Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg Leu

515 520 525

Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met Asn

530 535 540

Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala Leu

545 550 555 560

Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val

565 570 575

Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

580 585 590

Gly Gly Ser Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val

595 600 605

Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile

610 615 620

Gly Thr Asn Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg

625 630 635 640

Leu Leu Ile Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg

645 650 655

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser

660 665 670

Val Glu Ser Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn

675 680 685

Trp Pro Thr Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

690 695 700

<210> 429

<211> 705

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G04 HC + EGFR scFv

<400> 429

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ser Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

450 455 460

Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln

465 470 475 480

Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr

485 490 495

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

500 505 510

Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr

515 520 525

Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe

530 535 540

Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala

545 550 555 560

Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly

565 570 575

Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly

580 585 590

Ser Gly Gly Gly Gly Ser Asp Ile Leu Leu Thr Gln Ser Pro Val Ile

595 600 605

Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg Ala Ser

610 615 620

Gln Ser Ile Gly Thr Asn Ile His Trp Tyr Gln Gln Arg Thr Asn Gly

625 630 635 640

Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile

645 650 655

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser

660 665 670

Ile Asn Ser Val Glu Ser Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln

675 680 685

Asn Asn Asn Trp Pro Thr Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu

690 695 700

Lys

705

<210> 430

<211> 703

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> E07 HC + EGFR scFv

<400> 430

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Asp Tyr Ala Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly

435 440 445

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val

450 455 460

Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu

465 470 475 480

Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val

485 490 495

His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val

500 505 510

Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg

515 520 525

Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met

530 535 540

Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala

545 550 555 560

Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu

565 570 575

Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

580 585 590

Gly Gly Gly Ser Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser

595 600 605

Val Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser

610 615 620

Ile Gly Thr Asn Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro

625 630 635 640

Arg Leu Leu Ile Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser

645 650 655

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn

660 665 670

Ser Val Glu Ser Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn

675 680 685

Asn Trp Pro Thr Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

690 695 700

<210> 431

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (LAGA) EGFR FS1-67

<400> 431

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp Asp Gly Pro Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser

405 410 415

Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 432

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (LAGA) EGFR LEE

<400> 432

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Glu Glu Gly Pro Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser

405 410 415

Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 433

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (LAGA) HER2 Fcab

<400> 433

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Phe Phe Thr Tyr Trp Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Arg Arg Arg Trp Thr Ala Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 434

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (wt) HER2 Fcab

<400> 434

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Phe Phe Thr Tyr Trp Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

405 410 415

Arg Arg Arg Trp Thr Ala Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 435

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (wt) EGFR FS1-67

<400> 435

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp Asp Gly Pro Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser

405 410 415

Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 436

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4-2 HC (wt) EGFR LEE

<400> 436

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Val Gly Tyr Val Asp Arg Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Leu Glu Glu Gly Pro Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser

405 410 415

Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 437

<211> 449

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> G04 HC (LAGA) EGFR FS1-67

<400> 437

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Lys

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Ser Thr Asp Tyr Ala

50 55 60

Ala Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Leu Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Thr Trp Ser Gly Tyr Val Asp Val Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

210 215 220

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Ala Gly Ala

225 230 235 240

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

245 250 255

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

260 265 270

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

275 280 285

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

290 295 300

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

305 310 315 320

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

325 330 335

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

340 345 350

Thr Leu Pro Pro Ser Arg Asp Glu Thr Asp Asp Gly Pro Val Ser Leu

355 360 365

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

370 375 380

Glu Ser Thr Tyr Gly Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ser

405 410 415

Tyr Trp Arg Trp Tyr Lys Gly Asn Val Phe Ser Cys Ser Val Met His

420 425 430

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

435 440 445

Gly

<210> 438

<211> 218

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 LC

<400> 438

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 439

<211> 220

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT LC

<400> 439

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Lys Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Asn Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 440

<211> 124

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 VH

<400> 440

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ser

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr

20 25 30

Trp Met Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Gln Ile Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 441

<211> 111

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 VL

<400> 441

Asp Ile Gln Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Asp Tyr Asp

20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gln Gln Ile Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu Val Ser Gly Ile Pro Pro

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His

65 70 75 80

Pro Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gln Gln Ser Thr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 442

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 HCDR1

<400> 442

Gly Tyr Ala Phe Ser Ser Tyr Trp

1 5

<210> 443

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 HCDR2

<400> 443

Ile Trp Pro Gly Asp Gly Asp Thr

1 5

<210> 444

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 HCDR3

<400> 444

Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp

1 5 10 15

Tyr

<210> 445

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 LCDR1

<400> 445

Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr

1 5 10

<210> 446

<211> 2

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 LCDR2

<400> 446

Asp Ala

1

<210> 447

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> CD19 LCDR3

<400> 447

Gln Gln Ser Thr Glu Asp Pro Trp Thr

1 5

<210> 448

<211> 118

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR VH

<400> 448

Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln

1 5 10 15

Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr

20 25 30

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr

50 55 60

Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe

65 70 75 80

Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser

115

<210> 449

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR VL

<400> 449

Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn

20 25 30

Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile

35 40 45

Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser

65 70 75 80

Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210> 450

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR HCDR1

<400> 450

Gly Phe Ser Leu Thr Asn Tyr Gly

1 5

<210> 451

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR HCDR2

<400> 451

Ile Trp Ser Gly Gly Asn Thr

1 5

<210> 452

<211> 13

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR HCDR3

<400> 452

Ala Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr

1 5 10

<210> 453

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR LCDR1

<400> 453

Arg Ala Ser Gln Ser Ile Gly Thr Asn Ile His

1 5 10

<210> 454

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR LCDR2

<400> 454

Tyr Ala Ser Glu Ser Ile Ser

1 5

<210> 455

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> EGFR LCDR3

<400> 455

Gln Gln Asn Asn Asn Trp Pro Thr Thr

1 5

<210> 456

<211> 124

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa VH

<400> 456

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Thr Ser Arg Tyr Thr Phe Thr Glu Tyr

