EP2111415A2

EP2111415A2 - Monoclonal agonistic anti-ctla-4 antibodies

Info

Publication number: EP2111415A2
Application number: EP05826461A
Authority: EP
Inventors: Thomas TeGenero AG HANKE; Frank TeGenero AG HORLING; Martin TeGenero AG TRISCHLER
Original assignee: TheraMAB GmbH
Current assignee: TheraMAB LLC
Priority date: 2004-12-23
Filing date: 2005-12-22
Publication date: 2009-10-28
Also published as: WO2006066568A3; DE102004063494A1; WO2006066568A2; US20090123477A1

Abstract

The invention relates to a CTLA-4-specific, agonistic, isolated monoclonal antibody which does not bond to the C''D loop of CTLA-4.

Description

antibody

Field of the invention

The invention relates to an antibody which is specific for CTLA-4, a pharmaceutical composition containing such an antibody, nucleic acids encoding such antibodies, vectors containing such antibodies, cells transfected with such vectors, uses of such antibodies, methods for producing such antibodies, and methods for Preparation of a pharmaceutical composition containing such antibodies.

Background of the invention and prior art

T lymphocytes (T cells) are the main agents of a highly efficient immune response that protects the human body against invading pathogens such as bacteria and viruses. They regulate the molecular interaction between different cellular components of the immune system such as dendritic cells, B cells, macrophages or other T cells and perform important effector functions such as the destruction of virus-infected cells or tumor cells themselves. As a result, they occupy a key position in the initiation and coordination of an immune response.

Highly selective molecules called T cell antigen receptors (TCRs), located on the cell surface, give each T cell its identity and endow it with the ability to specifically recognize antigens derived from Molecules of the major histocompatibility complex (HLA) are out. Additional cell-surface receptors of the "CD" nomenclature regulate the manner of the T-cell response initiated by antigen-dependent stimulation of the TCR, thus dictating the TCR

Specificity of an immune response, whereas the CD receptors control the extent and quality of the response of the T cell. Under physiological conditions, a signal combination of TCR and at least one other CD receptor is required for the complete activation of T cells, which is characterized in particular by proliferation and cytokine production. This process is called "costimulation." The most important costimulatory CD molecule on quiescent human T cells is the CD28 molecule.

In order to avoid an overreaction of the immune system, which would lead to an uncontrolled and thus dangerous multiplication of lymphocytes and a massive release of inflammation - promoting cytokines, the

Activation of T cells are effectively shut down. This task is accomplished through the interaction of a number of immunological control mechanisms. A particularly important role is played by inhibitory cell surface receptors such as the "Cytotoxic T-Lymphocyte Antigen-4" (CTLA-4) molecule, which will be explained in detail later.

In the development of autoimmune diseases such as rheumatoid arthritis, type I diabetes, multiple sclerosis, colitis, or psoriasis as well as in the development of allergies plays an uncontrolled response of T-lymphocytes to the body's own structures or external antigens important role . Thus, an initially excessive activation of T cells, a lack of inhibition of autoreactive T cells, or even a deficient number and / or function of regulatory T cells may be causally related to these diseases. Also in allogeneic

Organ transplants, d. H . in transplantation among non-HLA-identical individuals, activation of the T cells of the recipient is undesirable because activation of T cells by recognition of allo-antigens is the major cause of a chronic rejection reaction.

Current therapeutic approaches to suppress T-cell response targeting the antigen nonspecific suppression ^'of the activity of both harmful and beneficial T cells by "non-specific" immunosuppressants from. This therapeutic effects are often accompanied by serious side effects.

CTLA-4 ^' (CD152) is a member of the immunoglobulin superfamily and structurally the closest relative of CD28 (Lenschow DJ, Walunas TL, Bluestone JA, CD28 / B7 system of T cell costimulation., Annu Rev Immunol, 1996. 14: 233- 58). In contrast to CD28, the physiological function of CTLA-4 is not the promotion, but the inhibition of T-cell activation. CTLA-4 is very weak on quiescent T cells and strongly expressed on the cell surface of activated and regulatory T cells. The binding of CTLA-4 to its natural ligands B7-1 (CD80) and B7-2 (CD86), which are expressed by antigen-presenting cells (APC), leads to a shutdown of T cell proliferation and suppression of the Cytokine expression (Egen JG, Kuhns MS, Allison JP, CTLA-4: new entry into biological function and use in tumor immunotherapy, Nat Immunol, 2002. 3 (7): 611-8). The inhibitory function of CTLA-4 on the T cell surface was first determined using immobilized monoclonal antibodies specific for mouse CTLA-4 molecule (Walunas TL, Lenschow DJ, Bakker CY, Linsley PS, Freeman GJ, Green JM, Thompson CB, Bluestone JA _ ^ _ CTLA-4 can function as a negative regulator of cell activation, Immunity, 1994.1 (5): 405-13) and man (Blair PJ, Riley JL, Levine BL, Lee KP, Craighead N, Francomano T, Perfetto SJ, Gray GS, Carreno BM, June CH, CTLA-4 ligation dellvers a unique signal to resting human

CD4 T cells that interleukin-2 secretion but allows BCI-X (L) induction, J Immunol, 1998. 160 (1): 12-5), and could be demonstrated by the phenotype of mice receiving CTLA-4 Gene was specifically inactivated by homologous recombination. These animals rapidly die of lymphoproliferative disease characterized by uncontrolled activation of T cells (Tivol EA, Borriello F, Schweitzer AN, Lynch WP, Bluestone JA, Sharpe AH, Loss of CTLA-4 leads to massive lymphoproliferation and fatal Immunity, 1995. 3 (5): 541-7, as well as Waterhouse P, Penninger JM, Timms E, Wakeham A, Shahinian A, Lee KP, Thompson CB, Griesser H, Mak TW, Lymphoproliferative disorders with early lethality in mice deficient in Ctla-4, Science, 1995. 270 (5238): 985-8).

Conversely, these results suggest that CTLA-4 blockade enhances the activation of T cells in vivo. Accordingly, blocking, ie antagonistic, anti-CTLA-4 antibodies potentiated an anti-tumor response (Chambers CA, Allison JP, Co-stimulation in cell responses, Curr Opin Immunol, 1997. 9 (3): 396-404), to lead but also autoimmunity (Luhder F, Hoglund P, Allison JP, Benoist C, Mathis D, Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) regulates the unfolding of autoimmune diabetes J Exp Med, 1998. 187 (3)): 427-32). These findings, which were initially obtained in the mouse system, could be confirmed in humans in the first clinical trials. In isolated cases the administration of a blocking anti-human CTLA-4 antibody led to a (partial) remission in patients with metastatic melanoma (Hodi FS, Mihm MC, Soiffer RJ, Haluska FG, Butler M, Seiden MV, Davis T,

Henry Spiers R, MacRae S, Willman A, Padera R, Jaklitsch MT, Shankar S, Chen TC, Korman A, Allison JP, Dranoff G, Biology activi ty of cytotoxic T lymphocyte-associated antigen 4 antibody blockade in previously vaccinated metastatic melanoma and ovarian carcinoma equivalents, Proc Natl Acad. USA, 2003. 100 (8): 4712-7). At the same time, clinical signs of autoimmunity were found in a high proportion of treated patients (Phan GQ, Yang JC, Sherry RM, Hwu P, Topalian SL, Schwartzentruber DJ, Restifo NP, Haworth LR, Seipp CA, Freezer LJ, Morton KE, Mavroukakis SA, Duray PH, Steinberg SM, Allison JP, Davis TA, Rosenberg SA, Cancer regression and autoimmune induced by cytotoxic T lymphocyte-associated antigen 4 blockade in patients wi th metastatic melanoma, Proc Natl Acad Sei USA, 2003. 100 ( 14): 8372-7).

A polymorphism in the CTLA-4 gene that results in decreased expression and functionality of the CTLA-4 protein correlates with an increased likelihood of human involvement in the autoimmune diseases rheumatoid arthritis (Seidl C, Donner H, Fischer B, Usadel KH, Seifried E , Coldwater JP, Badenhoop K, CTLA4 codon 11 dimorphism in patients with rheumatoid arthritis Tissue Antigens, 1998. Jan; 51 (1): 62-6), multiple sclerosis (Harbo HF, Celius EG, Vartdal F, Spurkland A, CTLA4 promoter and exon 1 dimorphisms in multiple sclerosis, Tissue Antigens, 1999; 53 (1): 106-10) or Type I Diabetis (Donner H, Rau H, Walfish PG, Braun J, Siegmund T, Finke R, Herwig J, Usadel KH,

Badenhoop K, CTLA4 alanine-17 confers genetic susceptibility to Graves' disease and type 1 diabetes mellitus. J Clin Endocrinol Metab, 1997. 82 (1): 143-6).

