RU2804490C2 - New antibody compositions for cancer immunotherapy - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: following is proposed: antigen-binding polypeptide that binds to a human PD-L1 epitope containing a heavy chain variable domain and a light chain variable domain, and their respective sequences essentially consist of pairs of sequences selected from a specific group, a nucleic acid molecule encoding an antigen-binding polypeptide, a pharmaceutical composition for use in inhibiting the biological activity of PD-L1, as well as a method of therapeutic treatment of a subject.

EFFECT: invention is used to the treatment of cancer containing a portion of tumor cells expressing a detectable amount of PD-L1.

12 cl, 28 dwg, 27 ex

Description

[0001] This application claims priority and benefit from co-pending U.S. Provisional Patent Application No. 62/720015, filed August 20, 2018, which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

[0002] The present invention relates to antigen-binding polypeptides that bind human PD-L1, pharmaceutical compositions and uses thereof. Aspects of the invention also relate to an expression system producing such antigen-binding polypeptides or antibodies. The antigen binding polypeptides or pharmaceutical compositions described herein are suitable for treating a subject in need thereof of a pathological condition such as mammalian cancer, infection, and the like.

BACKGROUND OF THE ART

[0003] Immune cells have costimulatory and inhibitory receptors on their surface that interact with membrane-bound and soluble ligands. These receptors serve to regulate the activity, duration and type of immune response by changing the thresholds and duration of activation or inhibition of the immune cell. They are often collectively referred to as immune checkpoints. Many of these checkpoint molecules are members of the B7 superfamily of molecules or the tumor necrosis factor (TNF) superfamily of molecules.

[0004] The B7 family includes both inhibitory and stimulatory coreceptors. For example, on the one hand, ligation of programmed death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) with their corresponding ligands (PD-L1, PD-L2 and B7-1, B7-2, respectively) leads to suppression of activation or generation of regulatory T cells, anergy, exhaustion and apoptosis. On the other hand, ligation of the cluster of differentiation (CD28) and inducible T-cell costimulator (ICOS) receptors with their corresponding ligands leads to increased proliferation and cytokine production. In contrast, the TNF co-stimulatory receptor family includes only stimulatory molecules such as OX40, 4-1BB, CD40, CD27 and their ligands that promote proliferation and differentiation of effector function. In addition, there are other coreceptors that also belong to any of these families, for example, Tim-3, LAG-3, Ceacam-1, etc.

[0005] Over the past couple of decades, it has become clear that many types of cancer create an immunosuppressive environment within the tumor through a variety of mechanisms. The key is the ectopic expression of an inhibitory immune checkpoint ligand (namely PDL1), which suppresses intratumoral T cells. There is also growing evidence that blocking this tumor-mediated immune suppression can derepress intratumoral T cells and allow them to kill the tumor. (Adachi K, Tamada K. Cancer Sci. 2015;106(8):945-50; Rafiq S, et al., Nat Biotechnol. 2018 Aug 13; Hargadon KM, et al., Int Immunopharmacol. 2018;62:29 -39). Blocking can be accomplished using an antibody or in a variety of other ways. This differs from traditional anti-cancer antibody treatment, where the antibody binds to the cancer cell and utilizes complement-dependent cytotoxicity (CDC) as well as antibody-dependent cellular cytotoxicity (ADCC) to directly kill tumor cells.

[0006] CTLA-4 antibodies were the first of a class of immune checkpoint blockade immunotherapies to receive FDA approval. Other targets of blockade, such as PD1 and related molecules, offer greater and different opportunities to enhance antitumor immunity in the clinical setting.

BRIEF DESCRIPTION OF THE INVENTION

[0007] The present invention provides antigen binding polypeptides that bind PD-L1 (or, interchangeably, "anti-PD-L1 polypeptide(s)", "PD-L1 binding polypeptides"), preferably human PD-L1; the polypeptide has one or both of the following characteristics: (a) binds to PD-L1 and inhibits its ability to interact with PD1; and (b) has an isotype or constant region that can trigger ADCC and/or CCZ. The resulting antibody can kill tumor cells in two synergistic ways - through T-cell derepression and direct cytotoxicity. The polypeptides of the present invention can be used to treat tumors alone or in combination with: (a) antibodies targeting other immunosuppressive pathways; (b) chemotherapy or radiation therapy; (c) other mechanisms for blocking immunosuppressive pathways, such as aptamers or RNA interference; or (d) other immunotherapeutic agents, such as cytokines, targeted therapies, etc.

[0008] According to one aspect of the present invention, there is provided an antigen-binding polypeptide, e.g., an antibody, fragment, derivative or analog thereof, that is of the IgG1 isotype and binds to a PD-L1 epitope, preferably with a binding affinity of at least 10 ^-6 M, and having a heavy chain variable domain sequence “consisting essentially of” amino acid sequences selected from the group consisting of SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:14, SEQ ID NO: 18, SEQ ID NO:22, SEQ ID NO:26, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:38, SEQ ID NO:42, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:54, SEQ ID NO:58, SEQ ID NO:62, SEQ ID NO:66, SEQ ID NO:70, SEQ ID NO:74, SEQ ID NO:78, SEQ ID NO:82, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:94, SEQ ID NO:98, SEQ ID NO:102, SEQ ID NO:106 and combinations thereof, which means, as used herein, a sequence of at least 80 %, or more preferably, 85%, 90%, 95% or even 100% identical to the specified amino acid sequences; and a light chain variable domain sequence consisting essentially of sequences that are at least 80%, or more preferably 85%, 90%, 95%, or even 100% identical to amino acid sequences selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:20, SEQ ID NO:24, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO :36, SEQ ID NO:40, SEQ ID NO:44, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:56, SEQ ID NO:60, SEQ ID NO:64, SEQ ID NO:68 , SEQ ID NO:72, SEQ ID NO: 76, SEQ ID NO:80, SEQ ID NO:84, SEQ ID NO:88, SEQ ID NO:92, SEQ ID NO:96, SEQ ID NO:100, SEQ ID NO:104, SEQ ID NO:108, and combinations thereof.

[0009] In preferred embodiments, the antigen-binding polypeptide or antigen-binding antibody of the present invention comprises a pair of a heavy chain variable region and a light chain variable region, wherein their respective sequences consist essentially of the following pairs: (a) SEQ ID NO:18 and SEQ ID NO :20; (b) SEQ ID NO:42 and SEQ ID NO:44; or (c) SEQ ID NO:34 and SEQ ID NO:36.

[00010] In other preferred embodiments, the antigen binding polypeptide or antigen binding antibody of the present invention includes a pair of a heavy chain variable region and a light chain variable region, wherein their respective sequences consist essentially of the following pairs: (a) SEQ ID NO:22 and SEQ ID NO:24; (b) SEQ ID NO:2 and SEQ ID NO:4; (c) SEQ ID NO:62 and SEQ ID NO:64; or (d) SEQ ID NO:82 and SEQ ID NO:84.

[00011] In other preferred embodiments, the antigen binding polypeptide or antigen binding antibody of the present invention includes a pair of a heavy chain variable region and a light chain variable region, wherein their respective sequences consist essentially of the following pairs: (a) SEQ ID NO:70 and SEQ ID NO:72; (b) SEQ ID NO:50 and SEQ ID NO:52; (c) SEQ ID NO:102 and SEQ ID NO:104; or (d) SEQ ID NO:30 and SEQ ID NO:32.

[00012] In other preferred embodiments, the antigen binding polypeptide or antigen binding antibody of the present invention includes a pair of a heavy chain variable region and a light chain variable region, wherein their respective sequences consist essentially of the following pairs: (a) SEQ ID NO:6 and SEQ ID NO:8; (b) SEQ ID NO:10 and SEQ ID NO:12; (c) SEQ ID NO:14 and SEQ ID NO:16; (d) SEQ ID NO:26 and SEQ ID NO:28; (e) SEQ ID NO:38 and SEQ ID NO:40; (f) SEQ ID NO:46 and SEQ ID NO:48; (g) SEQ ID NO:54 and SEQ ID NO:56; or (h) SEQ ID NO:58 and SEQ ID NO:60.

[00013] In other preferred embodiments, the antigen binding polypeptide or antigen binding antibody of the present invention includes a pair of a heavy chain variable region and a light chain variable region, wherein their respective sequences consist essentially of the following pairs: (a) SEQ ID NO:66 and SEQ ID NO:68; (b) SEQ ID NO:74 and SEQ ID NO:76; (c) SEQ ID NO:78 and SEQ ID NO:80; (d) SEQ ID NO:86 and SEQ ID NO:88; (e) SEQ ID NO:90 and SEQ ID NO:92; (f) SEQ ID NO:94 and SEQ ID NO:96; (g) SEQ ID NO:98 and SEQ ID NO:100; or (h) SEQ ID NO:106 and SEQ ID NO:108.