20 25 30

Thr Ile His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile

35 40 45

Gly Gly Ile Asn Pro Asn Asn Gly Ile Pro Asn Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Val Asp Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Arg Ile Ala Tyr Gly Tyr Asp Glu Gly His Ala Met Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 457

<211> 113

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa VL

<400> 457

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Arg Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Phe Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Phe Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 458

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa HCDR1

<400> 458

Arg Tyr Thr Phe Thr Glu Tyr Thr

1 5

<210> 459

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa HCDR2

<400> 459

Ile Asn Pro Asn Asn Gly Ile Pro

1 5

<210> 460

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa HCDR3

<400> 460

Ala Arg Arg Arg Ile Ala Tyr Gly Tyr Asp Glu Gly His Ala Met Asp

1 5 10 15

Tyr

<210> 461

<211> 17

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa LCDR1

<400> 461

Lys Ser Ser Gln Ser Leu Leu Tyr Ser Arg Asn Gln Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 462

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa LCDR2

<400> 462

Trp Ala Ser Thr Arg Glu Ser

1 5

<210> 463

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> FAPa LCDR3

<400> 463

Gln Gln Tyr Phe Ser Tyr Pro Leu Thr

1 5

<210> 464

<211> 119

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN VH

<400> 464

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asn Asn

20 25 30

Asn Tyr Tyr Trp Thr Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Phe Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Thr Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Asp Thr Met Thr Gly Leu Asp Val Trp Gly Gln Gly

100 105 110

Thr Thr Val Thr Val Ser Ser

115

<210> 465

<211> 106

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN VL

<400> 465

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Ala Tyr Phe Cys Gln Gln Thr Tyr Ser Asn Pro Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Val Lys

100 105

<210> 466

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN HCDR1

<400> 466

Gly Gly Ser Ile Asn Asn Asn Asn Tyr Tyr

1 5 10

<210> 467

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN HCDR2

<400> 467

Ile Tyr Tyr Ser Gly Ser Thr

1 5

<210> 468

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN HCDR3

<400> 468

Ala Arg Glu Asp Thr Met Thr Gly Leu Asp Val

1 5 10

<210> 469

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN LCDR1

<400> 469

Gln Ser Ile Asn Asn Tyr

1 5

<210> 470

<211> 2

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN LCDR2

<400> 470

Ala Ala

1

<210> 471

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> MSLN LCDR3

<400> 471

Gln Gln Thr Tyr Ser Asn Pro Thr

1 5

<210> 472

<211> 120

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 VH

<400> 472

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 473

<211> 111

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 VL

<400> 473

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 474

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 HCDR1

<400> 474

Gly Tyr Thr Phe Thr Asn Tyr Tyr

1 5

<210> 475

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 HCDR2

<400> 475

Ile Asn Pro Ser Asn Gly Gly Thr

1 5

<210> 476

<211> 13

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 HCDR3

<400> 476

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr

1 5 10

<210> 477

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 LCDR1

<400> 477

Lys Gly Val Ser Thr Ser Gly Tyr Ser Tyr

1 5 10

<210> 478

<211> 2

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 LCDR2

<400> 478

Leu Ala

1

<210> 479

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> PD-1 LCDR3

<400> 479

Gln His Ser Arg Asp Leu Pro Leu Thr

1 5

<210> 480

<211> 116

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB VH

<400> 480

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Ser Thr Tyr

20 25 30

Trp Ile Ser Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Lys Ile Tyr Pro Gly Asp Ser Tyr Thr Asn Tyr Ser Pro Ser Phe

50 55 60

Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr

65 70 75 80

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Gly Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 481

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB VL

<400> 481

Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln Thr

1 5 10 15

Ala Ser Ile Thr Cys Ser Gly Asp Asn Ile Gly Asp Gln Tyr Ala His

20 25 30

Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu Val Ile Tyr Gln

35 40 45

Asp Lys Asn Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser Asn

50 55 60

Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Met Asp

65 70 75 80

Glu Ala Asp Tyr Tyr Cys Ala Thr Tyr Thr Gly Phe Gly Ser Leu Ala

85 90 95

Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 482

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB HCDR1

<400> 482

Gly Tyr Ser Phe Ser Thr Tyr Trp

1 5

<210> 483

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB HCDR2

<400> 483

Ile Tyr Pro Gly Asp Ser Tyr Thr

1 5

<210> 484

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB HCDR3

<400> 484

Ala Arg Gly Tyr Gly Ile Phe Asp Tyr

1 5

<210> 485

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB LCDR1

<400> 485

Ser Gly Asp Asn Ile Gly Asp Gln Tyr Ala His

1 5 10

<210> 486

<211> 7

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB LCDR2

<400> 486

Gln Asp Lys Asn Arg Pro Ser

1 5

<210> 487

<211> 11

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> 4-1BB LCDR3

<400> 487

Ala Thr Tyr Thr Gly Phe Gly Ser Leu Ala Val

1 5 10

<210> 488

<211> 117

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 VH

<400> 488

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Ser

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asp Met Tyr Pro Asp Asn Gly Asp Ser Ser Tyr Asn Gln Lys Phe

50 55 60

Arg Glu Arg Val Thr Ile Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Leu Ala Pro Arg Trp Tyr Phe Ser Val Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 489

<211> 107

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 VL

<400> 489

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Arg Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly His Thr Leu Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 490