In contrast to enhancing a T cell response with the anti-CTLA-4 blocking / antagonistic antibodies described above, agonist anti-CTLA-4 antibodies should be immunosuppressive. However, this has so far only been convincingly demonstrated for artificially immobilized antibodies. Thus, the genetically engineered transmembrane expression of an anti-CTLA-4 "single chain" antibody on artificial APC reduced TCR mediated proliferation and interleukin-2 secretion of T cells (Griffin MD, Hong DK, Holman PO, Lee KM, Whitters MJ, Mammin SM, Fallarino F, Collins M, Segal DM, Gaj ewski TF, Wreath DM, Bluestone JA, Blockade of T cell activation using a surface-linked single-chain antibody to CTLA-4 (CD152) .J Immunol, 2000. 164 (9): 4433-42). The fact that not only pre-activated but also quiescent T cells were inhibited in this experimental approach demonstrates that an important function of CTLA-4 is the early suppression of the TCR signal Similar results were reported by Brunner et al. (Brunner MC, Chambers CA, Chan FK, Hanke J, Winoto A, Allison JP, CTLA-4 mediated inhibition of early events of cell proliferation J Immunol, 1999, 162 (10): 5813-20) in the analysis of CTLA-4 signaling pathways in naive T cells. The transmembrane expression of an anti-CTLA-4 "single chain" antibody on allogeneic tumor cells resulted in a reduction of T cell mediated elimination of these tumor cells in mice (Hwang KW, Sweatt WB, Brown IE, Blank C, Gaj ewski TF, Bluestone JA, Alegre ML, Cutting edge: targeted ligation of CTLA-4 in vivo by membrane-bound anti-CTLA-4 antibody to prevent rejection of allogeneic cells. J Immunol, 2002.169 (2): 633-7.) These results indicated that an immunological anti-tumor response or rejection of allogeneic organ transplants through efficient

Cross-linking of CTLA-4 can be suppressed. However, this type of targeted inhibition of T cell activation by CTLA-4 ligation in vivo has so far only been achieved with membrane-bound anti-CTLA-4 antibody constructs or with the natural membrane-bound ligands.

A corresponding suppression of the T cell response in animals by soluble anti-CTLA-4 antibody is not yet described. Only the in vitro induction of apoptosis in preactivated T cells was described by a CTLA-4 antibody specific for the C 'D loop of the extracellular domain of CTLA-4 (Gribben JG, Freeman GJ, Boussiotis VA, Rennert P, Jellis CL Greenfield E, Barber M, Restivo VA Jr, X Ke, Gray GS, Nadler LM, CTLA4 mediates antigen-specific apoptosis of human T cells, Proc Natl Acad Sci USA, 1995. 92 (3): 811-5).

In summary, the findings so far show an inhibitory function of CTLA-4 on T cells, whose mechanism of action is not fully understood. The following mechanisms, which are not mutually exclusive, are discussed: i) suppression of the activating TCR and / or CD28 signaling pathway, ii) competition of CD28-mediated costimulation through higher affinity for CD80 and CD86, iii) increasing the threshold of T cell activation, iv) attenuating T cell expansion, and / or v) activating regulatory T cells and, thus, indirectly inhibiting conventional T cells.

Targeted and well-tolerated inactivation of T cells by stimulating the inhibitory function of CTLA-4 would be desirable in the above-mentioned indications.

Technical problem of the invention.

The invention is therefore based on the technical problem of providing substances and pharmaceutical compositions capable of stimulating the inhibitory function of CTLA-4.

Broad features of the invention and preferred embodiments.

To solve this technical problem, the invention teaches an isolated monoclonal antibody specific and agonistic to CTLA-4, the heavy chain of the antibody containing a sequence selected from the group consisting of (Seq. ID): "22 , 23, 24, 25, 26, 27, 28, 29, 30, and 32 ". The light chain of the antibody may contain a sequence selected from the group consisting of (Seq. ID):" 33, 34 , 35, 36, 37, and 38 ".

An antibody according to the invention with a heavy chain containing a sequence according to Seq. ID 27, 28, or 29, preferably containing or consisting of Sequence according to Seq. ID 30 or 32, and with a light chain containing a sequence according to Seq. ID 36 or 37, preferably containing or consisting of a sequence according to Seq. ID 38.

A specific antibody having the above basic structure are the following antibodies TGN2122. H and TGN2422. H .

Furthermore, the invention teaches an isolated monoclonal antibody which is specific and agonistic to CTLA-4, the heavy chain of the antibody containing a sequence selected from the group consisting of (Seq. ID): "43, 44," 45, 46, 47, 48, 49, 50, 51, and 53 ". The light chain of the antibody may contain a sequence selected from the group consisting of (Seq. ID):" 54, 55, 56, 57, 58, and 59 ^λΛ .

Preference is given to an antibody according to the invention with a heavy chain containing a sequence according to Seq. ID 48, 49, or 50, preferably containing or consisting of the sequence according to Seq. ID 51 or 53, as well as with a light chain containing a sequence according to Seq. ID 57 or 58, preferably containing or consisting of a sequence according to Seq. ID 59.

A specific antibody having the above basic structure are the following antibodies TGN2122. C and TGN2422. C.

The antibodies mentioned above are humanized antibodies. Since it is already one humanized antibody, as described below for other variants of antibodies according to the invention is not required. The antibody may or may not bind to the C '' D loop of CTLA-4. Rather, it may also be an antibody that does not bind to this loop. With regard to all other embodiments as well as uses and other details and explanations, the following attached to a further variant of the invention explanations apply analogously and fully.

Finally, the invention teaches an isolated monoclonal antibody which is specific and agonistic to CTLA-4, wherein the antibody does not bind to a CTLA-4 partial sequence according to SEQ. ID 1 binds. In the sequence according to SEQ. ID 1 is the C 'D loop of the CTLA-4. In other words, an antibody of the invention binds to other regions of the CTLA-4 molecule than the C "D loop. The invention is based on the finding that an agonistic stimulation of CTLA-4, i. e. inducing the inhibitory activity of CTLA-4 in vivo, constitutes a useful therapeutic concept for autoimmune diseases or transplantation, and provides suitable drugs in the form of the antibodies or fragments thereof.

Preferably, an antibody according to the invention contains at least one of the sequences according to SEQ. ID 2 to SEQ. ID 7 or SEQ. ID 8 to SEQ. -ID 13. These sequences are the CDRs of the heavy and light chain variable regions, with additional reference to Table 2. Preferably, an antibody according to the invention is humanized. This can be done in a customary manner, for example by using a human CTLA-4-specific monoclonal mouse antibody is chimaerized in such a way that the constant regions are exchanged for human or human-compatible constant regions. It is essential that preferably all CDRs according to Table 2 are maintained and indeed in their spatial arrangement to each other. As a basis for the humanization, for example, monoclonal antibodies can be used which at least one, preferably all, sequences according to SEQ. ID 2 to SEQ. ID 7 or SEQ. ID 8 to SEQ. ID 13, for example one of the sequences according to SEQ. ID 14 to 17, included. Specifically, antibodies according to the invention can also be one of the sequences according to SEQ. ID 18 to 21 included. Suitable embodiments of antibodies which form a basis for humanization are the antibodies 4.8H10H5 and 4.3F6B5 described in detail below. A preparation of humanized antibodies from this is possible in a conventional manner, for example by genetic engineering humanization strategies.

In the context of the invention, the term "antibody" encompasses, in addition to the structures explicitly disclosed, also functionally corresponding antibodies which have been modified, for example, by chimerization, humanization or de-immunization (excision of T-cell epitopes from the human antibody which trigger undesired immune reactions) and specific antibodies Fragments of the light and / or heavy chain variable region underlying antibody of the above type. The production resp. Obtaining such antibodies with given immunogens is the A person of ordinary skill in the art is familiar and need not be explained in more detail.

The invention also relates to an isolated protein or peptide containing at least one of the sequences SEQ. ID 2 to 13, in particular one of the sequences SEQ. ID 14 to 17 or SEQ. ID 18 to 21, or one of the sequences Seq. ID 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39, in particular one of the sequences Seq. ID 27, 28, 29, 30, 32, 36, 37, or 38, or one of the sequences Seq. ID 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60, in particular one of the sequences Seq. ID 48, 49, 50, 51, 53, 57, 58, or 59, or consisting of one of said sequences, an isolated nucleic acid coding for such a protein or peptide or for a light chain and / or heavy chain of an antibody according to the invention, an isolated vector containing such a nucleic acid, and an isolated cell transfected with such a vector. The above objects are all for the production or. Construction of antibodies according to the invention suitable.

An inventive antibody or. a protein or peptide according to the invention is preferably soluble in water, in particular in physiological saline solution, i. e. not artificially networked. Furthermore, an antibody according to the invention is superagonistic, i. e. stimulates the physiological action of the T-cell inhibitory receptor CTLA-4.

The invention further relates to a pharmaceutical composition containing a monoclonal antibody according to the invention and / or an inventive Protein or peptide, and optionally at least one physiologically acceptable carrier and / or excipient, which will be explained in detail later. It is obtainable by mixing these components, the active ingredient being used in a physiologically effective dose. This dose can be easily determined in in-vitro experiments with cells and in animal experiments in the usual way. Such a pharmaceutical composition is for the prophylaxis and / or therapy of a disease or condition selected from the group consisting of "rheumatoid arthritis, type I diabetes, multiple sclerosis, systemic lupus erythematosus, psoriasis, ulcerative colitis, Crohn's disease, allergies, rejection of allograft organ transplant , in particular organ transplants of the following organs: heart, kidney, liver, pancreas, lung, bone marrow, and "graft-versus-host" disease suitable. Thus, the invention also includes a method for the prophylaxis and / or treatment of a above disease or condition wherein a patient is given the pharmaceutical composition in suitable dosage.