[00014] Preferably, said antigen binding polypeptide is entirely human or otherwise humanized. In a preferred embodiment, said antigen binding polypeptide further comprises a human constant region. In one feature, said human constant region is IgG1. In some embodiment, the antibody of the present invention further includes a second pair of heavy and light chain variable regions that, for example, is substantially identical to the first pair.

[00015] In a preferred embodiment, binding of the anti-PD-L1 polypeptide to PD-L1 blocks the interaction of PD-L1 with PD1. This may be due to either the PD-L1 binding epitope being located at or adjacent to the PD1 interaction interface or an allosteric change in the conformation of the PD1 interaction interface.

[00016] According to another aspect of the present invention, nucleic acid molecules are provided that encode the aforementioned polypeptides. The nucleic acid molecule may be a DNA or RNA molecule. In preferred embodiments, the nucleic acid molecule is a DNA molecule encoding a heavy chain variable region and a light chain variable region of an antigen-binding polypeptide or antibody of the present invention, wherein the DNA sequences consist essentially of, respectively, the following pairs: (a) SEQ ID NO:17 and SEQ ID NO:19; (b) SEQ ID NO:33 and SEQ ID NO:35; (c) SEQ ID NO:41 and SEQ ID NO:43.

[00017] In other preferred embodiments, the nucleic acid molecule is a DNA molecule encoding a heavy chain variable region and a light chain variable region of an antigen binding polypeptide or antibody of the present invention, wherein the DNA sequences consist essentially of, respectively, the following pairs of (a) SEQ ID NO:21 and SEQ ID NO:23; (b) SEQ ID NO:1 and SEQ ID NO:3; (c) SEQ ID NO:61 and SEQ ID NO:63; or (d) SEQ ID NO:81 and SEQ ID NO:83.

[00018] In other preferred embodiments, the nucleic acid molecule is a DNA molecule encoding a heavy chain variable region and a light chain variable region of an antigen binding polypeptide or antibody of the present invention, wherein the DNA sequences consist essentially of, respectively, the following pairs: (a) SEQ ID NO:69 and SEQ ID NO:71; (b) SEQ ID NO:49 and SEQ ID NO:51; (c) SEQ ID NO:101 and SEQ ID NO:103; or (d) SEQ ID NO:29 and SEQ ID NO:31.

[00019] In other preferred embodiments, the nucleic acid molecule is a DNA molecule encoding a heavy chain variable region and a light chain variable region of an antigen-binding polypeptide or antibody of the present invention, wherein the DNA sequences consist essentially of, respectively, the following pairs: (a) SEQ ID NO:5 and SEQ ID NO:7; (b) SEQ ID NO:9 and SEQ ID NO:11; (c) SEQ ID NO:13 and SEQ ID NO:15; (d) SEQ ID NO:25 and SEQ ID NO:27; (e) SEQ ID NO:37 and SEQ ID NO:39; (f) SEQ ID NO:45 and SEQ ID NO:47; (g) SEQ ID NO:53 and SEQ ID NO:55; or (h) SEQ ID NO:57 and SEQ ID NO:59.

[00020] In other preferred embodiments, the nucleic acid molecule is a DNA molecule encoding a heavy chain variable region and a light chain variable region of an antigen binding polypeptide or antibody of the present invention, wherein the DNA sequences consist essentially of, respectively, the following pairs: (a) SEQ ID NO:65 and SEQ ID NO:67; (b) SEQ ID NO:73 and SEQ ID NO:75; (c) SEQ ID NO:77 and SEQ ID NO:79; (d) SEQ ID NO:85 and SEQ ID NO:87; (e) SEQ ID NO:89 and SEQ ID NO:91; (f) SEQ ID NO:93 and SEQ ID NO:95; (g) SEQ ID NO:97 and SEQ ID NO:99; or (h) SEQ ID NO:105 and SEQ ID NO:107.

[00021] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising an antigen binding polypeptide, for example, an anti-PD-L1 antibody, fragment, derivative or analog thereof as described herein. The pharmaceutical composition further includes a pharmaceutically acceptable excipient, carrier or diluent.

[00022] According to a similar aspect of the present invention, there is provided a method of therapeutically treating a subject in need thereof for a medical condition, comprising administering to said subject a therapeutically effective amount of an anti-PD-L1 polypeptide or antibody disclosed herein. The method may further comprise the step of administering a second, different therapeutic antibody against at least one cell surface antigen indicative of the condition. The condition being treated may be a mammalian cancer, an infection, or the like. In various embodiments, the anti-PD-L1 polypeptide may be an antibody, an antibody fragment, an antibody derivative, or an antibody analogue.

[00023] Preferably, the range of mammalian cancers to be treated is selected from the group consisting of: ovarian cancer, colon cancer, breast cancer, lung cancer, myelomas, CNS tumors of neuroblastic origin, monocytic leukemias, B-cell leukemias, T-cell leukemia, B-cell lymphoma, T-cell lymphoma, mast cell tumors, melanoma, bladder cancer, gastric cancer, liver cancer, urothelial carcinoma, cutaneous carcinoma, kidney cancer, head and neck cancer, pancreatic cancer and their combinations. More broadly, any cancer in which at least a significant portion of the tumor cells express a detectable amount of PD-L1 is contemplated to be a target for treatment with the composition of the present invention.

[00024] According to another aspect of the present invention, there is provided a method of prophylactically treating a subject in need thereof with similar conditions, comprising administering to said subject a prophylactically effective amount of a pharmaceutical composition of the present invention. The method may further include the step of administering a vaccine against the condition. In one embodiment, said condition is cancer.

[00025] According to a further aspect of the present invention, there is provided a mammalian expression system that produces an antigen binding polypeptide, eg, an antibody, fragment, derivative or analog thereof, that binds to a PD-L1 epitope as described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

[00026] In FIG. 1 schematically depicts screening for antigen-binding polypeptides using solid-phase phage panning technologies, particularly using indirect coating of immunosorption tubes with test proteins, according to an embodiment of the present invention.

[00027] In FIG. 2 schematically depicts the screening of antigen-binding polypeptides using solid-phase phage panning technologies, in particular using direct coating of immunoadsorption tubes with test proteins, according to an embodiment of the present invention.

[00028] FIG. 3 is a table of data that characterizes the hPDL1 binding ability of representative single chain variable fragments (scfv) obtained in an embodiment of the present invention in an indirect ELISA binding assay. "OK" means negative control.

[00029] FIG. 4 is a table of data that characterizes the hPDL1 binding ability of representative single chain variable fragments (scfv) obtained in an embodiment of the present invention in a FACS binding assay. “PC” indicates positive control using APC-conjugated anti-hPDL1 antibody-stained hPDL1/293T cells (10 μg/ml). “OK” indicates negative control with unstained hPDL1/293T cells.

[00030] FIG. 5 is a table of data that characterizes the ability to block the interaction between hPD1 and hPDL1 of various single chain variable fragments (scfvs) obtained in an embodiment of the present invention in a receptor blocking assay (hPDL1-coated plates). “PC” indicates positive control with added biotin-hPD1-Fc. “OK” indicates a negative control in which only buffer was added.

[00031] FIG. 6 is a table of data that characterizes the ability to block the interaction between hPD1 and hPDL1 of various single chain fragment variable (scfv) obtained in an embodiment of the present invention in a receptor blocking assay (hPD1-coated plates). “PC” indicates positive control with added biotin-hPDL1-Fc. “OK” indicates a negative control in which only buffer was added.

[00032] FIG. 7 illustrates the ability of single chain variable fragments (scfv) produced in an embodiment of the present invention in direct ELISAs to bind hPDL1-Fc, mPDL1-Fc (mouse PDL1) and hIgG1.

[00033] In FIG. 8A and 8B show full-length antibody 4-1E8 characterized by SDS-PAGE (FIG. 8A) and size exclusion chromatography (FIG. 8B).

[00034] In FIG. 9A and 9B show full-length antibody 3-1B11, characterized by SDS-PAGE (Fig. 9A) and size exclusion chromatography (Fig. 9B).

[00035] In FIG. 10A and 10B show full length antibody 3-1E4 characterized by SDS-PAGE (FIG. 10A) and size exclusion chromatography (FIG. 10B).