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 HCDR1

<400> 490

Gly Tyr Thr Phe Thr Asp Ser Tyr

1 5

<210> 491

<211> 8

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 HCDR2

<400> 491

Met Tyr Pro Asp Asn Gly Asp Ser

1 5

<210> 492

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 HCDR3

<400> 492

Val Leu Ala Pro Arg Trp Tyr Phe Ser Val

1 5 10

<210> 493

<211> 6

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 LCDR1

<400> 493

Gln Asp Ile Ser Asn Tyr

1 5

<210> 494

<211> 2

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 LCDR2

<400> 494

Tyr Thr

1

<210> 495

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> OX40 LCDR3

<400> 495

Gln Gln Gly His Thr Leu Pro Pro Thr

1 5

<210> 496

<211> 126

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT VH

<400> 496

Glu Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu

35 40 45

Trp Leu Gly Lys Thr Tyr Tyr Arg Phe Lys Trp Tyr Ser Asp Tyr Ala

50 55 60

Val Ser Val Lys Gly Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Phe Tyr Cys Thr Arg Glu Ser Thr Thr Tyr Asp Leu Leu Ala Gly Pro

100 105 110

Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 497

<211> 113

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT VL

<400> 497

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr Val Leu Tyr Ser

20 25 30

Ser Asn Asn Lys Lys Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Asn Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 498

<211> 10

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT HCDR1

<400> 498

Gly Asp Ser Val Ser Ser Asn Ser Ala Ala

1 5 10

<210> 499

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT HCDR2

<400> 499

Thr Tyr Tyr Arg Phe Lys Trp Tyr Ser

1 5

<210> 500

<211> 16

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT HCDR3

<400> 500

Thr Arg Glu Ser Thr Thr Tyr Asp Leu Leu Ala Gly Pro Phe Asp Tyr

1 5 10 15

<210> 501

<211> 12

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT LCDR1

<400> 501

Gln Thr Val Leu Tyr Ser Ser Asn Asn Lys Lys Tyr

1 5 10

<210> 502

<211> 2

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT LCDR2

<400> 502

Trp Ala

1

<210> 503

<211> 9

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> TIGIT LCDR3

<400> 503

Gln Gln Tyr Tyr Ser Thr Pro Phe Thr

1 5

<210> 504

<211> 215

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> ADT1-4 modified light chain (S74L)

<400> 504

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Leu Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 505

<211> 215

<212> PRT

<213> Chile (Homo Sapiens)

<220>

<223> Light chain recovered from ADT1-4-2 (L74S)

<400> 505

Asp Ile Gln Met Thr Gln Ser Pro Pro Ala Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asp Trp

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Lys Tyr Ser Ala Pro Gln

85 90 95

Val Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

Claims (29)

1. An anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, comprising:

a heavy chain variable region comprising:

A VHCDR1, the VHCDR1 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO: 51. 52 and 130;

A VHCDR2 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:53 and 131; and

A VHCDR3 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO: 68. 55 to 67, 69 to 78 and 133 to 143; and/or

A light chain variable region comprising:

A VLCDR1, the VLCDR1 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:79 and 144;

A VLCDR2 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO:80 and 145; and

A VLCDR3 comprising or consisting of an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a sequence selected from the group consisting of seq id nos: SEQ ID NO: 95. 82 to 94, 96 to 105 and 147 to 157.

2. The anti-vδ1 antibody or antigen-binding fragment thereof of claim 1, said anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a. comprising SEQ ID NOs: 51. 53 and 68, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 95, VLCDR1, VLCDR2, and VLCDR3;

b. comprising SEQ ID NOs: 51. 53 and 55, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 82, VLCDR1, VLCDR2, and VLCDR3;

c. Comprising SEQ ID NOs: 51. 53 and 56, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 83, VLCDR1, VLCDR2, and VLCDR3;

d. comprising SEQ ID NOs: 51. 53 and 57, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 84, VLCDR1, VLCDR2, and VLCDR3;

e. comprising SEQ ID NOs: 51. 53 and 58, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 85, VLCDR1, VLCDR2, and VLCDR3;

f. comprising SEQ ID NOs: 51. 53 and 59, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 86, VLCDR1, VLCDR2, and VLCDR3;

g. comprising SEQ ID NOs: 51. 53 and 60, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 87, VLCDR1, VLCDR2, and VLCDR3;

h. comprising SEQ ID NOs: 52. 53 and 61, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 88, VLCDR1, VLCDR2, and VLCDR3;

i. comprising SEQ ID NOs: 51. 53 and 62, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 89, VLCDR1, VLCDR2, and VLCDR3;

j. comprising SEQ ID NOs: 51. 53 and 63, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 90, VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of seq id no;

k. comprising SEQ ID NOs: 51. 53 and 64, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 91, VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of seq id nos;

1. comprising SEQ ID NOs: 51. 53 and 65, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 92, VLCDR1, VLCDR2, and VLCDR3;

m. comprising SEQ ID NO: 52. 53 and 66, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 93, VLCDR1, VLCDR2, and VLCDR3;

n. comprising SEQ ID NO: 51. 53 and 67, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 94, VLCDR1, VLCDR2, and VLCDR3;

o. comprising SEQ ID NO: 51. 53 and 69, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 96, VLCDR1, VLCDR2, and VLCDR3;

p. comprising SEQ ID NO: 51. 53 and 70, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 97, VLCDR1, VLCDR2, and VLCDR3;

q. comprises the amino acid sequence of SEQ ID NO: 51. 53 and 71, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 98, VLCDR1, VLCDR2, and VLCDR3;

r. comprising SEQ ID NO: 51. 53 and 72, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 99, VLCDR1, VLCDR2, and VLCDR3;

s. comprising SEQ ID NO: 51. 53 and 73, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 100, VLCDR1, VLCDR2, and VLCDR3;

t. comprises the amino acid sequence of SEQ ID NO: 52. 53 and 74, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 101, VLCDR1, VLCDR2, and VLCDR3;

u. comprising SEQ ID NO: 51. 53 and 75, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 102, VLCDR1, VLCDR2, and VLCDR3;