The galenic preparation of a pharmaceutical composition according to the invention can be carried out in the usual way. Suitable counterions for ionic compounds are, for example, Na ⁺ , K ⁺ , Li ⁺ or cyclohexylammonium. Suitable solid or liquid galenic preparations are, for example, granules, powders, dragees, tablets, (micro) capsules, suppositories, syrups, juices, suspensions, emulsions, drops or solutions for injection (i.v., i.p., i .m., s. c.) or nebulization (aerosols), transdermal systems, as well as preparations with protracted release of active ingredient, in their preparation conventional adjuvants such as carriers, blasting, binding, coating, swelling, lubricants or lubricants, flavoring agents, sweeteners and solubilizers, are used. As adjuvants may be magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talcum,

Milk protein, gelatin, starch, cellulose and its derivatives, animal and vegetable oils such as cod liver oil, sunflower, peanut or sesame oil, polyethylene glycols and solvents, such as sterile water and monohydric or polyhydric alcohols, such as glycerol, called.

The invention further relates to a method for producing a monoclonal antibody according to the invention, wherein a nucleic acid according to the invention is introduced into a vector, wherein a cell is transfected by means of the vector, wherein the transfected cell is cultured, wherein a supernatant is separated from the cultured cells or the cultured cell is lysed and the lysate is recovered, and the monoclonal antibodies are separated from the separated supernatant or lysate.

In the following, the invention will be explained in more detail by means of examples which represent only embodiments,

Example 1: Antibodies according to the invention and

Comparison Antibodies

Table 1 shows the binding properties of 4 new anti-CTLA-4 antibodies, 2 of which do not bind to the C "D loop of CTLA-4 (4.3F6B5 and 4.8H10H5) and 2 however, bind to it (3.7F10A2 and 4.7A8H6). The latter are reference antibodies and are not the subject of the present invention. The specificity of the antibodies to human CTLA-4 was investigated, both on transfected Jurkat E6.1 cells and on ex vivo activated human PBMCs (peripheral blood mononuclear cells). Cross-reactivity against rat CTLA-4 was demonstrated with a transfected BW cell line carrying the extracellular domain of rat CTLA-4 on the surface. Likewise, the cross-reactivity against the closely related T-

Cell receptors CD28 and ICOS on transfected Jurkat E6.1 resp. L929 cells excluded. The binding or non-binding to the lateral C ^VN D loop structure is described in detail in FIG. 2 shown. The thick curves represent CTLA-4 and the thin curves represent the isotype control.

Fig. 1 shows the most important examples

Binding characteristics of the inventive anti-CTLA-4 antibody 4.8H10H5 and the reference antibody 3.7F10A2. (A) Transfected Jurkat E6.1 cells carrying a chimeric CTLA-4 / CD28 receptor on their surface were used to identify CTLA-4 specific antibodies. This consists of the extracellular domain of human CTLA-4, which establishes the specificity of the antibodies, and the transmembrane and intracellular domain of mouse CD28. The CD28 part of the receptor ensures stable surface expression of the chimeric receptor. Shown is the binding of antibodies to transfected cells (thick curve) compared to binding to non-transfected cells (thin curve). (B) The specificity of antibodies to CTLA-4 was confirmed with human PBMCs previously stimulated ex vivo with PHA / IL-2. In resting cells, CTLA-4 is predominant localizes intracellularly and is brought to the surface only after activation. The thick curve shows the binding of the antibodies, the thin curve the binding of the isotype control to activated human PBMCs. (C) In view of a possible usability of the antibodies in

Animal models demonstrated cross-reactivity to rat CTLA-4. Transfected BW cells carrying a chimeric human CTLA-4 / mouse CD28 receptor on their surface were used for this purpose. Shown is the binding of the antibody to transfected cells (thick curve) or. to non-transfected cells (thin curve).

Fig. Figure 2 shows the nonexistent specificity of 2 of 4 anti-CTLA-4 antibodies from Table 1 for the C "" D loop. Surprisingly, it was found that the two antibodies according to the invention 4.3F6B5 and 4.8H10H5, which showed agonistic mode of action in functional assays (see Example 2), are not specific for the human C 'D loop. For this purpose Jurkat E6.1 was used, which expresses a chimeric extracellular domain of CTLA-4 on the surface: this chimera consists of the murine receptor, in which the C ^S D loop against the corresponding human sequence, represented by the amino acids Pos 68 - 83, was exchanged. The binding of the antibodies 3.7F10A2 and 4.7A8H6 to the C 'D loop shows that the construct was efficiently expressed. For the antibodies 3.7F10A2 and 4.7A8H6, this amino acid sequence is sufficient for binding (thick line, thin curve: isotype control). For antibodies 4.8H10H5 and 4.3F6B5 this sequence is not sufficient for binding.

Fig. Figure 3 shows that binding of anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 to CTLA-4 by addition of recombinant CD80 can be competed. The Ergbenis suggests that 4.8H10H5 and 4.3F6B5 are located near the binding site for CD80, d. H . bind the MYPPPY loop, but not to the C 'D loop. The aim of this experiment was to further narrow the binding properties of the antibodies. Jurkat E6.1 cells carrying the extracellular domain on their surface were incubated with increasing concentration of CD80Fc protein and 1 μg / ml of CTLA-4 specific antibody. The co-incubation of recombinant protein and antibody leads to a

Displacement of the binding of antibodies 4.8H10H5 and 4.3F6B5 to the extracellular domain of CTLA-4.

Experiments as described for FIG. 1 were done accordingly for the antibodies TGN2122. C, TGN2422. C.,

TGN2122. H and TGN2422. H performed. Figure 10 shows the binding of the humanized antibodies to the extracellular domain of human CTLA-4. Shown is the binding of the humanized antibodies to transfected cells (thick curve) compared to binding of the isotype control (thin curve) to the same cells. The FACS analysis shows that in the course of humanization the specificity of the antibodies to human CTLA-4 is maintained.

Table 2 gives the sequences of the invention

Antibodies 4.8H10H5 and 4.3F6B5 again, divided into heavy and light chains, with the border between the variable regions and the constant regions labeled. The sequences were determined by RT-PCR or. determined by protein sequencing (Edman degradation).

Table 3 shows heavy chain sequences of the antibody TGN2122. H . Table 4 shows the for the heavy chain coding nucleic acid. Table 5 shows heavy chain sequences of the antibody TGN2422. H . Table 6 shows the heavy chain coding nucleic acid. Table 7 shows light chain sequences for both antibodies TGN2122. H and TGN2422. H . Table 8 shows the light chain encoding nucleic acid.

Table 9 shows heavy chain sequences of the antibody TGN2122. C. Table 10 shows the heavy chain coding nucleic acid. Table 11 shows heavy chain sequences of the antibody TGN2422. C. Table 12 shows the heavy chain coding nucleic acid. Table 13 shows light chain sequences for both antibodies TGN2122. C and TGN2422. C. Table 14 shows the light chain encoding nucleic acid.

In the sequences Seq. IDs 31, 39, 52, and 60 are leader peptides that are not included in the current mature chains. Therefore, those antibodies are preferred which do not contain these sequences.

The antibodies TGN2122. C (isotype IgG1) and TGN2422. C (isotype IgG4) were purified from mouse antibody 4.3F6B5

Humanization received. The antibodies TGN2122. H (isotype IgG1) and TGN2422. H (isotype IgG4) were obtained from mouse antibody 4.8H10H5.

Example 2: Effect of Antibodies According to the Invention on a Comparative Antibody Attached to C 'D Loop binds, as well as commercially available anti-CTLA-4 antibodies.

Fig. Figure 4 shows the inhibitory effect of anti-CTLA-4 antibody 4.8H10H5 on the proliferation of human PBMCs.

The aim of this proliferation inhibition assay was to identify a functionally novel antibody compared to previously known CTLA-4 specific antibodies. As an essential property of a superagonistic antibody has been defined the ability to reduce the proliferation of human PBMC. Another criterion was that this effect with soluble, not artificially networked! Antibody is observed. These antibodies were evaluated as positive, reducing anti-CD3 (or superagonistic anti-CD28, not shown) T cell proliferation by at least 25%. Readout system was the measurement of proliferation by means of ³ H thymidine incorporation. In this assay system, the CTLA-4 specific antibodies were given in time with the activating anti-CD3 antibody and the proliferation determined after 63-66 hours. Antibody 4.8H10H5 was able to inhibit proliferation of T cells based on the above criteria. For comparison, an antibody not positively evaluated in this assay system (2.10B11A1) is listed. Shown is the relative proliferation in relation to the positive control (anti-CD3 induced proliferation). Further controls included the respective isotype control (IgGl or IgG2) and a commercially available antibody (BNI3, BD Pharmingen). The control commercial antibodies (14D3, 8H5, 3H1833, BNI3) with specificity for CTLA-4 had no effect. Fig. Figure 5 shows the stimulatory effect of the anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 on the IL-2 production of Jurkat E6.1 cells expressing a chimeric CTLA-4 / CD28 molecule. Jurkat E6.1 cells expressing a chimeric CTLA-4 / CD28 receptor on their surface were used for this cell-autonomous read-out system for the functional characterization of the CTLA-4-specific MAK (D). This consists of the extracellular domain of the CTLA-4 receptor and the transmembrane and intracellular domain of CD28. Activation of the chimeric receptor by CTLA-4-specific MAK induces CD28-specific activation markers such as IL-2 or CD69, which are detected by means of ELISA or ELISA. FACS analysis can be measured. Potentially superagonistic CTLA-4 specific antibodies can be identified in this system using CD28 specific

Activation markers are identified. Control antibodies and CTLA-4 specific antibodies were cross-linked (using sheep anti mouse Ig) and incubated with 1 * 10 ⁵ transfected Jurkat E6.1 cells for 48h. As an activation marker, IL-2 production was measured by ELISA.