[00036] In FIG. 11B and 11C show the results of a quantitative binding assay of certain embodiments of a full-length antibody of the present invention to hPDL1 in the ELISA format of FIG. 11A.

[00037] In FIG. 12A and 12B show quantitative FACS results for certain embodiments of a full-length antibody of the present invention where binding to hPDL1-expressing 293T cells (upper panel) and hPDL1-negative 293T cells (lower panel) occurs.

[00038] In FIG. 13B shows the results of a receptor blocking assay for the best antibody candidates of the present invention in RBA 1 format (FIG. 13A): hPDL1-Fc coated and Biotin-hPD1-Fc added.

[00039] In FIG. 14B shows the results of a receptor blocking assay for the best antibody candidates of the present invention in RBA 2 format (FIG. 14A): hPD1-Fc coated and Biotin-hPDL1-Fc added.

[00040] FIG. 15 is a table of data that characterizes various full length antibodies produced in an embodiment of the present invention.

[00041] In FIG. 16A-16D show the PD-L1 affinity of the top candidate antibodies using BIAcore: FIG. 16A is a schematic representation of the BIAcore format used in accordance with an example of the present invention; in fig. 16B lists the PD-L1 affinity testing results of the top candidate antibodies using BIAcore; in fig. 16C shows the response curve of antibody codenamed 4-1E8 in BIAcore affinity testing; and in fig. 16D depicts the response curve of the antibody code-named 3-1B11 in BIAcore affinity testing.

[00042] In FIG. 17A is a schematic diagram of an epitope binning format used in accordance with an example of the present invention. In fig. 17B schematically depicts epitope groups (“bins”) for the best antibody candidates according to an embodiment of the present invention. In fig. 17C lists the epitope binning matrix for the best candidate antibodies using the format presented in FIG. 17A.

[00043] In FIG. 18A-18D show the binding ability of: controls (FIG. 18A), antibodies of the present invention codenamed "4-1E8" (FIG. 18B), "3-1E4" (FIG. 18C) and "3-1B11" ( Fig. 18D) with 293T cells transfected with the rhesus PDL1-GFP expressing construct (top) and parental 293T cells (bottom) in FACS analyses.

[00044] In FIG. 19A-19D show the binding ability of: controls (FIG. 19A), antibodies of the present invention codenamed "4-1E8" (FIG. 19B), "3-1E4" (FIG. 19C), and "3-1B11" ( Fig. 19D) with 293T cells transfected with the rhesus PDL1 expressing construct (top) and parental 293T cells (bottom) in FACS analyses.

[00045] In FIG. 20 shows representative EC50 data for an IL-2 production experiment according to embodiments of the invention.

[00046] In FIG. 21 shows ADCC activity of an embodiment of the polypeptide codenamed "4-1E8" compared to the commercially available anti-PDL1 antibody atezolizumab.

[00047] In FIG. 22A-22C show the ADCC activity of the polypeptide embodiment codenamed "4-1E8" compared to embodiments codenamed "3-1B11" (FIG. 22A) and "3-1E4" (FIG. 22B); the table summarizes the main data points (Fig. 22C).

[00048] In FIG. 23A, 23B and 23C show three sets of experimental data on the IL-2 production capacity of peripheral blood mononuclear cells (PBMCs) co-cultured with MDA-MB-231 PDL1+ tumor cells in the presence of the best antibodies of the present invention, compared with commercially available ones. antibodies against PDL1.

[00049] In FIG. 24 presents the results of an analysis of the IFN-γ production capacity of CD8 T cells co-cultured with PDL1+ MDA-MB-231 tumor cells in the presence of the best antibodies of the present invention, in comparison with commercially available anti-PDL1 antibodies.

[00050] In FIG. 25A and 25B show the results of a mixed lymphocyte culture reaction for the best antibodies in accordance with embodiments of the invention.

[00051] In FIG. 26A and 26B show the binding specificity of antibodies of the present invention codenamed "4-1E8" (FIG. 26A) and "3-1B11" (FIG. 26B).

[00052] In FIG. 27A and 27B show the ability of antibodies of the present invention E8 (FIG. 27A) and B11 (FIG. 27B) to block the binding of CD80 to cells expressing PD-L1 (curves shaded in gray), compared to CD80 alone ( solid line) and secondary antibody separately (dashed line).

[00053] In FIG. 28 shows a half-life measurement of an antibody embodiment using Tg32 mice.

DETAILED DESCRIPTION OF THE INVENTION

[00054] Unless otherwise indicated, technical terms are used with their generally accepted meanings.

[00055] As used herein, the singular terms may mean one or more. When used herein, the singular terms together with the word “comprising” may mean one or more. As used herein, the term “other” can mean a second or more. Moreover, unless the context otherwise requires, terms in the singular include terms in the plural, and terms in the plural include terms in the singular.

[00056] As used herein, “about” refers to a numerical value, including, for example, whole numbers, fractions, and percentages, whether explicitly stated or not. The term "about" generally refers to a range of numerical values (e.g., +/-5-10% relative to the given value) that would be considered by one of ordinary skill in the art to be equivalent to the given value (e.g., having the same function or result) . In some cases, the term “approximately” may include numerical values rounded to the nearest significant number. Unless otherwise stated, “approximately” represents +/-10% of the stated value(s).

[00057] An "antigen-binding polypeptide" is a polypeptide containing a moiety that binds an antigen. Examples of antigen-binding polypeptides include antibodies, fragments (eg, the antigen-binding portion of an antibody), derivatives, and analogs of antibodies.

[00058] The antigen-binding polypeptide or protein may have, for example, the structure of a naturally occurring antibody (also known as an “immunoglobulin”). Each naturally occurring antibody consists of two identical pairs of polypeptide chains, each pair having one “light” chain (approximately 25 kDa) and one “heavy” chain (approximately 50-70 kDa). The variable regions of each light/heavy chain pair form an antibody binding site such that an intact antibody has two binding sites.

[00059] The variable regions of naturally occurring antibody chains exhibit the same general structure of relatively conserved framework regions (FRs) connected by three hypervariable regions, also called complementarity determining regions, or CDRs. From the N-terminus to the C-terminus, both the light and heavy chains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4 domains. The assignment of amino acids to each domain follows the definitions of Kabat et al. in: Sequences of Proteins of Immunological Interest, ^5th Ed., US Dept. of Health and Human Services, PHS, NIH, NIH Publication no. 91-3242, 1991. Other systems for numbering amino acids in immunoglobulin chains include IMGT (International Information System ImMunoGeneTics; Lefranc et al., Dev. Comp. Immunol. 29:185-203; 2005) and AHo (Honegger and Pluckthun, J. Mol Biol 309(3):657–670; 2001).

[00060] Antibodies can be obtained from sources such as serum or plasma, which contain immunoglobulins with different antigen specificities. If such antibodies are affinity purified, they can be enriched for specific antigen specificity. Such enriched antibody formulations typically consist of less than about 10% antibodies having specific binding activity for a particular antigen. By subjecting these formulations to several affinity purification steps, it is possible to increase the proportion of antibody that has specific antigen binding activity. Antibodies produced in this way are often called "monospecific". Monospecific antibody preparations may consist of approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 99 .9% antibody that has specific binding activity against a particular antigen.

[00061] The term "antibody" or "Ab" (and its plural forms) as used herein broadly refers to any immunoglobulin (Ig) molecule consisting of four polypeptide chains, two heavy (H) chains and two light ( L), or to any functional fragment(s), mutant(s), variant(s), derivative(s) or analogue(s) that retain the essential and specific features of epitope binding by an Ig molecule. Such antibody fragment, mutant, variant, derivative or analogue formats are known in the art and include, but are not limited to, Fab, F(ab'), F(ab') ₂ , Fv, single chain antibodies (scFv), single domain antibodies (sdAb ), complementarity determining region (CDR) fragments, chimeric antibodies, diabodies, tribodies, tetrabodies, and polypeptides that contain at least a portion of an immunoglobulin sufficient to mediate specific binding of the antigen to the polypeptide. Fragments, derivatives and analogues of antibodies can be obtained by recombinant DNA methods, or by enzymatic or chemical cleavage of intact antibodies.