v. comprising SEQ ID NOs: 51. 53 and 76, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 103, VLCDR1, VLCDR2, and VLCDR3;

w. comprises SEQ ID NO: 51. 53 and 77, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 104, VLCDR1, VLCDR2, and VLCDR3;

x. comprises SEQ ID NOs: 51. 53 and 78, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 79. 80 and 105, VLCDR1, VLCDR2, and VLCDR3;

y. comprise the sequences of SEQ ID NOs: 130. 131 and 133, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 147, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of 145 and 147;

z. comprise the sequences of SEQ ID NOs: 130. 131 and 134, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 148, VLCDR1, VLCDR2, and VLCDR3;

aa. comprise the sequences of SEQ ID NOs: 130. 131 and 135, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 149, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of 145 and 149;

bb. comprise the sequences of SEQ ID NOs: 130. 131 and 136, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 150, VLCDR1, VLCDR2, and VLCDR3;

cc. comprise the sequences of SEQ ID NOs: 130. 131 and 137, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 151, and VLCDR1, VLCDR2, and VLCDR3;

dd. comprise the sequences of SEQ ID NOs: 130. 131 and 138, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 152, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of 145 and 152;

ee. comprise the sequences of SEQ ID NOs: 130. 131 and 139, and VHCDR1, VHCDR2 and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 153, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of 145 and 153;

ff. comprise the sequences of SEQ ID NOs: 130. 131 and 140, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 154, VLCDR1, VLCDR2, and VLCDR3;

gg. comprise the sequences of SEQ ID NOs: 130. 131 and 141, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 155, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of seq id nos;

hh. comprise the sequences of SEQ ID NOs: 130. 131 and 142, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 156, and VLCDR1, VLCDR2, and VLCDR3 of the amino acid sequences of 145 and 156; or (b)

Respectively comprising SEQ ID NO: 130. 131 and 143, and VHCDR1, VHCDR2, and VHCDR3 comprising the amino acid sequences of SEQ ID NOs: 144. 145 and 157, and VLCDR1, VLCDR2, and VLCDR3.

3. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof as claimed in claim 1 or claim 2, comprising:

A heavy chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 15. 2 to 14, 16 to 25 and 107 to 117 has or consists of a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical; and/or

A light chain variable region comprising a sequence selected from the group consisting of SEQ ID NOs: 40. 27 to 39, 41 to 50 and 119 to 129 has or consists of a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

4. The anti-vδ1 antibody or antigen-binding fragment thereof of any one of the previous claims, said anti-vδ1 antibody or antigen-binding fragment thereof comprising:

a. comprising a sequence identical to SEQ ID NO:15 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:40, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

b. Comprising a sequence identical to SEQ ID NO:2 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:27, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

c. Comprising a sequence identical to SEQ ID NO:3 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:28, VL having an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

d. Comprising a sequence identical to SEQ ID NO:4 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:29, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

e. Comprising a nucleotide sequence identical to seq id NO:5 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:30, a VL having an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

f. comprising a sequence identical to SEQ ID NO:6 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:31 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

g. comprising a sequence identical to SEQ ID NO:7 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:32, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

h. comprising a sequence identical to SEQ ID NO:8 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:33, VL having an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

i. Comprising a sequence identical to SEQ ID NO:9 and VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:34, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

j. Comprising a sequence identical to SEQ ID NO:10 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:35, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

k. Comprising a sequence identical to SEQ ID NO:11 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:36, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

1. comprising a sequence identical to SEQ ID NO:12 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:37 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

m. comprises a sequence identical to SEQ ID NO:13 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:38, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

n. comprises a sequence identical to SEQ ID NO:14 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:39 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

o. comprising a sequence identical to SEQ ID NO:16 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:41 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

p. comprising a sequence identical to SEQ ID NO:17 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:42 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

q. comprises a sequence identical to SEQ ID NO:18 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:43 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

r. comprises a sequence identical to SEQ ID NO:19 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:44 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

s. comprises a sequence identical to SEQ ID NO:20 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:45 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

t. comprises a sequence identical to SEQ ID NO:21 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:46 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

u. comprises a sequence identical to SEQ ID NO:22 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:47 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

v. comprising a sequence identical to SEQ ID NO:23 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:48 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

w. comprises a sequence identical to SEQ ID NO:24 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:49 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

x. comprises a sequence identical to SEQ ID NO:25 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:50 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

y. comprises a sequence identical to SEQ ID NO:107 and VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:119, VL having an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

z. comprises a sequence identical to SEQ ID NO:108 and VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:120, VL having an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

aa. comprises a sequence identical to SEQ ID NO:109 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:121, VL having an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

bb. comprises a sequence identical to SEQ ID NO:110 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:122 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

cc. comprises a sequence identical to SEQ ID NO:111 and VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:123 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

dd. comprises a sequence identical to SEQ ID NO:112 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:124 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

ee. comprises a sequence identical to SEQ ID NO:113 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:125 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

ff. comprises a sequence identical to SEQ ID NO:114 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:126 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity;

gg. comprises a sequence identical to SEQ ID NO:115 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:127 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity;

hh. comprises a sequence identical to SEQ ID NO:116 and VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:128 has an amino acid sequence of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity; or (b)

Comprising a sequence identical to SEQ ID NO:117 and a VH comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:129 has an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity.

5. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein said antibody or antigen binding fragment thereof comprises an amino acid residue other than serine at position 74 according to the IMGT kappa variable light chain sequence numbering system, optionally wherein said residue at position 74 is a leucine residue according to the IMGT numbering system.

6. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises:

an HCDR1 sequence comprising the sequence of GDSVSSKSX ₁ A (SEQ ID NO: 158);

an HCDR2 sequence comprising SEQ ID NO:53 sequence

An HCDR3 sequence comprising the sequence of X ₂WX₃X₄X₅X₆DX₇ (SEQ ID NO: 162), wherein the HCDR3 sequence is not SEQ ID NO:54;

an LCDR1 sequence comprising SEQ ID NO: 79;

an LCDR2 sequence comprising SEQ ID NO: 80; and

An LCDR3 sequence comprising a sequence of QQX ₈YX₉X₁₀X₁₁X₁₂X₁₃ T (SEQ ID NO: 166), wherein the LCDR3 sequence is not SEQ ID NO:81;

Wherein each of X ₁ to X ₁₃ is a naturally occurring amino acid,

Or wherein the antibody or antigen binding fragment thereof comprises:

an HCDR1 sequence comprising SEQ ID NO:130 sequence of 130

An HCDR2 sequence comprising SEQ ID NO:131 sequence

An HCDR3 sequence comprising the sequence of X ₁X₂YX₃X₄ AFDI wherein the HCDR3 sequence is not SEQ ID NO:132, a part of the material;

An LCDR1 sequence comprising SEQ ID NO:144, a sequence of seq id no;

an LCDR2 sequence comprising SEQ ID NO: 145; and

An LCDR3 sequence comprising a sequence of QQX ₅X₆X₇X₈LX₉ T wherein the LCDR3 sequence is not SEQ ID NO:146;

wherein each of X ₁ to X ₉ is a naturally occurring amino acid.

7. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, wherein the anti-vδ1 antibody or antigen-binding fragment thereof is an affinity matured variant of a parent anti-V61 antibody or antigen-binding fragment thereof, wherein the parent anti-V61 antibody or antigen-binding fragment thereof comprises:

a. Comprising SEQ ID NO:1 and a VH comprising the amino acid sequence of SEQ ID NO:26, a VL sequence of the amino acid sequence of seq id no;

b. comprising SEQ ID NO:106 and a VH comprising the amino acid sequence of SEQ ID NO:118, VL of the amino acid sequence of seq id no;

c. comprising SEQ ID NO:273 and VH comprising the amino acid sequence of SEQ ID NO:282, VL of the amino acid sequence of seq id no;

d. Comprising SEQ ID NO:274 and VH comprising the amino acid sequence of SEQ ID NO:283, VL of amino acid sequence;

e. comprising SEQ ID NO:275 and VH comprising the amino acid sequence of SEQ ID NO:284, VL of the amino acid sequence of seq id no;

f. comprising SEQ ID NO:276 and VH comprising the amino acid sequence of SEQ ID NO:285, VL of the amino acid sequence of 285;

g. comprising SEQ ID NO:277 and VH comprising the amino acid sequence of SEQ ID NO: 286;

h. Comprising SEQ ID NO:278 and VH comprising the amino acid sequence of SEQ ID NO: VL of amino acid sequence 287;

i. comprising SEQ ID NO:279 and VH comprising the amino acid sequence of SEQ ID NO:288, VL of the amino acid sequence;

j. comprising SEQ ID NO:280 and a VH comprising the amino acid sequence of SEQ ID NO:289, VL of an amino acid sequence;

k. Comprising SEQ ID NO:281 and VH comprising the amino acid sequence of SEQ ID NO: VL of the amino acid sequence 290; or (b)

L. comprising SEQ ID NO:312 and VH comprising the amino acid sequence of SEQ ID NO:313, VL of the amino acid sequence of seq id no;

Optionally wherein:

the affinity matured antibody has at least 20%, at least 30%, at least 40%, at least 50%, at least 100%, at least 500% or more preferably at least about 1000% higher affinity for binding to the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR) than the parent antibody, e.g. as measured by Kd;

The affinity-matured anti-vδ1 antibody or antigen-binding fragment thereof comprises VH and VL sequences that are at least 80%, at least 90%, at least 95% or at least 96% identical to the corresponding parent VH and VL sequences; and/or

The affinity matured anti-vδ1 antibody or antigen binding fragment comprises up to 20, such as up to 15, such as up to 10 amino acid substitutions compared to the parent antibody sequence.

8. An anti-vδ1 antibody or antigen binding fragment, wherein the antibody is a multispecific antibody that specifically binds to an epitope of:

a. a first target epitope, wherein the first target epitope is an epitope of the variable δ1 (vδ1) chain of γδ T Cell Receptor (TCR); and

B. A second target epitope;

Wherein the multispecific antibody comprises SEQ ID NO：385;SEQ ID NO：386;SEQ ID NO：387;SEQ ID NO：314;SEQ ID NO：393;SEQ ID NO：394;SEQ ID NO：395;SEQ ID NO：397;SEQ ID NO：388;SEQ ID NO：315;SEQ ID NO：316;SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：393;SEQ ID NO：396;SEQ ID NO：398;SEQ ID NO：399;SEQ ID NO：401;SEQ ID NO：402;SEQ ID NO：403;SEQ ID NO：404;SEQ ID NO：405;SEQ ID NO：406;SEQ ID NO：407;SEQ ID NO：408;SEQ ID NO：409;SEQ ID NO：410;SEQ ID NO：411;SEQ ID NO：412;SEQ ID NO：413;SEQ ID NO：414 and SEQ ID NO:415, a base; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:418; SEQ ID NO:505 and SEQ ID NO:415, a base; SEQ ID NO:505 and SEQ ID NO:416; SEQ ID NO:505 and SEQ ID NO:419; SEQ ID NO:505 and SEQ ID NO:418; SEQ ID NO:421 and SEQ ID NO:422, a part of the material; SEQ ID NO:504 and SEQ ID NO:422, a part of the material; SEQ ID NO:423 and SEQ ID NO: 424. SEQ ID NO:425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:429; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:414 and SEQ ID NO:435; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:434, a base; SEQ ID NO:505 and SEQ ID NO:435; SEQ ID NO:438 and SEQ ID NO:417; or SEQ ID NO:439 and SEQ ID NO:420.

9. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof, the anti-vδ1 antibody or antigen-binding fragment thereof characterized in that it:

a. Causing the TCR on vδ1T cells to which it binds to down-regulate;

b. does not exhibit CDC or ADCC; and

C. The vδ1t cells were not depleted.

10. The anti-TCR delta variable 1 (anti-vδ1) antibody, or antigen-binding fragment thereof, of any one of the preceding claims, wherein the multispecific antibody causes less than about 30%, or less than about 20%, or less than about 10% of a population of viable vδ T + cells to be depleted via ADCC and/or CDC.

11. The anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof of any one of the preceding claims, wherein the antibody or antigen-binding fragment thereof binds to human TRVD1 with a KD of less than about 10nM, optionally wherein the antibody or antigen-binding fragment thereof binds to cynomolgus monkey TRVD1 with a KD of less than about 100 nM.

12. An anti-tcrδ variable 1 (anti-vδ1) antibody or antigen binding fragment thereof, the anti-vδ1 antibody or antigen binding fragment thereof specifically binding to a variable δ1 (vδ1) chain of a human γδ T Cell Receptor (TCR) with a KD of less than about 10nM, optionally wherein the anti-vδ1 antibody or antigen binding fragment thereof binds to a variable δ1 (vδ1) chain of a cynomolgus monkey γδ T Cell Receptor (TCR) with a KD of less than about 100 nM.

13. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein the antibody or antigen binding fragment thereof:

a. Binding to human TRDV1 (SEQ ID NO 272 or 306) with a binding affinity (K _D, e.g., as measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

b. Optionally binds to cynomolgus monkey TRDV1 (SEQ ID NO 308) with a binding affinity (K _D, e.g. measured by surface plasmon resonance) of less than about 100nM, preferably less than about 50 nM;

c. the IC50 for TCR downregulation is less than about 50nM (preferably less than about 10 nM);

d. the IC50 for killing THP-1 cells is less than about 10nM (preferably less than about 5 nM); and

E. the IC90 for killing THP-1 cells is less than about 50nM.

14. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein said antibody or antigen binding fragment thereof binds only to SEQ ID NO: 272. SEQ ID NO:306 and/or SEQ ID NO:308 amino acid residues 1-90.

15. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein said antibody or antigen binding fragment thereof binds to an epitope within the following amino acid region:

(i) SEQ ID NO: 272. SEQ ID NO:306 and/or SEQ ID NO:308 of 3-20; and/or

(Ii) SEQ ID NO: 272. SEQ ID NO:306 and/or SEQ ID NO:308, 37-77.

16. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein the epitope is an activating epitope of γδ T cells, optionally wherein binding of the activating epitope: (i) down-regulating the γδ TCR; (ii) activating degranulation of said γδ T cells; and/or (iii) promote γδ T cell mediated killing.

17. The anti-vδ1 antibody or antigen binding fragment thereof of claim 16, wherein binding of said activating epitope upregulates expression of CD107a, CD25, CD69 and/or Ki 67.

18. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein the antibody is an IgG antibody, optionally wherein the antibody is an IgG1 antibody.

19. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein the antibody is a multispecific antibody that specifically binds to a variable δ1 (vδ1) chain of a γδ T Cell Receptor (TCR) and a second antigen, wherein the second antigen is a cancer antigen or a cancer-associated antigen, an immunomodulatory antigen, or a cluster of differentiation CD antigen.

20. The anti-vδ1 antibody or antigen binding fragment thereof of claim 19, wherein:

The cancer antigen or cancer-associated antigen is selected from the group consisting of: AFP, AKAP-4, ALK, alpha fetoprotein, androgen receptor, B7H3, BAGE, BCA225, BCA, bcr-abl, beta-catenin, beta-HCG, beta-human chorionic gonadotrophin, BORIS, BTAA, CA, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, CAM 17.1, CAM43, carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33/IL3Ra, CD68\P1, CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG 4), C-Met, CO-029, CSPG4, cyclin B1, cyclophilin C-related protein, CYP1B1, E2A-PRL, EGFR, EGFRvIII, ELF2M, epCAM, ephA, ephran B2, ephran-Bar virus antigen EBVA, ERG (TMPRSS 2ETS fusion gene) ETV6-AML, FAP, FGF-5, fos associated antigen 1, fucosyl GM1, G250, ga733\EpCAM, GAGE-1, GAGE-2, GD3, glioma associated antigen, globoH, glycolipid F77, GM3, GP 100 (Pmel 17), H4-RET, HER-2/Neu/ErbB-2, high molecular weight melanoma associated antigen (HMW-MAA) HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin Growth Factor (IGF) -I, intestinal carboxylesterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, legumain, LMP2, M344, MA-50, mac-2 binding protein 、MAD-CT—1、MAD-CT-2、MAGE、MAGE A1、MAGE A3、MAGE-1、MAGE-3、MAGE-4、MAGE-5、MAGE-6、MART-1、MART-1/MelanA、M-CSF、 melanoma-associated chondroitin sulfate proteoglycan (MCSP), mesothelin, MG7-Ag, ML-IAP, MN-CA IX, MOV18, MUC1, mum-1, hsp70-2, MYCN, MYL-RAR, NA17, NB/70K, neuronal-glial antigen 2 (NG 2), neutrophil elastase, nm-23H1, nuMa, NY-BR-1, NY-CO-1, NY-ESO-1, OY-TES1, p15, p16, p180erbB3, p185erbB2, p53 mutant, page4, PAX3, PAX5, PDGFR-beta, PLAC1, polysialic acid, POLYSAC prostate cancer tumor antigen-1 (PCTA-1), prostate specific antigen, prostatic Acid Phosphatase (PAP), protease 3 (PR 1), PSA, PSCA, PSMA, RAGE-1, ras mutant, RCAS1, RGS5, rhoC, ROR1, RU2 (AS), SART3, SDCCAG, sLe (a), sperm protein 17, SSX2, STn, survivin, TA-90, TAAL, TAG-72, telomerase, thyroglobulin, tie 2, TLP, tn, TPS, TRP-1, TRP-2, TSP-180, tyrosinase, VEGF, VEGFR2, VISTA, WT1, XAGE 1, 43-9F, 5T4, and 791Tgp72;