Isotype controls and commercially available antibodies with specificity for CTLA-4 were included as controls. (A) Effect of CTLA-4 specific antibodies (1 μg / ml) on the IL-2 production of transfected Jurkat cells expressing a chimeric CTLA-4 / CD28 receptor. (B) Effect of commercially available CTLA-4 specific antibodies (1 μg / ml) on the IL-2 production of transfected Jurkat cells expressing a chimeric CTLA-4 / CD28 receptor. (C) Effect of CTLA-4 specific antibodies (lμg / ml) on IL-2 production of untransfected Jurkat E6.1 cells lacking the chimeric receptor (representative experiments). Two of the antibodies tested (4.3F6B5, 4.8H10H5) induce the IL-2 production of transfected Jurkat cells by activation of the chimeric receptor, while none of the commercially available antibodies with specificity for CTLA-4 was able to do so.

Fig. Figure 6 shows the stimulatory effect of the anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 on the CD69 induction of Jurkat E6.1 cells expressing a chimeric CTLA-4 / CD28 molecule. (D) Using the method shown in Fig. In addition to IL-2 production, the assay of CD69 expression as a further activation marker was measured by FACS analysis. In contrast to IL-2 production, CD69 is an early activation marker and can be detected after 4 h of incubation of the antibodies with the transfected cells. (A) Effect of CTLA-4-specific antibodies (1 μg / ml) on transfected CD69 expression Jurkat cells expressing a chimeric CTLA-4 / CD28 receptor (B) Effect of commercially available CTLA-4 specific antibodies (1 μg / ml) on the CD69 expression of transfected Jurkat cells carrying a chimeric CTLA-4 / CD28 receptor (C) Effect of CTLA-4 specific antibodies (lμg / ml) on CD69 expression of untransfected Jurkat Eβ.l cells lacking the chimeric receptor (representative experiments) As shown for IL-2, as well Antibodies 4.3F6B5 and 4.8H10H5 are able to activate the chimeric receptor, a

Trigger signal transduction and induce CD69 expression. None of the commercially available antibodies with specificity for CTLA-4 was able to do this (representative experiments).

In Fig. Figure 7 shows that the stimulatory effect of the anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 on the chimeric CTLA-4 / CD28 construct by CD80 Fc protein can be reduced. Jurkat E. β. 1 cells expressing the chimeric CTLA-4 / CD28 receptor (see Fig. 5, 6) were incubated with increasing concentration of antibody and in each case 1 μg / ml CD80 Fc protein. Antibody-mediated CD69 expression is then reduced by CD80Fc as the recombinant protein is excessively incubated compared to the antibody. Rectangular squares show the current CD69 induction without CD80 co-incubation, the triangular curves show the CD69 induction diminished by Iμg / ml CD80 Fc

Antibodies. In addition, the concentration-dependent binding of the antibodies and the Jurkat cells is shown.

In Fig. Figure 8 shows the cross-reactivity of the anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 with the rat CTLA-4 molecule and the stimulatory functionality of the antibodies in a chimeric receptor assay. (A) Binding of anti-CTLA-4 antibodies to rat CTLA-4 was demonstrated by BW cells expressing the extracellular domain of the rat receptor on its surface. Analogous to the chimeric receptor on Jurkat cells (Figure 5), these cells express a chimeric CTLA-4 CD28 receptor consisting of rat CTLA-4 (extracellular domain) and mouse CD28 (transmembrane / intracellular domain) (bright line: anti-CTLA). 4 Akö, dark curve: isotype control). (B) For the cross-reactive antibodies 4.3F6B5 and 4.8H10H5, activation of the chimeric receptor was demonstrated by IL-2 induction. As in FIG. 5 can be identified with the transfected BW cells potentially superagonistic CTLA-4 specific antibodies by means of CD28 specific activation markers (IL-2). In a control experiment the antibodies on BW cells lacking the chimeric receptor could not induce IL-2 production.

Fig. Figure 9 shows the inhibitory in vivo effect of anti-CTLA-4 antibodies 4.8H10H5 and 4.3F6B5 on CD28

SuperMAB-mediated activation of T cells in the rat. For this purpose, activating superagonistic rats of specific CD28 antibodies (JJ316) together with CTLA-4 specific antibodies resp. Isotype Control Rats i. v. applied. After three days, cell suspensions were eliminated

The lymph nodes and spleens were obtained and analyzed by FACS for the activation marker CD25. A total of three experiments with varying antibody concentrations were performed. Both tested CTLA-4 specific antibodies reduced the JJ316-induced CD25 expression on lymph node as well as on spleen cells in 3 independent experiments by approx. 30 - 40%. Shown is a representative result.

In Figure 11, the stimulatory effect of the humanized anti-CTLA-4 antibody (1 μg / ml) in vitro on the CD69 expression of Jurkat E6.1 cells expressing a chimeric CTLA-4 / CD28 receptor is shown. The procedure was as described for FIG. 6. From Figure 11 it can be seen that all humanized

Antibodies of the invention, from both IgG1 and IgG4 isotypes, effectively induce CD69 surface expression while the isotype control resp. the addition of cell culture medium without effect remained (representative result). This shows that the functionally novel properties of the antibodies 4.3F6B5 and 4.8H10H5 have been preserved in the course of humanization. FIG. 12 shows the inhibitory effect of the antibody TGN2122. C and TGN2122. H on the proliferation of ex vivo stimulated human PBMCs. In a recall-response-assay 10 ^Λ 5 human PBMCs of healthy donors with 2, 5 μg / ml tetanus toxoid were activated and at the same time the corresponding CTLA-4 specific antibodies resp. the isotype control given to the experimental approach. The proliferation was measured by ³ H thymidine incorporation after 120 h incubation. For this purpose, ³ H-thymidine was added to the experimental batch for the last 15-18 h of the test duration and the ³ H-Thymidineinbau determined. As controls, the corresponding isotype control and an approach in which unactivated non-antibody cells were included were included. Both antibodies were able to effectively inhibit tetanus toxoid-induced proliferation. In contrast, the isotype control showed no such inhibition. With these experiments, the superagonistic properties of antibodies according to the invention are shown since inhibition in soluble form, i. e. without artificial cross-linking, took place.

If _{n is} specified in sequences X, the specified n can vary by + 1.

Table 1 Binding characteristics of new anti-CTLA-4 antibodies

Tab. 2

4.3F6B5 Heavy Chain

CDR-HL CDR-H2

EVQLQPSGPVL VKPGÄSVKISCKASPYTFTDYKIPJWVKQSHGKSLEWIGtf IYPYSGSSDYNOK

FKsJKÄTLTVDNSSTTAYMELRSLTSEDSAVYYCAR ^ GbAMDYiiΛi'GQGTAVTVSS

CDR-H3

Light chain

CDR-L1 CDR-L2

DILMTQSPASLSASVGETVTITClSASENIYGALΪWYQRKQGKSPQLLIYiSATNLAElGMSSRFS

4.8H10H5 Heavy chain

CDR-HL CDR-H2

DVKL VESGGGL VKLGGSLKLSCAASfSFT FNI YYMS} WVRQTPEKRLELV £ | AINPDGGNTYYPDT

VCG ^ FTISRDNAKNTLYLQMSSLKSEDSALYYCARlϊGGPGFDsiΛfGQGTTLTVSS

CDR-H3

Light chain

CDR-L1 CDR-L2

ENVLTQSPAIMSASPGERVTMTCßASSSVSYMHlWYQQKSNTSPKLWIYiDTSKLAsiGVPGRFSG

SGSRNS YSLTISSMEAEDVATYYCIFPGSGFPFMYTJFGGGTKLEIKR

CDR-L3 TAB 3

TGN2122. H HC amino acid sequence

Seq. 30 EVQLVE_SGGGL_V_QP ^

GLELVAAINPpGGNTYYPDTVKGRFT SWGQGTLVTVSSASTKGPSVFP] -APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV

HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPP SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Seq. 31:

MGWSCIILFLVATATGVHS = Leader peptide, not part of mature HC

XXXX = variable segment XXXX = constant segment

CDRs

Kabat IYYMS - Seq. 22

AbM GFTFNYYMS - Seq. 25

EVQLVESGGGLVQPGGSLRLSCAASGFTFNIYYMSWVRQAPGKGLELVA

AINPDGGNTYYPDTVKG - Seq. 23

AINPDGGNTY - Seq. 26

AINPDGGNTYYPDTVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR

YGGPGFDS - Seq. 24 YGGPGFDS - Seq. 24 YGGPGFDSWGQGTLVTVSS - Seq. 27

IYYMSX ₁₄ AINPDGGNTYYPDTVKGX ₃₂ YGGPGFDS - Seq. 28 GFTFNIYYMSX ₁₄ AINPDGGNTYX ₃₉ YGGPGFDS - Seq. 29 TAB 4

TGN2122. H HC nucleotide sequence (SEQ ID NO: 40)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA

51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGACAAT

101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCG AGGTGCAGCT

151 GGTGGAGAGC GGCGGCGGCC TGGTGCAGCC CGGCGGCAGC CTGAGGCTGA

201 GCTGCGCCGC CAGCGGCTTC ACCTTCAACA TCTACTACAT GAGCTGGGTG

251 AGGCAGGCCC CCGGCAAGGG CCTGGAGCTG GTGGCCGCCA TCAACCCCGA

301 CGGCGGCAAC ACCTACTACC CCGACACCGT GAAGGGCAGG TTCACCATCA

351 GCAGGGACAA CGCCAAGAAC AGCCTGTACC TGCAGATGAA CAGCCTGAGG

401 GCCGAGGACA CCGCCGTGTA CTACTGCGCC AGGTACGGCG GCCCCGGCTT

451 CGACAGCTGG GGCCAGGGCA CCCTGGTGAC CGTGAGCAGC GGTGAGTCGT

501 ACGCTAGCAA GCTTTCTGGG GCAGGCCAGG CCTGACCTTG GCTTTGGGGC

551 AGGGAGGGGG CTAAGGTGAG GCAGGTGGCG CCAGCCAGGT GCACACCCAA

601 TGCCCATGAG CCCAGACACT GGACGCTGAA CCTCGCGGAC AGTTAAGAAC

651 CCAGGGGCCT CTGCGCCCTG GGCCCAGCTC TGTCCCACAC CGCGGTCACA

701 TGGCACCACC TCTCTTGCAG CCTCCACCAA GGGCCCATCG GTCTTCCCCC

751 TGGCACCCTC CTCCAAGAGC ACCTCTGGGG GCACAGCGGC CCTGGGCTGC

801 CTGGTCAAGG ACTACTTCCC CGAACCGGTG ACGGTGTCGT GGAACTCAGG

851 CGCCCTGACC AGCGGCGTGC ACACCTTCCC GGCTGTCCTA CAGTCCTCAG

901 GACTCTACTC CCTCAGCAGC GTGGTGACCG TGCCCTCCAG CAGCTTGGGC

951 ACCCAGACCT ACATCTGCAA CGTGAATCAC AAGCCCAGCA ACACCAAGGT

1001 GGACAAGAAA GTTGGTGAGA GGCCAGCACA GGGAGGGAGG GTGTCTGCTG

1051 GAAGCCAGGC TCAGCGCTCC TGCCTGGACG CATCCCGGCT ATGCAGCCCC

1101 AGTCCAGGGC AGCAAGGCAG GCCCCGTCTG CCTCTTCACC CGGAGGCCTC

1151 TGCCCGCCCC ACTCATGCTC AGGGAGAGGG TCTTCTGGCT TTTTCCCAGG

1201 CTCTGGGCAG GCACAGGCTA GGTGCCCCTA ACCCAGGCCC TGCACACAAA

1251 GGGGCAGGTG CTGGGCTCAG ACCTGCCAAG AGCCATATCC GGGAGGACCC

1301 TGCCCCTGAC CTAAGCCCAC CCCAAAGGCC AAACTCTCCA CTCCCTCAGC

1351 TCGGACACCT TCTCTCCTCC CAGATTCCAG TAACTCCCAA TCTTCTCTCT

1401 GCAGAGCCCA AATCTTGTGA CAAAACTCAC ACATGCCCAC CGTGCCCAGG

1451 TAAGCCAGCC CAGGCCTCGC CCTCCAGCTC AAGGCGGGAC AGGTGCCCTA

1501 GAGTAGCCTG CATCCAGGGA CAGGCCCCAG CCGGGTGCTG ACACGTCCAC

1551 CTCCATCTCT TCCTCAGCAC CTGAACTCCT GGGGGGACCG TCAGTCTTCC

1601 TCTTCCCCCC AAAACCCAAG GACACCCTCA TGATCTCCCG GACCCCTGAG

1651 GTCACATGCG TGGTGGTGGA CGTGAGCCAC GAAGACCCTG AGGTCAAGTT

1701 CAACTGGTAC GTGGACGGCG TGGAGGTGCA TAATGCCAAG ACAAAGCCGC

1751 GGGAGGAGCA GTACAACAGC ACGTACCGGG TGGTCAGCGT CCTCACCGTC

1801 CTGCACCAGG ACTGGCTGAA TGGCAAGGAG TACAAGTGCA AGGTCTCCAA

1851 CAAAGCCCTC CCAGCCCCCA TCGAGAAAAC CATCTCCAAA GCCΆAAGGTG

1901 GGACCCGTGG GGTGCGAGGG CCACATGGAC AGAGGCCGGC TCGGCCCACC

1951 CTCTGcccTG AGAGTGACCG CTGTACCAΆC CTCTGTCCCT ACAGGGCAGC

2001 CCCGAGAACC ACAGGTGTAC ACCCTGCCCC CATCCCGGGA TGAGCTGACC 2051 AAGAACCAGG TCAGCCTGAC CTGCCTGGTC AAAGGCTTCT ATCCCAGCGA 2101 CATCGCCGTG GAGTGGGAGA GCAATGGGCA GCCGGAGAAC AACTÄCAAGA 2151 CCACGCCTCC CGTGCTGGAC TCCGACGGCT CCTTCTTCCT CTACAGCAAG 2201 CTCACCGTGG ACAAGAGCAG GTGGCAGCAG GGGAACGTCT TCTCATGCTC 2251 CGTGATGCAT GAGGCTCTGC ACAACCACTA CACGCAGAAG AGCCTCTCCC 2301 TGTCTCCGGG TAAATGA . ^ ^U - ßies

TGN2422. H HC amino acid sequence

Seq. 32: EV _Q LVESGGGLV _Q ^

GLEL V ^ INPDGGNTYYPDTVKG3_FTI SRDNAKN

SWGGGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGV

HTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAP

EFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE

QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQE

EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ

EGNVFSCSVMHEALHNHYTQKSLSLSLGK

Seq. 31:

MGWSCIILFLVATÄTGVHS = Leader peptide, not part of the mature HC

XXXX = variable segment XXXX = constant segment

CDRs

Kabat IYYMS - Seq. 22

AbM GFTFNYYMS - Seq. 25

EVQLVESGGGLVQPGGSLRLSCAASGFTFNIYYMSWVRQAPGKGLELVA

AINPDGGNTYYPDTVKG - Seq. 23

AINPDGGNTY - Seq. 26

AINPDGGNTYYPDTVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR

YGGPGFDS - Seq. 24 YGGPGFDS - Seq. 24 YGGPGFDSWGQGTLVTVSS - Seq. 27 TAB 6

TGN2422. H HC nucleotide sequence (SEQ ID NO: 41)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA 51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGACAAT 101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCG AGGTGCAGCT 151 GGTGGAGAGC GGCGGCGGCC TGGTGCAGCC CGGCGGCAGC CTGAGGCTGA 201 GCTGCGCCGC CAGCGGCTTC ACCTTCAACA TCTACTACAT GAGCTGGGTG 251 AGGCAGGCCC CCGGCAAGGG CCTGGAGCTG GTGGCCGCCA TCAACCCCGA 301 CGGCGGCAAC ACCTACTACC CCGACACCGT GAAGGGCAGG TTCACCATCA 351 GCAGGGACAA CGCCAAGAAC AGCCTGTACC TGCAGATGAA CAGCCTGAGG 401 GCCGAGGACA CCGCCGTGTA CTACTGCGCC AGGTACGGCG GCCCCGGCTT 451 GGCCAGGGCA CCCTGGTGAC CGACAGCTGG CGTGAGCAGC GGTGAGTCGT 501 ACGCTAGCAA GCTTTCTGGG GCAGGCCGGG CCTGACTTTG GCTGGGGGCA 551 GGGAGGGGGC TAAGGTGACG CAGGTGGCGC CAGCCAGGTG CACACCCAAT 601 GCCCATGAGC CCAGACACTG GACCCTGCAT GGACCATCGC GGATAGACAA 651 GAACCGAGGG GCCTCTGCGC CCTGGGCCCA GCTCTGTCCC ACACCGCGGT

701 CACATGGCAC CACCTCTCTT GCAGCTTCCA CCAAGGGCCC ATCCGTCTTC 751 CCCCTGGCGC CCTGCTCCAG GAGCACCTCC GAGAGCACAG CCGCCCTGGG

801 CTGCCTGGTC AAGGACTACT TCCCCGAACC GGTGACGGTG TCGTGGAACT 851 CAGGCGCCCT GACCAGCGGC GTGCACACCT TCCCGGCTGT CCTACAGTCC 901 TCAGGACTCT ACTCCCTCAG CAGCGTGGTG ACCGTGCCCT CCAGCAGCTT 951 GGGCACGAAG ACCTACACCT GCAACGTAGA TCACAAGCCC AGCAACACCA