[00062] The Fab fragment is a monovalent fragment having V _L , V _H , C _L and C _H1 domains; the F(ab') ₂ fragment is a bivalent fragment containing two Fab fragments linked by a disulfide bridge in the hinge region; the Fd fragment has _VH and _CH1 domains; the Fv fragment has the V _L and V _H domains of one antibody arm; and the dAb fragment has V _H domains, a V _L domain, or an antigen binding fragment of a V _H or V _L domain (see, for example, US Pat. Nos. 6,846,634; 6,696,245, US Patent Applications Pub. 20/0202512; 2004/0202995; 2004/0038291 ; 2004/0009507; 2003/0039958, and Ward et al., Nature 341:544-546, 1989).

[00063] A single chain antibody (scFv) is an antibody in which the V _L and V _H regions are connected through a linker (e.g., a synthetic sequence of amino acid residues) to form a continuous protein chain, the linker being long enough to allow the protein chain to fold and form monovalent antigen binding site (see, for example, Bird et al., 1988, Science 242:423-26 and Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-83). Diabodies are bivalent antibodies containing two polypeptide chains, where each polypeptide chain includes V _H and V _L domains connected by a linker too short to allow pairing between two domains of the same chain, allowing each domain to pair with a complementary domain of the other polypeptide chain ( see, for example, Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-48, and Poljak et al., 1994, Structure 2:1121-23). If the two polypeptide chains of a diabody are identical, then the diabody resulting from their pairing will have two identical antigen-binding sites. Polypeptide chains having different sequences can be used to produce a diabody with two different antigen-binding sites. Likewise, tribodies and tetrabodies are antibodies containing three and four polypeptide chains, respectively, and forming three and four antigen-binding sites, respectively, which may be the same or different.

[00064] The complementarity determining regions (CDRs) and framework regions (FRs) of a given antibody can be identified using the system described in Kabat et al. higher; Lefranc et al., supra, and/or Honegger and Pluckthun, supra. One or more CDRs may be incorporated into a molecule in a covalent or non-covalent manner, converting it into an antigen-binding protein. An antigen-binding polypeptide may include a CDR region or regions as part of a larger polypeptide chain, may covalently link the CDR region or regions to another polypeptide chain, or may non-covalently include a CDR region or regions. CDRs allow an antigen binding protein to specifically bind to a particular antigen of interest.

[00065] An antigen binding polypeptide may have one or more binding sites. If there is more than one binding site, the binding sites may be identical to each other or may be different. For example, a naturally occurring human immunoglobulin typically has two identical binding sites, whereas a "bispecific" or "bifunctional" antibody has two different binding sites.

[00066] As used herein, the term “human antibody” or “humanized antibody” includes all antibodies that have one or more variable and constant regions derived from human immunoglobulin sequences. In one embodiment, all variable and constant domains are derived from human immunoglobulin (fully human or humanized antibody) sequences. These antibodies can be produced in a variety of ways, including by immunization with the antigen of interest in a mouse that has been genetically modified to express antibodies derived from genes encoding the human heavy and/or light chain. A humanized antibody has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions and/or additions such that the humanized antibody is less likely to elicit an immune response and/or elicits a less severe immune response according to compared to an antibody of a non-human species when administered to a human subject. In one embodiment, certain amino acids in the framework and constant domains of the heavy and/or light chains of an antibody of a non-human species are mutated to form a humanized antibody. In another embodiment, the constant domain(s) from a human antibody are fused to the variable domain(s) from a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human species antibody are altered to reduce the likely immunogenicity of said non-human species antibody when administered to a human subject, wherein the altered amino acid residues are either not critical for immunospecific binding of the antibody with its antigen, or the changes made to the amino acid sequence are conservative changes such that the binding of a humanized antibody to the antigen is not significantly worse than the binding of a non-human antibody to the antigen. Examples of the production of humanized antibodies can be found in US patents No. 6054297, 5886152 and 5877293.

[00067] As used herein, the term “chimeric antibody” refers to an antibody that contains one or more regions of one antibody and one or more regions of at least another antibody. In one embodiment, CDR regions from more than one anti-human PD-L1 antibody are freely combined in various combinations in a chimeric antibody.

[00068] Activated T cells express PD1 on the cell surface. Binding of PD-L1 to PD1 activates PD1 and suppresses PD1 ⁺ T cells. The term “neutralizing antibody” or “inhibitory antibody” as used herein refers to an antibody that blocks PD1 activation when an excess of anti-PD-L1 antibody reduces the level of said activation by at least about 20% in the assay described herein in the Examples. In various embodiments, the antigen binding protein reduces the level of PD1 activation by at least 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, and 99.9%.

[00069] Antibody fragments or analogues can be readily prepared by those skilled in the art by following the principles described herein and using methods known in the art. Preferred amino termini and carboxyl termini of the fragments or analogs are located near the boundaries of the functional domains. Structural and functional domains can be identified by comparing nucleotide and/or amino acid sequence data with publicly available or proprietary sequence databases. Computerized comparison methods can be used to identify sequence motifs or domains of predicted protein conformation that occur in other proteins of known structure and/or function. There are known methods for identifying protein sequences that fit into a known three-dimensional structure. See Bowie et al., 1991, Science 253:164.

[00070] As used herein, an antigen-binding polypeptide “specifically binds” to an antigen (eg, human PD-L1) if it binds to the antigen with a dissociation constant of 100 nanomolar or less.

[00071] As used herein, an “antigen-binding domain,” “antigen-binding region,” or “antigen-binding site” is a region of an antigen-binding protein that contains amino acid residues (or other moieties) that interact with an antigen and contribute to the specificity of the antigen-binding protein and its affinity for antigen. For an antibody to specifically bind to its antigen, it must include at least a portion of at least one of its CDR domains.

[00072] As used herein, an “epitope” is a portion of a molecule that is bound by an antigen-binding protein (eg, an antibody). An epitope may contain non-contiguous parts of the molecule (eg, amino acid residues in a polypeptide that are not contiguous in the primary sequence of the polypeptide, but in terms of the tertiary and quaternary structure of the polypeptide are close enough to each other to be bound by an antigen-binding protein).

[00073] As used herein, the terms “polynucleotide,” “oligonucleotide,” and “nucleic acid” are used interchangeably and include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), DNA analogs, or RNA created using nucleotide analogs (eg, peptide nucleic acids and non-naturally occurring nucleotide analogs), and hybrids thereof. The nucleic acid molecule can be single-stranded or double-stranded. In one embodiment, the nucleic acid molecules of the present invention comprise a contiguous open reading frame encoding an antibody or a fragment, derivative, mutant or variant thereof.

[00074] "Vector" as used herein means a nucleic acid that can be used to introduce another related nucleic acid into a cell. One type of vector is a “plasmid,” which is a linear or circular double-stranded DNA molecule into which additional nucleic acid segments can be ligated. Another type of vector is a viral vector (eg, replication-defective retroviruses, adenoviruses and adeno-associated viruses), whereby additional DNA segments can be introduced into the viral genome. Certain vectors are capable of autonomous replication in the host cell into which they are introduced (eg, bacterial vectors containing a bacterial origin of replication and mammalian episomal vectors). Other vectors (eg, non-episomal mammalian vectors) are integrated into the host cell genome after introduction into the host cell and thus replicate along with the host genome. An "expression vector" is a type of vector that can direct the expression of a selected polynucleotide.

[00075] As used herein, a nucleotide sequence is “operably linked” to a regulatory sequence if the regulatory sequence affects the expression (eg, level, timing, or location of expression) of the nucleotide sequence. A “regulatory sequence” is a nucleic acid that affects the expression (eg, level, timing, or localization of expression) of the nucleic acid to which it is operably linked. The regulatory sequence may, for example, affect the regulated nucleic acid directly or through the action of one or more other molecules (eg, polypeptides that bind to the regulatory sequence and/or nucleic acid). Examples of regulatory sequences include promoters, enhancers, and other expression control elements (eg, polyadenylation signals). Additional examples of regulatory sequences are described, for example, in Goeddel, 1990, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. and Baron et al., 1995, Nucleic Acids Res. 23:3605-06.

[00076] Preferably, the broad range of mammalian cancers to be treated with the compositions of the present invention is selected from the group consisting of: ovarian cancer, colon cancer, breast cancer, lung cancer, myelomas, CNS tumors of neuroblastic origin, monocytic leukemias, B -cell leukemias, T-cell leukemias, B-cell lymphomas, T-cell lymphomas, mast cell tumors, melanoma, bladder cancer, gastric cancer, liver cancer, urothelial carcinoma, cutaneous carcinoma, kidney cancer, head cancer and neck, pancreatic cancer and combinations thereof. More broadly, any cancer in which at least a portion of the tumor cells express significant amounts of PD-L1 can potentially be treated with the composition of the present invention.