The immunomodulatory antigen is selected from the group consisting of ：B7-1(CD80)、B7-2(CD86)、B7-DC(CD273)、B7-H1(CD274)、B7-H2(CD275)、B7-H3(CD276)、B7-H4(VTCN1)、B7-H5(VISTA)、BTLA(CD272)、CD137、CD137L、CD24、CD27、CD28、CD38、CD40、CD40L(CD154)、CD54、CD59、CD70、CTLA4(CD152)、CXCL9、GITR(CD357)、HVEM(CD270)、ICAM-1(CD54)、ICOS(CD278)、LAG-3(CD223)、OX40(CD134)、OX40L(CD252)、PD-1(CD279)、PD-L1(CD274)、TIGIT、CD314、CD334、CD335、CD337 and TIM-3 (CD 366); or (b)

The CD antigen is selected from the group ：CD1a、CD1b、CD1c、CD1d、CD1e、CD2、CD3、CD3d、CD3e、CD3g、CD4、CD5、CD6、CD7、CD8、CD8a、CD8b、CD9、CD10、CD11a、CD11b、CD11c、CD11d、CD13、CD14、CD15、CD16、CD16a、CD16b、CD17、CD18、CD19、CD20、CD21、CD22、CD23、CD24、CD25、CD26、CD27、CD28、CD29、CD30、CD31、CD32A、CD32B、CD33、CD34、CD35、CD36、CD37、CD38、CD39、CD40、CD41、CD42、CD42a、CD42b、CD42c、CD42d、CD43、CD44、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD50、CD51、CD52、CD53、CD54、CD55、CD56、CD57、CD58、CD59、CD60a、CD60b、CD60c、CD61、CD62E、CD62L、CD62P、CD63、CD64a、CD65、CD65s、CD66a、CD66b、CD66c、CD66d、CD66e、CD66f、CD68、CD69、CD70、CD71、CD72、CD73、CD74、CD75、CD75s、CD77、CD79A、CD79B、CD80、CD81、CD82、CD83、CD84、CD85A、CD85B、CD85C、CD85D、CD85F、CD85G、CD85H、CD85I、CD85J、CD85K、CD85M、CD86、CD87、CD88、CD89、CD90、CD91、CD92、CD93、CD94、CD95、CD96、CD97、CD98、CD99、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107、CD107a、CD107b、CD108、CD109、CD110、CD111、CD112、CD113、CD114、CD115、CD116、CD117、CD118、CD119、CD120、CD120a、CD120b、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD129、CD130、CD131、CD132、CD133、CD134、CD135、CD136、CD137、CD138、CD139、CD140A、CD140B、CD141、CD142、CD143、CD144、CDw145、CD146、CD147、CD148、CD150、CD151、CD152、CD153、CD154、CD155、CD156、CD156a、CD156b、CD156c、CD157、CD158、CD158A、CD158B1、CD158B2、CD158C、CD158D、CD158E1、CD158E2、CD158F1、CD158F2、CD158G、CD158H、CD158I、CD158J、CD158K、CD159a、CD159c、CD160、CD161、CD162、CD163、CD164、CD165、CD166、CD167a、CD167b、CD168、CD169、CD170、CD171、CD172a、CD172b、CD172g、CD173、CD174、CD175、CD175s、CD176、CD177、CD178、CD179a、CD179b、CD180、CD181、CD182、CD183、CD184、CD185、CD186、CD187、CD188、CD189、CD190、CD191、CD192、CD193、CD194、CD195、CD196、CD197、CDw198、CDw199、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD207、CD208、CD209、CD210、CDw210a、CDw210b、CD211、CD212、CD213a1、CD213a2、CD214、CD215、CD216、CD217、CD218a、CD218b、CD219、CD220、CD221、CD222、CD223、CD224、CD225、CD226、CD227、CD228、CD229、CD230、CD231、CD232、CD233、CD234、CD235a、CD235b、CD236、CD237、CD238、CD239、CD240CE、CD240D、CD241、CD242、CD243、CD244、CD245、CD246、CD247、CD248、CD249、CD250、CD251、CD252、CD253、CD254、CD255、CD256、CD257、CD258、CD259、CD260、CD261、CD262、CD263、CD264、CD265、CD266、CD267、CD268、CD269、CD270、CD271、CD272、CD273、CD274、CD275、CD276、CD277、CD278、CD279、CD280、CD281、CD282、CD283、CD284、CD285、CD286、CD287、CD288、CD289、CD290、CD291、CD292、CDw293、CD294、CD295、CD296、CD297、CD298、CD299、CD300A、CD300C、CD301、CD302、CD303、CD304、CD305、CD306、CD307、CD307a、CD307b、CD307c、CD307d、CD307e、CD308、CD309、CD310、CD311、CD312、CD313、CD314、CD315、CD316、CD317、CD318、CD319、CD320、CD321、CD322、CD323、CD324、CD325、CD326、CD327、CD328、CD329、CD330、CD331、CD332、CD333、CD334、CD335、CD336、CD337、CD338、CD339、CD340、CD344、CD349、CD351、CD352、CD353、CD354、CD355、CD357、CD358、CD360、CD361、CD362、CD363、CD364、CD365、CD366、CD367、CD368、CD369、CD370 and CD371 consisting of.