1001 AGGTGGACAA GAGAGTTGGT GAGAGGCCAG CACAGGGAGG GAGGGTGTCT 1051 GCTGGAAGCC AGGCTCÄGCC CTCCTGCCTG GACGCACCCC GGCTGTGCAG 1101 CCCCAGCCCA GGGCAGCAAG GCATGCCCCA TCTGTCTCCT CACCCGGAGG 1151 CCTCTGACCA CCCCACTCAT GCTCAGGGAG AGGGTCTTCT GGATTTTTCC 1201 ACCAGGCTCC GGGCAGCCAC AGGCTGGATG CCCCTACCCC AGGCCCTGCG 1251 CATACAGGGG CAGGTGCTGC GCTCAGACCT GCCAAGAGCC ATATCCGGGA 1301 GGACCCTGCC CCTGACCTAA GCCCACCCCA AAGGCCAAAC TCTCCACTCC 1351 CTCAGCTCAG ACACCTTCTC TCCTCCCAGA TCTGAGTAAC TCCCAATCTT 1401 CTCTCTGCAG AGTCCAAATA TGGTCCCCCA TGCCCATCAT GCCCAGGTAA 1451 GCCAACCCAG GCCTCGCCCT CCAGCTCAAG GCGGGACAGG TGCCCTAGAG 1501 TAGCCTGCAT CCAGGGACAG GCCCCAGCCG GGTGCTGACG CATCCACCTC 1551 CATCTCTTCC TCAGCACCTG AGTTCCTGGG GGGACCATCA GTCTTCCTGT 1601 TCCCCCCAAA ACCCAAGGAC ACTCTCATGA TCTCCCGGAC CCCTGAGGTC 1651 ACGTGCGTGG TGGTGGACGT GAGCCAGGAA GACCCCGAGG TCCAGTTCAA 1701 CTGGTACGTG GATGGCGTGG AGGTGCATAA TGCCAAGACA AAGCCGCGGG 1751 AGGAGCAGTT CAACAGCACG TACCGTGTGG TCAGCGTCCT CACCGTCCTG 1801 CACCAGGACT GGCTGAACGG CAAGGAGTAC A AGTGCAAGG TCTCCAACAA

1851 AGGCCTCCCG TCCTCCATCG AGAΆAACCAT CTCCAAAGCC AΆAGGTGGGA

1901 CCCACGGGGT GCGAGGGCCA CATGGACAGA GGTCAGCTCG GCCCACCCTC

1951 TGCCCTGGGA GTGACCGCTG TGCCAACCTC TGTCCCTACA GGGCAGCCCC

2001 GAGAGCCACA GGTGTACACC CTGCCCCCAT CCCAGGAGGA GATGACCAAG

2051 AACCAGGTCA GCCTGACCTG CCTGGTCAAA GGCTTCTACC CCAGCGACAT

2101 CGCCGTGGAG TGGGAGAGCA ATGGGCAGCC GGAGAACAAC TACAAGACCA

2151 CGCCTCCCGT GCTGGACTCC GACGGCTCCT TCTTCCTCTA CAGCAGGCTA

2201 ACCGTGGACA AGAGCAGGTG GCAGGAGGGG AATGTCTTCT CATGCTCCGT

2251 GATGCATGAG GCTCTGCACA ACCACTACAC ACAGAAGAGC CTCTCCCTGT 2301 CTCTGGGTAA ATGA TAB 7

TGN2122 / TGN2422. H-kappa LC amino acid sequence

SEQ. 38: ENVLT_Q_SPAT_LS_LSPGERAT_LS_CSASS

RLWIYDTSKLASGIPARFSGSGSRNPXTLT

EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ

DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Seq. 39:

MGWSCIILFLVATÄTGVHS = Leader peptide, not part of the mature LC

XXXX = variable segment XXXX = constant segment

CDRs

Here: Kabat = AbM

SASSSVSYMH - Seq. 33 DTS CLASS - Seq. 34 ENVLTQSPATLSLSPGERATLSCSASSSVSYMHWYQQKPGQAPRLWIYDTSKLASGIPARFS

FPGSGFPFMYT - Seq. 35 GSGSRNDYTLTISSLEPEDFAVYYCFPGSGFPFMYTFGGGTKVEIK - Seq. 37

SASSSVSYMHXi ₅ DTSKLASX ₃₂ FPGSGFPFMYT - Seq. 36

³² , 1 u. U, \

TAB 8

TGN2122 / TGN2422. H-kappa LC nucleotide sequence (SEQ 42)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA 51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGACAAT 101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCG AGAACGTGCT 151 GACCCAGAGC CCCGCCACCC TGAGCCTGAG CCCCGGCGAG AGGGCCACCC 201 TGAGCTGCAG CGCCAGCAGC AGCGTGAGCT ACATGCACTG GTACCAGCAG 251 AAGCCCGGCC AGGCCCCCAG GCTGTGGATC TACGACACCA GCAAGCTGGC 301 CAGCGGCATC CCCGCCAGGT TCAGCGGCAG CGGCAGCAGG AACGACTACA 351 CCCTGACCAT CAGCAGCCTG GAGCCCGAGG ACTTCGCCGT GTACTACTGC 401 TTCCCCGGCA GCGGCTTCCC CTTCATGTAC ACCTTCGGCG GCGGCACCAA 451 GGTGGAGATC AAGCGTGAGT CGTACGCTAG CAAGCTTGAT ATCGAATTCT

501 AAACTCTGAG GGGGTCGGAT GACGTGGCCA TTCTTTGCCT AΆAGCΆTTGA

551 GTTTACTGCA AGGTCAGAAA AGCATGCAAA GCCCTCAGAA TGGCTGCAAA 601 GAGCTCCAAC AAAACAATTT AGAACTTTAT TAAGGAATAG GGGGAAGCTA 651 GGAAGAAACT CAAAACATCA AGATTTTAAA TACGCTTCTT GGTCTCCTTG

701 CTATAATTAT CTGGGATAAG CATGCTGTTT TCTGTCTGTC CCTAACATGC 751 CCTGTGATTA TCCGCAAACA ACACACCCAA GGGCAGAACT TTGTTACTTA

801 AACACCATCC TGTTTGCTTC TTTCCTCAGG AACTGTGGCT GCACCATCTG

851 TCTTCATCTT CCCGCCÄTCT GATGAGCAGT TGΆAATCTGG AACTGCCTCT

901 GTTGTGTGCC TGCTGAATAA CTTCTATCCC AGAGAGGCCA AAGTACAGTG

951 GAAGGTGGAT AACGCCCTCC AATCGGGTAA CTCCCAGGAG AGTGTCACAG

1001 AGCAGGACAG CAAGGACAGC ACCTACAGCC TCAGCAGCAC CCTGACGCTG

1051 AGCAAAGCAG ACTACGAGAA ΆCACAAÄGTC TACGCCTGCG AAGTCACCCA

1101 TCAGGGCCTG AGCTCGCCCG TCACAAAGAG CTTCAACAGG GGAGAGTGTT 1151 AG

TAB 9

TGN2122. C HC amino acid sequence

GLEWIGYIYPYSGSSDTO

WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH

TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP

APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR

EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS

RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR

WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Seq. 52

MGWSCIILFLVATATGVHS = Leader peptide, not part of mature HC

XXXX = variable segment XXXX = constant segment

CDRs

Kabat DYKIH - Seq. 43

ABM GYTFTDYKIH - Seq. 46

QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYKIHWVRQAPGQGLEWIG

YIYPYSGSSDYNQKFKS -Seq. 44

YIYPYSGSSD - Seq. 47

YIYPYSGSSDYNQKFKSRATLTVDNSISTAYMELSRLRSDDTAVYYCAR

GGDAMDY - Seq. 45 GGDAMDY - Seq. 45 GGDAMDYWGQGTLVTVSS - Seq. 48

DYKIHX14YIYPYSGSSDYNQKFKSX32GGDAMDY - Seq. 49 GYTFTDYKIHX ₁₄ YIYPYSGSSDX ₃₉ GGDAMDY - Seq. 50 TAB 10

TGN2122. C HC nucleotide sequence (SEQ ID NO: 61)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA 51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGÄCAAT 101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCC AGGTGCAGCT 151 GGTGCAGAGC GGCGCCGAGG TGAAGAAGCC CGGCGCCAGC GTGAAGGTGA 201 GCTGCAAGGC CAGCGGCTAC ACCTTCACCG ACTACAAGAT CCACTGGGTG 251 AGGCAGGCCC CCGGCCAGGG CCTGGAGTGG ATCGGCTACA TCTACCCCTA 301 CAGCGGCAGC AGCGACTACA ACCAGAAGTT CAAGAGCAGG GCCACCCTGA 351 CCGTGGACAA CAGCATCAGC ACCGCCTACA TGGAGCTGAG CAGGCTGAGG 401 AGCGACGACA CCGCCGTGTA CTACTGCGCC AGGGGCGGCG ACGCCATGGA 451 CTACTGGGGC CAGGGCACCC TGGTGACCGT GAGCAGCGGT GAGTCGTACG 501 CTAGCAAGCT TTCTGGGGCA GGCCAGGCCT GACCTTGGCT TTGGGGCAGG 551 GAGGGGGCTA AGGTGAGGCA GGTGGCGCCA GCCAGGTGCA CACCCAATGC