[00077] Polypeptides according to the present description can be obtained using any standard methods known in the art. In one example, the polypeptides are produced by recombinant DNA techniques by inserting a nucleic acid sequence (eg, cDNA) encoding the polypeptide into a recombinant expression vector and expressing the DNA sequence under expression-promoting conditions.

[00078] Nucleic acids encoding any of a variety of polypeptides disclosed herein can be chemically synthesized. Codon usage can be tailored to improve expression in the cell. This codon usage will depend on the cell type chosen. Specialized codon usage patterns have been developed for E. coli and other bacteria, as well as for mammalian cells, plant cells, yeast cells, and insect cells. See, for example: Mayfield et al., Proc. Natl. Acad. Sci. USA. 2003 100(2):438-42; Sinclair et al. Protein Expr. Purif. 2002(1):96-105; Connell N D Curr. Opin. Biotechnol. 2001 12(5):446-9; Makrides et al. Microbiol. Rev. 1996 60(3):512-38; and Sharp et al. Yeast. 1991 7(7):657-78.

[00079] General methods for manipulating nucleic acids are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Laboratory Press, 2 ed., 1989, or F. Ausubel et al., Current Protocols in Molecular Biology (Green Publishing and Wiley-Interscience: New York, 1987), with periodic updates incorporated herein document by reference. The DNA encoding the polypeptide is operably linked to suitable transcriptional or translational regulatory elements derived from mammalian, viral or insect genes. Such regulatory elements include a transcriptional promoter, an optional transcriptional control operator sequence, a sequence encoding suitable ribosome binding sites for the mRNA, and sequences that control transcription and translation termination. Additionally, the ability to replicate in the host, usually provided by the origin of replication, and a selectable gene to facilitate recognition of transformants are included.

[00080] The recombinant DNA of the present invention may also include any type of protein tag sequence that may be useful for protein purification. Examples of protein tags include, but are not limited to, a histidine tag, a FLAG tag, a myc tag, an HA tag, or a GST tag. A description of suitable cloning and expression vectors for use with bacterial, fungal, yeast and mammalian host cells can be found in Cloning Vectors: A Laboratory Manual, (Elsevier, N.Y., 1985).

[00081] The expression construct of the present invention is introduced into a host cell using a method appropriate for the host cell. Various methods are known in the art for introducing nucleic acids into host cells, including, but not limited to: electroporation; transfection using calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran or other substances; microparticle bombardment; lipofection; and infection (where the vector is an infectious agent). Suitable host cells include prokaryotic, yeast, mammalian, or bacterial cells.

[00082] The proteins described herein can also be produced using cellular translation systems. For such purposes, the nucleic acids encoding the polypeptide must be modified to allow in vitro transcription to produce mRNA, and to allow cell-free translation of the mRNA in the particular cell-free system used, eukaryotic, such as a mammalian or yeast cell-free translation system, or prokaryotic, such as a cell-free translation system. bacterial translation system.

[00083] PD-L1 binding polypeptides can also be prepared by chemical synthesis (eg, by methods described in: Solid Phase Peptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford, Ill.). Modifications in a protein can also be obtained by chemical synthesis.

[00084] The polypeptides of the present invention can be purified by protein isolation/purification methods well known in the art of protein chemistry. Non-limiting examples include extraction, recrystallization, salting out (eg with ammonium sulfate or sodium sulfate), centrifugation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic interaction chromatography, normal phase chromatography, reverse phase chromatography, gel filtration, size exclusion chromatography , affinity chromatography, electrophoresis, countercurrent distribution, or any combination thereof. After purification, the polypeptide buffer can be replaced with other buffers and/or the polypeptides can be concentrated by any of a variety of methods known in the art, including, but not limited to, filtration and dialysis.

[00085] The purified polypeptide is preferably at least 85% pure, more preferably at least 90% or 95% pure, and most preferably at least 98%. Regardless of the exact purity number, the polypeptide is sufficiently purified for use as a pharmaceutical product.

Post-translational modifications of polypeptides

[00086] In a certain embodiment, the binding polypeptides of the invention may further contain post-translational modifications. Examples of post-translational modifications of proteins include phosphorylation, acetylation, methylation, ADP-ribosylation, ubiquitination, glycosylation, carbonylation, sumoylation, biotinylation, or the addition of a polypeptide side chain or hydrophobic group. As a result, modified soluble polypeptides may contain non-amino acid elements such as lipids, poly- or monosaccharides and phosphates. The preferred form of glycosylation is sialylation, in which one or more sialic acid moieties are conjugated to a polypeptide. Sialic acid moieties improve solubility and increase serum half-life and reduce the potential immunogenicity of the protein. See Raju et al. Biochemistry. 2001 31; 40(30):8868-76. The effect of such non-amino acid elements on the functionality of the polypeptide can be tested for its antagonizing role on PD-L1 or PD-1 function, for example its inhibitory effect on angiogenesis or tumor growth.

[00087] In one embodiment, the modified forms of the subject polypeptides include linking the subject soluble polypeptides to non-protein polymers. In one specific embodiment, said polymer is polyethylene glycol (“PEG”), polypropylene glycol, or polyoxyalkylenes, as set forth in US Pat. No. 4,640,835; No. 4496689; No. 4301144; No. 4670417; No. 4791192 or No. 4179337.

[00088] In one feature, PEGylated variants of the binding polypeptides of the present invention preferably retain at least 25%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% of the biological activity associated with the unmodified protein. In one embodiment, biological activity refers to the ability to bind to PD-L1, which is assessed by KD, k _on or k _off rates. In one specific embodiment, the PEGylated polypeptide binding protein exhibits increased binding to human PD-L1 compared to its non-PEGylated counterpart. In another embodiment, the biological activity relates to blockade of the PD-L1/PD1 interaction.

Therapeutics, vaccines and administration

[00089] The present invention also provides methods for treating conditions or preventing pre-conditions that respond to inhibition of PD-L1 biological activity. Preferred examples are conditions that are characterized by cellular hyperproliferation and prolonged infection. Methods and dosages for administration vary depending on the type of specific polypeptide and the specific condition being treated. Because regulatory authorities require that a protein reagent used as a therapeutic agent be provided in a formulation with acceptably low levels of pyrogens, the therapeutic compositions of the present invention can be distinguished from other formulations by the fact that they are substantially free of pyrogens or at least contain no more than acceptable levels of pyrogens as determined by the appropriate regulatory authority (e.g. FDA in the USA).

[00090] The pharmaceutical compositions of the present invention may include at least one pharmaceutically acceptable diluent, carrier or excipient. The excipients included in the compositions have different purposes depending, for example, on the type of gene construct or effector cells used, and on the route of administration. Examples of commonly used vehicles include, but are not limited to: saline, buffered saline, dextrose, water for injection, glycerol, ethanol and combinations thereof, stabilizing agents, solubilizing agents and surfactants, buffers and preservatives, tonicity control agents, bulking agents agents and lubricants.

[00091] According to another embodiment of the invention, a pharmaceutical composition of the invention is administered to a patient. Examples of routes of administration include, but are not limited to, intravenous injection. Other modalities include, but are not limited to, intratumoral, intradermal, subcutaneous (SC, SC, sub-Q, Hypo), intramuscular (IM), intraperitoneal (IP), intra-arterial, intraosseous, intracardiac, intra-articular (into the joint), intra-articular (into the area of synovial fluid), intracranial, intravertebral and intrathecal (into the cerebrospinal fluid). Any known device suitable for parenteral injection or infusion of formulations can be used to effect such administration. As used herein, the terms “treat” and “treating” have their usual and customary meanings, and include one or more of: blocking, ameliorating, or reducing the severity and/or frequency of a symptom of a disease (e.g., cancer) in a subject, and/or inhibiting growth, the division, spread or proliferation of cancer cells, or the progression of cancer (eg, the appearance of new tumors) in a subject. Treatment means blocking, improving, reducing or inhibiting by approximately 5-100% compared to a subject in which the methods of the present invention have not been practiced. Preferably, the blocking, enhancement, reduction or inhibition is approximately 100%, 99%, 95%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or 5% compared to a subject in which the methods of the present invention have not been practiced.

[00092] The present invention provides a kit comprising one or more containers filled with a number of gene constructs encoding the polypeptides of the present invention with pharmaceutically acceptable excipients. The kit may also include instructions for use. In addition, a notice may be provided with the kit, in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceutical or biological products, reflecting that agency's approval of manufacture, use, or sale for administration to humans.