21. The anti-vδ1 antibody or antigen binding fragment thereof of any one of claims 1-18, wherein the antibody is not a bispecific antibody or a multispecific antibody.

22. The anti-vδ1 antibody or antigen binding fragment thereof of any one of claims 1-20, wherein the antibody is a bispecific antibody and wherein:

a. The second antigen is CD19, optionally wherein the antibody comprises SEQ ID NO:314, a step of; SEQ ID NO:315; SEQ ID NO:316, a step of; SEQ ID NO:396; SEQ ID NO:397; SEQ ID NO:421 and SEQ ID NO:422, a part of the material; SEQ ID NO:504 and SEQ ID NO:422, a part of the material; or SEQ ID NO:423 and SEQ ID NO: 424.

B. The second antigen is Her2 (CD 340), optionally wherein the antibody comprises the amino acid sequence of SEQ ID NO:393; SEQ ID NO:394; SEQ ID NO:395; SEQ ID NO:398; SEQ ID NO:414 and SEQ ID NO:433, respectively; SEQ ID NO:504 and SEQ ID NO:433, respectively; SEQ ID NO:505 and SEQ ID NO:433, respectively; SEQ ID NO:414 and SEQ ID NO:434, a base; SEQ ID NO:504 and SEQ ID NO:434, a base; or SEQ ID NO:505 and SEQ ID NO:434, a base;

c. The second antigen is EGFR, optionally wherein the antibody comprises SEQ ID NO：378;SEQ ID NO：379;SEQ ID NO：380;SEQ ID NO：382;SEQ ID NO：383;SEQ ID NO：384;SEQ ID NO：385;SEQ ID NO：386;SEQ ID NO：387;SEQ ID NO：388;SEQ ID NO：399;SEQ ID NO：425 and SEQ ID NO:426; SEQ ID NO:421 and SEQ ID NO:437; SEQ ID NO:504 and SEQ ID NO:437; SEQ ID NO:423 and SEQ ID NO:427, a step of forming a pattern; SEQ ID NO:425 and SEQ ID NO:428, 428 of the base material. SEQ ID NO:421 and SEQ ID NO:429; SEQ ID NO:504 and SEQ ID NO:429; SEQ ID NO:423 and SEQ ID NO:430; SEQ ID NO:414 and SEQ ID NO:431; SEQ ID NO:414 and SEQ ID NO:435; SEQ ID NO:504 and SEQ ID NO:431; SEQ ID NO:504 and SEQ ID NO:435; SEQ ID NO:505 and SEQ ID NO:431; or SEQ ID NO:505 and SEQ ID NO:435;

d. the second antigen is fapα, optionally wherein the antibody comprises SEQ ID NO:401; SEQ ID NO:402; SEQ ID NO:414 and SEQ ID NO:415, a base; SEQ ID NO:504 and SEQ ID NO:415, a base; or SEQ ID NO:505 and SEQ ID NO:415, a base;

e. the second antigen is MSLN, optionally wherein the antibody comprises SEQ ID NO:403; SEQ ID NO:404; SEQ ID NO:414 and SEQ ID NO:416; SEQ ID NO:504 and SEQ ID NO:416; or SEQ ID NO:505 and SEQ ID NO:416;

f. The second antigen is PD-1, optionally wherein the antibody comprises SEQ ID NO:405; SEQ ID NO:406; or SEQ ID NO:438 and SEQ ID NO:417;

g. The second antigen is 4-1BB, optionally wherein the antibody comprises SEQ ID NO:407, a step of selecting a specific code; SEQ ID NO:408; or SEQ ID NO:414 and SEQ ID NO:418; SEQ ID NO:504 and SEQ ID NO:418; or SEQ ID NO:505 and SEQ ID NO:418;

h. The second antigen is OX40, optionally wherein the antibody comprises SEQ ID NO:409; SEQ ID NO:410; SEQ ID NO:414 and SEQ ID NO:419; SEQ ID NO:504 and SEQ ID NO:419; or SEQ ID NO:504 and SEQ ID NO:419; or (b)

I. The second antigen is TIGIT, optionally wherein the antibody comprises SEQ ID NO:411; SEQ ID NO:412; or SEQ ID NO:439 and SEQ ID NO:420.

23. The anti-vδ1 antibody or antigen binding fragment thereof of any one of the previous claims, wherein the antibody is Fc-effective.

24. A pharmaceutical composition comprising the antibody or antibody-binding fragment thereof of any one of claims 1 to 22 and a pharmaceutically acceptable diluent or carrier.

25. A method of treating a disease or disorder in a subject or of modulating an immune response in a subject, the method comprising administering to the subject the anti-vδ1 antibody or antibody binding fragment of any one of claims 1-22 or the pharmaceutical composition of claim 24.

26. The method of claim 25, wherein the disease or disorder is cancer, infectious disease, or inflammatory disease, or wherein the subject who modulates an immune response has cancer, infectious disease, or inflammatory disease.

27. The method of claim 24, wherein modulating an immune response in a subject comprises at least one selected from the group consisting of: activating γδ T cells, causing or increasing γδ T cell proliferation, causing or increasing γδ T cell expansion, causing or increasing γδ T cell degranulation, causing or increasing γδ T cell mediated killing activity, causing or increasing γδ T cytotoxicity, causing or increasing γδ T cell mobilization, increasing γδ T cell survival, and increasing resistance to γδ T cell depletion.

28. The method of any one of claims 25 to 27, wherein diseased cells are killed and healthy cells are retained.

29. An anti-TCR delta variable 1 (anti-vδ1) antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 22, or a pharmaceutical composition as claimed in claim 24, for use in the treatment of cancer, infectious disease or inflammatory disease.