601 ccATGAGccc AGACACTGGA CGCTGAACCT CGCGGACAGT TAΆGAACCCA

651 GGGGCCTCTG CGCCCTGGGC CCAGCTCTGT CCCACACCGC GGTCACATGG

701 CACCACCTCT CTTGCAGCCT CCACCAAGGG CCCATCGGTC TTCCCCCTGG

751 CACCCTCCTC CAAGAGCACC TCTGGGGGCA CAGCGGCCCT GGGCTGCCTG

801 GTCAAGGACT ACTTCCCCGA ACCGGTGÄCG GTGTCGTGGA ACTCAGGCGC

851 CCTGACCAGC GGCGTGCACA CCTTCCCGGC TGTCCTACAG TCCTCAGGAC

901 TCTACTCCCT CAGCAGCGTG GTGACCGTGC CCTCCAGCAG CTTGGGCACC

951 CAGACCTACA TCTGCAACGT GAATCACAAG CCCAGCAACA CCAAGGTGGA

1001 CAAGAAAGTT GGTGAGAGGC CAGCACAGGG AGGGAGGGTG TCTGCTGGAA

1051 GCCAGGCTCA GCGCTCCTGC CTGGACGCAT CCCGGCTATG CAGCCCCAGT

1101 CCAGGGCAGC AAGGCAGGCC CCGTCTGCCT CTTCACCCGG AGGCCTCTGC

1151 CCGCCCCACT CATGCTCAGG GAGAGGGTCT TCTGGCTTTT TCCCAGGCTC

1201 TGGGCAGGCA CAGGCTAGGT GCCCCTAACC CAGGCCCTGC ACACAAAGGG

1251 GCAGGTGCTG GGCTCAGACC TGCCAAGAGC CATATCCGGG AGGACCCTGC

1301 CCCTGACCTA AGCCCACCCC AAAGGCCAAA CTCTCCACTC CCTCAGCTCG

1351 GACACCTTCT CTCCTCCCAG ATTCCAGTAA CTCCCAATCT TCTCTCTGCA

1401 GAGCCCAAAT CTTGTGACAA AACTCACACA TGCCCACCGT GCCCAGGTAA

1451 GCCAGCCCAG GCCTCGCCCT CCAGCTCAAG GCGGGACAGG TGCCCTAGAG

1501 TAGCCTGCAT CCAGGGACAG GCCCCAGCCG GGTGCTGACA CGTCCACCTC

1551 CATCTCTTCC TCAGCACCTG AACTCCTGGG GGGACCGTCA GTCTTCCTCT

1601 TCCCCCCAAA ACCCAAGGAC ACCCTCATGA TCTCCCGGAC CCCTGAGGTC

1651 ACATGCGTGG TGGTGGACGT GAGCCACGAA GACCCTGAGG TCAAGTTCAA

1701 CTGGTACGTG GACGGCGTGG AGGTGCATAA TGCCAAGACA AAGCCGCGGG

1751 AGGAGCÄGTA CAACAGCACG TACCGGGTGG TCAGCGTCCT CACCGTCCTG

1801 CACCAGGACT GGCTGAATGG CAAGGAGTAC AAGTGCAAGG TCTCCAACAA

1851 AGcccTCccA GCCCCCATCG AGAAAACCAT CTCCAAAGCC AAAGGTGGGΆ

1901 CCCGTGGGGT GCGAGGGCCA CATGGACAGA GGCCGGCTCG GCCCACCCTC

1951 TGCCCTGAGA GTGACCGCTG TACCAACCTC TGTCCCTACA GGGCAGCCCC

2001 GAGAACCACA GGTGTACACC CTGCCCCCAT CCCGGGATGA GCTGACCAAG

2051 AACCAGGTCA GCCTGACCTG CCTGGTCAAA GGCTTCTATC CCAGCGACAT

2101 CGCCGTGGAG TGGGAGAGCA ATGGGCAGCC GGAGAACAAC TACAAGACCA

2151 CGCCTCCCGT GCTGGACTCC GACGGCTCCT TCTTCCTCTA CAGCAAGCTC

2201 ACCGTGGACA AGAGCAGGTG GCAGCAGGGG AACGTCTTCT CATGCTCCGT

2251 GATGCATGAG GCTCTGCACA ACCACTACAC GCAGAAGAGC CTCTCCCTGT 2301 CTCCGGGTAA ATGA TAB 11 ^!

TGN2422. C HC amino acid sequence

Seq. 53: QYQkYQSGJ ^ VKKPGA ^

WGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH

TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPE

FLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQ

FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE

MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE

GNVFSCSVMHEALHNHYTQKSLSLSLGK

Seq. 52:

MGWSCIILFLVATATGVHS = Leader peptide, not part of mature HC

XXXX = variable segment XXXX = constant segment

CDRs

Kabat DYKIH - Seq. 43

ABM GYTFTDYKIH - Seq. 46

QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYKIHWVRQAPGQGLEWIG

YIYPYSGSSDYNQKFKS - Seq. 44

YIYPYSGSSD - Seq. 47

YIYPYSGSSDYNQKFKSRATLTVDNSISTAYMELSRLRSDDTAVYYCAR

GGDAMDY - Seq. 45 GGDAMDY - Seq. 45 GGDAMDYWGQGTLVTVSS - Seq. 48 TAB 12

TGN2422. C HC nucleotide sequence (SEQ ID NO: 62)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA 51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGACAAT 101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCC AGGTGCAGCT 151 GGTGCAGAGC GGCGCCGAGG TGAAGAAGCC CGGCGCCAGC GTGAAGGTGA 201 GCTGCAAGGC CAGCGGCTAC ACCTTCACCG ACTACAAGAT CCACTGGGTG 251 AGGCAGGCCC CCGGCCAGGG CCTGGAGTGG ATCGGCTACA TCTACCCCTA 301 CAGCGGCAGC AGCGACTACA ACCAGAAGTT CAAGAGCAGG GCCACCCTGA 351 CCGTGGACAA CAGCATCAGC ACCGCCTACA TGGAGCTGAG CAGGCTGAGG 401 AGCGACGACA CCGCCGTGTA CTACTGCGCC AGGGGCGGCG ACGCCATGGA 451 CTACTGGGGC CAGGGCACCC TGGTGACCGT GAGCAGCGGT GAGTCGTACG 501 CTAGCAAGCT TTCTGGGGCA GGCCGGGCCT GACTTTGGCT GGGGGCAGGG 551 AGGGGGCTAA GGTGACGCAG GTGGCGCCAG CCAGGTGCAC ACCCAATGCC 601 CATGAGCCCA GACACTGGAC CCTGCATGGA CCATCGCGGA TAGACAAGAA 651 CCGAGGGGCC TCTGCGCCCT GGGCCCAGCT CTGTCCCACA CCGCGGTCAC 701 ATGGCACCAC CTCTCTTGCA GCTTCCACCA AGGGCCCATC CGTCTTCCCC CTGGCGCCCT 751 GCTCCAGGAG CACCTCCGAG AGCACAGCCG CCCTGGGCTG

801 ccTGGTCAAG GACTACTTCC CCGAΆCCGGT GACGGTGTCG TGGAΆCTCAG

851 GCGCCCTGAC CAGCGGCGTG CACACCTTCC CGGCTGTCCT ACAGTCCTCA 901 GGACTCTACT CCCTCAGCAG CGTGGTGACC GTGCCCTCCA GCAGCTTGGG

951 cACGAAGAcc TACACCTGCA ACGTΆGATCA CAAGCCCAGC AACACCAAGG

1001 TGGACAAGAG AGTTGGTGAG AGGCCAGCAC AGGGAGGGAG GGTGTCTGCT 1051 GGAAGCCAGG CTCAGCCCTC CTGCCTGGAC GCACCCCGGC TGTGCAGCCC 1101 CAGCCCAGGG CAGCAAGGCA TGCCCCATCT GTCTCCTCAC CCGGAGGCCT 1151 CTGACCACCC CACTCATGCT CAGGGAGAGG GTCTTCTGGA TTTTTCCACC 1201 AGGCTCCGGG CAGCCACAGG CTGGATGCCC CTACCCCAGG CCCTGCGCAT 1251 ACAGGGGCAG GTGCTGCGCT CAGACCTGCC AAGAGCCATA TCCGGGAGGA 1301 CCCTGCCCCT GACCTAAGCC CACCCCAAAG GCCAAACTCT CCACTCCCTC 1351 AGCTCAGACA CCTTCTCTCC TCCCAGATCT GAGTAACTCC CAATCTTCTC 1401 TCTGCAGAGT CCAAATATGG TCCCCCATGC CCATCATGCC CAGGTAAGCC 1451 AACCCAGGCC TCGCCCTCCA GCTCAAGGCG GGACAGGTGC CCTAGAGTAG 1501 CCTGCATCCA GGGACAGGCC CCAGCCGGGT GCTGACGCAT CCACCTCCAT 1551 CTCTTCCTCA GCACCTGAGT TCCTGGGGGG ACCATCAGTC TTCCTGTTCC 1601 CCCCAAAACC CAAGGACACT CTCATGATCT CCCGGACCCC TGAGGTCACG 1651 TGCGTGGTGG TGGACGTGAG CCAGGAAGAC CCCGAGGTCC AGTTCAACTG

1701 GTACGTGGAT GGCGTGGAGG TGCATAATGC CAΆGACAAAG CCGCGGGAGG

1751 AGCAGTTCAA CAGCACGTAC CGTGTGGTCA GCGTCCTCAC CGTCCTGCAC

1801 CAGGACTGGC TGAACGGCAA GGAGTACAAG TGCAΆGGTCT CCAACAAAGG

1851 CCTCCCGTCC TCCATCGAGA AAACCATCTC CAAAGCCAAA GGTGGGACCC 1901 ACGGGGTGCG AGGGCCACAT GGACAGAGGT CAGCTCGGCC CACCCTCTGC 1951 CCTGGGAGTG ACCGCTGTGC CAACCTCTGT CCCTACAGGG CAGCCCCGAG