EXAMPLES

[00093] Antigen binding polypeptide screening using phage display methods:

[00094] Indirect Coating: FIG. 1 - PDL1-binding single chain variable fragments (scFv) were identified by a standard phage display method. Naïve human scFv libraries were generated by PCR-based reconstruction from B cells from 50 healthy donors. Solid phase panning in immunosorption tubes was performed using an hPDL1-Fc fusion protein and an irrelevant Fc fusion protein indirectly immobilized in an immunosorption tube coated with an anti-human Fc IgG antibody. To sort for proteins with strong binding, Fc-binding scFvs were first depleted using irrelevant Fc-fusion proteins, and then unbound phages were selected for binding to the hPDL1-Fc fusion protein. The eluted phages were amplified in bacteria. These steps were repeated 3-4 times; phage titers and variability were determined after the second stage and beyond. Once sequence convergence was observed (steps 3 and 4), individual phage clones were tested for the ability to bind hPDL1 in an ELISA.

[00095] Direct Coating: Conducted by direct coating of Fc proteins onto an immunosorption tube without the use of anti-human Fc antibody (FIG. 2).

[00096] Phage binding ELISA:

[00097] ELISA was performed using the same strategy as for panning. For clones obtained by indirect panning, plates were first coated with anti-human Fc antibody and then with Fc protein. For clones obtained by direct panning, plates were directly coated with Fc protein. In indirect ELISAs, phages were tested for the ability to bind hPDL1-Fc and irrelevant Fc protein (or hIgG1) in parallel assays. Phages that showed little binding to the irrelevant Fc protein and high binding to hPDL1 were selected for further sequencing and secondary screening. The data is presented in Fig. 3. Nonspecific binding of most clones is insignificant (the signal value for the Fc protein (1:10 dilution) is less than 0.2). In direct ELISAs, phages were tested for their ability to bind hPDL1-Fc, mPDL1-Fc (mouse PDL1), and hIgG1 in parallel assays. Phages demonstrated no significant binding to mouse PDL1 by any of the best molecules of the present invention, namely, none of the best molecules showed significant cross-reactivity with mouse PDL1. The data is presented in Fig. 7.

[00098] Sequencing:

[00099] Unique clones were identified by first sequencing the heavy chain CDR3 region. These results were later confirmed by sequencing the full sequence. A small subset of clones contained the same CDR3s but had significant divergences in other parts of their sequences.

[000100] Secondary screening using FACS:

[000101] Phages, phage lysates, or bacterial lysates expressing scFv were tested for the ability to preferentially bind to 293T cells that express hPDL1, but not to parental 293T cells. The mean fluorescence intensity (MFI) ratio was used as a basis for identifying positive clones. The data is presented in Fig. 4. Most of the clones showed a high ratio, which can be identified as positive clones.

[000102] Identification of blockers:

[000103] Phages, phage lysates or bacterial lysates expressing scFv were tested for the ability to block the interaction between hPD1 and hPDL1. Binding assays were performed by loading hPD1-Fc or hPDL1-Fc plates. Biotinylated ligand (hPDL1 or hPD1) binding was detected using streptavidin-HRP by standard methods. Loss of binding in the presence of scFv was used to identify potential blockers. The results are presented in Fig. 5 and 6.

[000104] Preparation and characterization of Fc fusion proteins:

[000105] Because scFvs are relatively unstable, some scFvs have been converted into Fc fusions and expressed in mammalian cells. They were purified on protein A columns and tested for their ability to block the PD1-PDL1 interaction as well as their ability to bind 293T cells expressing PDL1.

[000106] Preparation of full-length antibodies:

[000107] Full length antibody genes were constructed by PCR amplification of the VH and VL regions from individual scFv clones and cloned into appropriate expression vectors using standard methods known to those skilled in the art. Full-length antibody proteins were prepared by transiently transfecting suspension-grown 293T cells and purified on a Protein A column using standard methods known to those skilled in the art.

[000108] Characterization of full-length antibodies:

[000109] Examples of full-length antibodies were characterized by SDS-PAGE and size exclusion chromatography (results shown in Figures 8A, 8B, 9A, 9B, 10A and 10B), and quantified their activity in (a) specific binding of hPDL1 in an ELISA ELISA (results shown in Figures 11B and 11C); (b) specific binding of hPDL1-expressing 293T cells and crude 293T cells (results shown in FIGS. 12A and 12B); and (c) blocking the PD1-PDL1 interaction in both versions of the blocking assay. The data obtained for examples of the best antibody candidates in Format 1 and Format 2 are shown in FIG. 13B and 14B. Results data for 27 embodiments of the antibodies of the present invention are shown in FIG. 15.

[000110] PD-L1 interaction affinity in BIAcore assay:

[000111] The best antibody candidates were tested for affinity for PD-L1 using BIAcore (Figures 16B-16D). Briefly, biotinylated hPDL1 was captured by streptavidin on the surface of the sensor chip. The antibody was run on the chip and reaction parameters were calculated using a one-cycle kinetic method based on the stability of the interaction. KD values were estimated using BIAcore X100 2.0 evaluation software in a bivalent analyte binding model.

[000112] Rh PD-L1 binding in FACS

[000113] (A) 293T cells were transiently transfected with a rhesus PDL1-GFP expression construct. Embodiments 4-1E8, 3-1E4 and 3-1B11 were tested and compared to control. The results are shown in Fig. 18A-18D: All three antibodies bound rhesus PDL1.

[000114] (b) 293T cells were transiently transfected with a rhesus PDL1 expression construct with a rhesus PDL1 expression construct. Embodiments 4-1E8, 3-1E4 and 3-1B11 were tested and compared to control. The results are shown in Fig. 19A-19D: All three antibodies bound rhesus PDL1.

[000115] IL-2 induction and EC50 determination

[000116] Peripheral blood mononuclear cells (PBMCs) were isolated from human blood on a Ficoll gradient followed by erythrocyte lysis using standard protocols. For analysis, RPMI+ medium was prepared as follows: 10% PBS, 1% antibiotic-antimycotic (Gibco), and 1% nonessential amino acids (Gibco) were added to RPMI medium modified ATCC (Gibco). After isolation from blood, PBMCs were resuspended in 10-20 ml RPMI ⁺ and cultured overnight at 37ºC with 5% CO ₂ . PBMCs were then seeded into a 96-well tissue culture plate (Corning) at a concentration of 100,000 PBMCs/96 wells; the final volume in each well was 200 μl. Staphylococcal enterotoxin B (SEB) was added at a concentration of 1 ng/ml, and the best antibody was added at a concentration of 20 μg/ml (for screening) or in a concentration range from 50 μg/ml to 0.003 μg/ml. Cells without SEB (eg, no stimulation) were used as controls; with SEB alone or with SEB and isotype control (eg, baseline).

[000117] After a 76 hour incubation at 37°C with 5% CO ₂ PBMCs were centrifuged at 1200 rpm for 15 minutes at room temperature, the supernatants were collected and stored at -20°C. IL-2 ELISA was performed using a commercial IL-2 ELISA kit (Biolegend or Thermofisher) following the manufacturer's instructions. For ELISA, supernatants were diluted to 1/20 - 1/80. Absorbance was measured using a Spectramax3 M3 microplate reader (Molecular Devices) and data analyzed using Graphpad software. The best candidate antibodies were compared with commercially available anti-PD1 antibodies. The results are shown in Fig. 20. In tumor coculture experiments with MDA-MB-231 cells (see Figures 23A-23C), 4-1E8 consistently performed better than 3-1B11 and 3-1E4 in inactivating IL-2 (see Figure 23A-23C). Fig. 23A-23C) and IFN-γ (see Fig. 24). However, all three antibodies were as effective as or more effective than the commercial anti-PDL1 antibodies atezolizumab (Atezo) and durvalumab (Durva) in similar co-culture experiments with T cells and MDA-MB-231.

[000118] ADCC activity

[000119] As shown in FIG. 21 and 22, all three best antibodies showed strong ADCC activity, while atezolizumab (designed to be ADCC negative) showed no activity. Of the three embodiments, 4-1E8 demonstrated the greatest ADCC activity.

[000120] Mixed lymphocyte culture reaction

[000121] Peripheral blood mononuclear cells (PBMCs) were isolated from human blood on a Ficoll gradient followed by erythrocyte lysis using standard protocols. Cells were cultured in serum-free RPMI 1640 medium for 1 hour at 37°C. Nonadherent cells were removed and the remaining monocytes were cultured in RPMI 1640 supplemented with 5% AB human serum, 2 ng/ml GM-CSF, and 10 ng/ml IL-4 (BD Biosciences). Fresh medium supplemented with cytokines was added every 2-3 days. Mature dendritic cells were induced by adding 20 ng/ml TNF-alpha (BD Biosciences) on day 6 and cultured for 24 hours.

[000122] Dendritic cells were collected, phenotyped, and frozen for later use. CD4 T cells were isolated from PBMCs using magnetic microcarriers (Dynal) according to the manufacturer's instructions. CD4 T cells were cultured in 96-well flat-bottom plates (Costar) along with allogeneic dendritic cells at a ratio of 1:2.5 using RPMI 1640 supplemented with 10% AB human serum. Before addition, dendritic cells were treated with 100 mg/ml mitomycin C (Sigma). T cell proliferation was measured by the CFSE (or similar dye) dilution method. IFN-g release was measured using a commercial IFN-g ELISA kit following the manufacturer's instructions. Absorbance was measured using a Spectramax3 M3 microplate reader (Molecular Devices) and data analyzed using Graphpad software. In these studies, the best candidate antibodies according to embodiments of the present invention showed results comparable to other commercially available anti-PD1 and anti-PDL1 antibodies. Examples of data are presented in FIG. 25A and 25B.

[000123] Binding specificity

[000124] Expi293 cell lines were generated that stably expressed multiple members of the B7 family and their receptors. Anti-PDL1 antibody potential was tested by FACS using fluorescent anti-human IgG antibodies. Data obtained for examples of the best candidate antibodies are shown in FIG. 26A and 26B.

[000125] Block CD80-PDL1 binding

[000126] DLD1 cells engineered to express PDL1 were used to detect biotinylated CD80-Fc binding in the presence or absence of anti-PDL1 antibodies, followed by treatment with fluorescent streptavidin. Data obtained for examples of the best candidate antibodies of the present invention are shown in FIG. 27A and 27B.

[000127] Half-life measurement

[000128] Serum half-life was measured using homozygous male Tg32 mice (B6.Cg-Fcgrttm1Dcr Tg(FCGRT)32Dcr/DcrJ, Jackson Labs). 2 mg/kg antibody was injected intravenously on day 0; blood was drawn on day 1 and at various subsequent time points. Plasma was prepared and antibody titers were measured using a sandwich ELISA assay. Titers were normalized to day 1 titers. Antibody responses were also measured, and samples with high titers were excluded from analysis because they often showed unexpected changes in the ELISA. Data obtained for examples of the best candidate antibodies of the present invention are shown in FIG. 28. The half-life of the different antibodies ranged from 6.9 days (3-1E4, see detailed sequence description in Example 9 below) to 10.5 days (3-1B11, see detailed sequence description in Example 11 below) and 12. 3 days (4-1E8, see detailed sequence description in Example 5 below).

[000129] Polypeptide sequences

[000130] Examples of PD-L1 binding polypeptide sequences of the present invention are given below:

Example 1: Antibody code: 4-1A2

VH

[000131] DNA (SEQ ID NO:1)

CAGGTTCAGCTGGTGCAGTCTGGGACTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTC

TCCTGCAAGGCTTCTGGATACACCTTCACCAGTTATGATATCAACTGGGTGCGACAGGCC

ACTGGACAAGGGCTTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACTAT

GCACAGAAGTTTCAGGGCAGGGTCACCATGACCACAGACACTTCTACGGGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGATTTTTA

TGGGGTTCGGGGAGTTATGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000132] AMINO ACID (SEQ ID NO:2)

QVQLVQSGTEVKKPGASVKVSCKASGYTFTSYDINWVRQATGQGLEWMGWINPNSGGTNY

AQKFQGRVTMTTDTSTGTAYMELRSLRSDDTAVYYCARFLWGSGSYDYWGQGTLVTVSS

VL

[000133] DNA (SEQ ID NO:3)

GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCA

AGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCT

GAAGATTTCGCAACTTACTACTGTCAACAGACTTACACATTCCCGCACACTTTTGCCCAG

GGGACCAACCTGGAGATCAAA

[000134] AMINO ACID (SEQ ID NO:4)

DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYTFPHTFAQGTNLEIK

Example 2: Antibody code: 4-1A12

VH

[000135] DNA (SEQ ID NO:5)

CAAGTCCAGCTGGTACAATCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTC

TCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATCAGCGCTTACAATGGTAACACAAACTAT

GCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAGATTGG

ATACAGCTATGGTTACCCCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000136] AMINO ACID (SEQ ID NO:6)

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY

AQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDWIQLWLPLDYWGQGTLVTVSS

VL

[000137] DNA (SEQ ID NO:7)

GACATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAACAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATGGTGCATCCAGTTTGGAAAGTGGGGTCCCATCA

AGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCT

GAAGATTTTGCAACTTACTACTGTCAACAGAGTCACAGTTCCCCCCTCACTTTCGGCGGA

GGGACCAAGGTGGACATCAAA

[000138] AMINO ACID (SEQ ID NO:8)

DIQLTQSPSSLSSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYGASSLESGVPS

RFSGSGSGTDFTLTISSLQPEDFATYYCQQSHSSPLTFGGGTKVDIK

Example 3: Antibody code: 4-1B9

VH

[00139] DNA (SEQ ID NO:9)

GAAGTGCAGCTGGTGCAGTCTGGGGGAGGCTTGGTCCAGCCTGGGAGGTCCCTGAGACTC

TCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCT

CCAGGCAAGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTAT

GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTAT

CTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAAAGATTTG

ATCCCGTTGCGAGATAGTAGGGGGGGGTACTACTACGGTATGGACGTCTGGGGCCAAGGG

ACCACGGTCACCGTCTCCTCAGGGAGT

[000140] AMINO ACID (SEQ ID NO:10)

EVQLVQSGGGLVQPGRSLRLSCAASGFFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYY

ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLIPLRDSRGGYYYGMDVWGQG

TTVTVSS

VL

[000141] DNA (SEQ ID NO:11)

TCTTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATC

ACATGCCAAGGAGACAGCCTCAGAGACTATTATGCAAGCTGGTACCAGCAGAAGCCAGGA

CAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGAATCCCAGACCGA

TTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGACTCAGGCGGAA

GATGAGGCTGACTATTACTGTAACTCCCGTGACAGCGGTGCTTACCATTATGTCTTCGGA

ACTGGGACCAAGGTCACCGTCCTA

[000142] AMINO ACID

SSELTQDPAVSVALGQTVRITCQGDSLRDYYASWYQQKPGQAPVLVIYGKNNRPSGIPDR

FSGSSSGNTASLTITGTQAEDEADYYCNSRDSGAYHYVFGTGTKVTVL

Example 4: Antibody code: 4-1B12

VH

[000143] DNA (SEQ ID NO:13)

CAAATCCAGCTGGTGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTC

TCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCT

CCAGGCAAGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTAT

GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTAT

CTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAAAGGAAGT

ATTATAGGGGATGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA

[000144] AMINO ACID (SEQ ID NO:14)

QIQLVQSGGGVVQPGRSLRLSCAASGFFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYY

ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGSIIGDGAFDIWGQGTMVTVSS

VL

[000145] DNA (SEQ ID NO:15)

GATATTGTGATGACCCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGAGAGCCGGCCTCC

ATCTCCTGCAGGTCTAGTCAGACCCTCCTGCATAATGGATTCAACTTTTTGGATGGTAC

CTGCAGAAGCCAGGGCAGTCTCCACAACTCCTGATGTATTTGGCCTCTAGCCGGGCCTCC

GGGGTCCCTGACAGGTTCAGTGGCAGTGGATCGGGCACAGATTTCACACTGAAAATCAGC

AGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAGGTACACACTGGCCGTAC

ACTTTTGGCCAGGGGACCAAGCTGGATATCAAA

[000146] AMINO ACID (SEQ ID NO:16)

DIVMTQSPLSLPVTLGEPASISCRSSQTLLHNGFNFLDWYLQKPGQSPQLLMYLASSRAS

GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHWPYTFGQGTKLDIK

Example 5: Antibody code: 4-1E8

VH

[000147] DNA (SEQ ID NO:17)