2001 AGCCACAGGT GTACACCCTG CCCCCATCCC AGGAGGAGAT GACCAAGAΆC

2051 CAGGTCAGCC TGACCTGCCT GGTCAAAGGC TTCTACCCCA GCGACATCGC 2101 CGTGGAGTGG GAGAGCAATG GGCAGCCGGA GAACAACTAC AAGACCACGC 2151 CTCCCGTGCT GGACTCCGAC GGCTCCTTCT TCCTCTACAG CAGGCTAACC 2201 GTGGACAAGA GCAGGTGGCA GGAGGGGAAT GTCTTCTCAT GCTCCGTGAT 2251 GCATGAGGCT CTGCACAACC ACTACACACA GAAGAGCCTC TCCCTGTCTC

2301 TGGGTAAΆΓG A TAB 13

TGN2122 / TGN2422. C-kappa LC amino acid sequence

Seq. 59: PiQMTQS PS SLSAS _VGDRVT ITCGASEN iχGAJLNWχQRKPGKA

PKLLIYGATNLADGVPSRFSGSGSGRDYTLTISSLQPEDFATYFCQNILGTWTFGGGTK_VEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Seq. 60:

MGWSCIILFLVATATGVHS = Leader peptide, not part of the mature LC

XXXX = variable segment XXXX = constant segment

CDRs

Here: Kabat = AbM

GASENIYGALN - Seq. 54 GATNLAD - Seq. 55 DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQRKPGKAPKLLIYGATNLADGVPSRF

QNILGTWT - Seq. 56 SGSGSGRDYTLTISSLQPEDFATYFCQNILGTWTFGGGTKVEIK - Seq. 58

GASENIYGALNX ₁₅ GATNLADX ₃₂ QNILGTWT - Seq. 57

TAB 14

TGN2122 / TGN2422. C-kappa LC nucleotide sequence (SEQ ID NO: 63)

1 ATGGGATGGA GCTGTATCAT CCTCTTCTTG GTAGCAACAG CTACAGGTAA 51 GGGGCTCACA GTAGCAGGCT TGAGGTCTGG ACATATATAT GGGTGACAAT 101 GACATCCACT TTGCCTTTCT CTCCACAGGT GTGCATTCCG ACATCCAGAT 151 GACCCAGAGC CCCAGCAGCC TGAGCGCCAG CGTGGGCGAC AGGGTGACCA 201 TCACCTGCGG CGCCAGCGAG AACATCTACG GCGCCCTGAA CTGGTACCAG 251 AGGAÄGCCCG GCAAGGCCCC CAAGCTGCTG ATCTACGGCG CCACCAACCT 301 GGCCGACGGC GTGCCCAGCA GGTTCAGCGG CAGCGGCAGC GGCAGGGACT 351 ACACCCTGAC CATCAGCAGC CTGCAGCCCG AGGACTTCGC CACCTACTTC 401 TGCCAGAACA TCCTGGGCAC CTGGACCTTC GGCGGCGGCA CCAAGGTGGA 451 GATCAAGCGT GAGTCGTACG CTAGCAAGCT TGATATCGAA TTCTAAACTC 501 TGAGGGGGTC GGATGACGTG GCCATTCTTT GCCTAAAGCA TTGAGTTTAC 551 TGCAAGGTCA GAAAAGCATG CAAAGCCCTC AGAATGGCTG CAAAGAGCTC 601 CAACAAAACA ATTTAGAACT TTATTAAGGA ATAGGGGGAA GCTAGGAAGA 651 AACTCAAAAC ATCAAGATTT TAAATACGCT TCTTGGTCTC CTTGCTATAA

701 TTATCTGGGA TAAGCATGCT GTTTTCTGTC TGTCCCTAAC ATGCCCTGTG 751 ATTATCCGCA AACAACACAC CCAAGGGCAG AACTTTGTTA CTTAAACACC 801 ATCCTGTTTG CTTCTTTCCT CAGGAACTGT GGCTGCACCA TCTGTCTTCA

851 TCTTCCCGCC ATCTGATGAG CAGTTGAAAT CTGGAACTGC CTCTGTTGTG 901 TGCCTGCTGA ATAACTTCTA TCCCAGAGAG GCCAAAGTAC AGTGGAAGGT 951 GGATAACGCC CTCCAATCGG GTAACTCCCA GGAGAGTGTC ACAGAGCAGG

1001 ACAGCAAGGA CAGCACCTÄC AGCCTCAGCA GCACCCTGAC GCTGAGCAAA 1051 GCAGACTACG AGAAACACAA AGTCTACGCC TGCGAAGTCA CCCATCAGGG 1101 CCTGAGCTCG CCCGTCACAA AGAGCTTCAA CAGGGGAGAG TGTTAG

Claims

Claims:

An isolated monoclonal antibody specific for CTLA-4 and agonistic, wherein the heavy chain of the antibody contains a sequence selected from the group consisting of (Seq. ID): "22, 23, 24, 25, 26, 27, 28, 29, 30, and 32 ".

The isolated monoclonal antibody of claim 1, wherein the antibody light chain contains a sequence selected from the group consisting of (Seq. ID): "33, 34, 35, 36, 37, and 38".

3. An isolated monoclonal antibody according to claim 1 having a heavy chain containing a sequence according to Seq. ID 27, 28, or 29, preferably containing or consisting of the sequence according to Seq. ID 30 or 32, and with a light chain containing a sequence according to Seq. ID 36 or 37, preferably containing or consisting of a sequence according to Seq. ID 38.

4. An isolated monoclonal antibody specific for CTLA-4 and agonistic, the heavy chain of the antibody containing a sequence selected from the group consisting of (Seq. ID): '43, 44, 45, 46, 47, 48, 49, 50, 51, and 53 ".

The isolated monoclonal antibody of claim 4, wherein the antibody light chain contains a sequence selected from the group consisting of (Seq. ID): "54, 55, 56, 57, 58, and 59".

6. An isolated monoclonal antibody according to claim 4 having a heavy chain containing a sequence according to Seq. ID 48, 49, or 50, preferably containing or consisting of the sequence according to Seq. ID 51 or 53, as well as with a light chain containing a sequence according to Seq. ID 57 or 58, preferably containing or consisting of a sequence according to Seq. ID 59.

7. An isolated monoclonal antibody which is specific for CTLA-4 and agonistic, wherein the antibody does not correspond to a CTLA-4 partial sequence according to SEQ. ID 1 binds.

8. An isolated monoclonal antibody according to claim 7, containing 1, 2, 3, 4, 5, or 6 of the sequences according to SEQ. ID 2 to SEQ. ID 7 or the sequences according to SEQ. ID 8 to SEQ. ID 13.

The isolated monoclonal antibody of claim 7 or 8 which is humanized.

10. An isolated monoclonal antibody according to any one of claims 7 to 9, containing one or both of the sequences in each case according to SEQ. ID 14 and SEQ. ID 15 or SEQ. ID 16 and SEQ. ID 17, or contain one or both of the sequences in each case according to SEQ. ID 18 and SEQ. ID 19 or SEQ. ID 20 and SEQ. ID 21.

11. Isolated protein or peptide containing at least one of the sequences SEQ. ID 2 to 13, in particular one of the sequences SEQ. ID 14 to 17 or SEQ. ID 18 to 21, or one of the sequences Seq. ID 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39, in particular one of the sequences Seq. ID 27, 28, 29, 30, 32, 36, 37, or 38, or one of the sequences Seq. ID 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60, in particular one of the sequences Seq. ID 48, 49, 50, 51, 53, 57, 58, or 59, or consisting of one of said sequences.

12. An isolated nucleic acid encoding a protein or peptide according to claim 11 or a light chain and / or heavy chain of an antibody according to any one of claims 1 to 10, in particular containing or consisting of a sequence Seq. ID 61, 62, or 63.

13. Isolated vector containing at least one nucleic acid according to claim 12.

14. An isolated cell which is transfected with a vector according to claim 13.

15. A pharmaceutical composition comprising a monoclonal antibody according to any one of claims 1 to 10 and / or a protein or peptide according to claim 11, and optionally at least one physiologically acceptable carrier and / or excipient.

16. Use of a monoclonal antibody according to any one of claims 1 to 10 and / or a protein or peptide according to claim 11 for the preparation of a pharmaceutical composition for the prophylaxis and / or therapy of a disease or condition selected from the group consisting of 'rheumatoid arthritis, Type I Diabetic, multiple sclerosis, systemic lupus erythematosus, psoriasis, ulcerative colitis, Crohn's disease, allergies, rejection of allograft organ transplant, in particular organ transplants of the following organs: heart, kidney, liver,

Pancreas, lung, bone marrow, and graft-versus-host disease.

17. A process for producing a monoclonal antibody according to any one of claims 1 to 10, wherein a nucleic acid according to claim 12 is introduced into a vector, wherein by means of the vector a cell is transfected, wherein the transfected cell is cultured, wherein a supernatant of the cultured Cells are separated or wherein the cultured cell is lysed and the lysate is recovered, and wherein the separated supernatant or the lysate, the monoclonal antibodies are separated.

18. A process for the preparation of a pharmaceutical composition according to claim 15, wherein the monoclonal antibody and / or the protein or peptide is mixed in physiologically effective dose with the at least one physiologically acceptable carrier and / or excipient and prepared to a defined dosage form.