CAAATCCAGCTGGTACAATCTGGGGCTGAGGTGAAGATGCCTGGGGCCTCAGTGACGATT

TCCTGCGAGGCGTCTGGATACAACTTCATCAGCTACTATATACACTGGGTGCGACAGGCC

CCTGGACAAGGCCTTGAGTGGATGGGATTCGTCGTCCCTAGTGGTGGTGCCGCCAGGCTAC

ACACAGAAGTTCCAGGGCAGACTCACCGTGACCAGGGACACGTCCACGAGCACAGTCTAC

ATGGACCTGAACAGCCTGACATCTGACGACACGGCCGTGTATTACTGTGTGCGAGAAATG

AGTGGTGGCTGGTTTGATTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCG

[000148] AMINO ACID (SEQ ID NO:18)

QIQLVQSGAEVKMPGASVTISCEASGYNFISYIHWVRQAPGQGLEWMGFVVPSGGAAGY

TQKFQGRLTVTRDTSTSTVYMDLNSLTSDDTAVYYCVREMSGGWFDFWGQGTLVTVSS

VL

[000149] DNA (SEQ ID NO:19)

GACATCGTGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCAAGTCAGGGCATTAGAAAATGATTTAGGCTGGTATCAGCAAAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCA

AGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCT

GAAGATGTGGCAGTTTATTACTGTCAGCAATATTATAGTACTCCTCTCACTTTCGGCCCT

GGGACCAAAGTGGATATCAAA

[000150] AMINO ACID (SEQ ID NO:20)

DIVMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGKAPKLLIYAASTLQSGVPS

RFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPLTFGPGTKVDIK

Example 6: Antibody code: 4-1G7

VH

[000151] DNA (SEQ ID NO:21)

GAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATCAGCGCTTACAATGGTAACACAAACTAT

GCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAGCCTCA

CCGGTACAGCAGCCCATATGGTGGGCGGAGTACTGGGGCCAGGGAACCCTGGTCACCGTC

TCCTCA

[000152] AMINO ACID (SEQ ID NO:22)

EVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY

AQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARASPVQQPIWWAEYWGQGTLVTV

SS

VL

[000153] DNA (SEQ ID NO:23)

CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATC

TCCTGCACTGGAACCAGCAGTGACGTTGGTGGTTATAACTATGTCTCCTGGTACCAACAG

CACCCAGGCAAAGCCCCCAAACTCATGATTTCTGATGTCAGTAAGCGGCCCCTCAGGGGTT

TCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTC

CAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATACAAGCAACTACACTTTGGTA

TTCGGCGGAGGGACCAAGCTGACCGTCCTA

[000154] AMINO ACID (SEQ ID NO:24)

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMISDVSKRPSGV

SNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSNYTLVFGGGTKLTVL

Example 7: Antibody code: 4-1H10

VH

[000155] DNA (SEQ ID NO:25)

CAGCTGCAGCTACAGCAGTCCGGAGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTCCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTATAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAAGACACGGCCGTGTATTACTGTGCGAGTCATGGT

CGGGCAGCAGCTGGTAGGTACGCTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTC

TCCTCA

[000156] AMINO ACID (SEQ ID NO:26)

QLQLQQSGAEVKKPGSSVKVSCKAPGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIANY

AQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCASHGRAAAGRYAMDVWGQGTTVTV

SS

VL

[000157] DNA (SEQ ID NO:27)

AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGATTCTCCGGGGAAGACGGTAACCATC

TCCTGCACCCGCAGCAGTGGCAGCATTGCCAGCAACTATGTGCAGTGGTACCAGCAGCGC

CCGGGCAGTGCCCCCACCACTGTGATCTATGACGATAAGCAAAGACCCTCTGGGGTCCCT

GATCGGTTCTCGGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGA

CTGACGACTGAGGACGAGGCTGACTACTACTGTCAGTCCTTTGATGGCAGCAGTGTCATC

TTCGGCGGAGGGACCAAGCTGACCGTCCTG

[000158] AMINO ACID (SEQ ID NO:28)

NFMLTQPHSVSDSPGKTVTISCTRSSGSIASNYVQWYQQRPGSAPTTVIYDDKQRPSGVP

DRFSGSIDSSSNSASLTISGLTTEDEADYYCQSFDGSSVIFGGGTKLTVL

Example 8: Antibody code: 3-1H2

VH

[000159] DNA (SEQ ID NO:29)

CAGGTTCAGCTGGTGCAGTCTGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAAAGGAG

CGTTTCTATGATAGTAGTGGTTATTACGATGCTTTTGATATCTGGGGCCAAGGGACAATG

GTCACCGTCTCTTCA

[000160] AMINO ACID (SEQ ID NO:30)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARKERFYDSSGYYDAFDIWGQGTM

VTVSS

VL

[000161] DNA (SEQ ID NO:31)

CAGTCTGCCCTGACTCAGCCTCGCTCAGTGTCCGGGTCTCCTGGGCAGTCAGTCACCATC

TCCTGCACTGGAACCAGCAATGATGTTGGTGGTTATAACTATGTCTCCTGGTACCAACAG

CACCCAGGCAAAGCCCCCAAACTCATGATTTATGATGTCACTAAGCGGCCCTCAGGGGTC

CCTGATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCGTCTCTGGCCTC

CAGCCTGAGGATGAGGCTGACTATTATTGCGCCTCTTATGGAGGCAGGAACAATTTGCTT

TTTGGCGGAGGGACTCAACTGACCGTCTTA

[000162] AMINO ACID (SEQ ID NO:32)

QSALTQPRSVSGSPGQSVTISCTGTSNDVGGYNYVSWYQQHPGKAPKLMIYDVTKRPSGV

PDRFSGSKSGNTASLTVSGLQPEDEADYYCASYGGRNNLLFGGGTQLTVL

Example 9: Antibody code: 3-1E4

VH

[000163] DNA (SEQ ID NO:33)

CAAATCCAGCTGGTACAATCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCCGGAGGGGGA

GCAGTGGCGGACAATAGTTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000164] AMINO ACID (SEQ ID NO:34)

QIQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAGGGAVADNSYWGQGTLVTVSS

VL

[000165] DNA (SEQ ID NO:35)

GACATCCGGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCAAGTCAGGGCATTAGAAATGATTTAGGCTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTACAAAGTGGGGTCCCATCA

AGGTTCAGCGGCAGTGGATCTGCACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCT

GAAGATTTTGCAACTTATTACTGTCTACAAGATTACAATTACCCTCGAACGTTCGGGCCAA

GGGACCAAGGTGGAAATCAAA

[000166] AMINO ACID (SEQ ID NO:36)

DIRMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGKAPKLLIYAASSLQSGVPS

RFSGSGSGTDFTLTISSLQPEDFATYYCLQDYNYPRTFGQGTKVEIK

Example 10: Antibody code: 3-1A8

VH

[000167] DNA (SEQ ID NO:37)

CAAATCCAGCTGGTACAATCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACGGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGACGGT

TCGTATAGCAGCAGCTGGTACTCGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTC

TCCTCA

[000168] AMINO ACID (SEQ ID NO:38)

QIQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDGSYSSSWYSFDYWGQGTLVTV

SS

VL

[000169] DNA (SEQ ID NO:39)

CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATC

TCCTGCACTGGAACCAGCAGTGACGTCGGTGGTTATAACTATGTCTCCTGGTACCAACAG

CACCCAGGCAAAGCCCCCAAACTCATGATTTATGATGTCAGTAATCGGCCCTCAGGGGTT

TCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTC

CAGGCTGAGGACGAGGCTGATTATTACTGCTCCTCATATGCAGGTGATATTAGTTATGTA

CTGTTCGGCGGCGGGACCAAGCTGACCGTCCTA

[000170] AMINO ACID (SEQ ID NO:40)

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGV

SNRFSGSKSGNTASLTISGLQAEDEADYYCSSYAGDISYVLFGGGTKLTVL

Example 11: Antibody code: 3-1B11

VH

[000171] DNA (SEQ ID NO:41)

GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGAGGGTCCCTGAGACTC

TCCTGTGCAGCCTCTGGATTCACTTTTAGTGACTATGACATGATCTGGGTCCGCCAGGCT

CCAGGCAAGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTAT

GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTAT

CTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAAAGAGTTC

TTTGGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA

[000172] AMINO ACID (SEQ ID NO:42)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYDMIWVRQAPGKGLEWVAVISYDGSNKYY

ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKEFFGAFDIWGQGTMVTVSS

VL

[000173] DNA (SEQ ID NO:43)

TCTTCTGAGCTGACTCAGGACCCTGCTGTGTCGGTGGCCTTGGGACAGACAGTCACGATC

ACATGCCAAGGAGACAGCCTCAATTACTATTATGCAAACTGGTTCCAGCTGAAGCCAGGG

CAGGCCCCTGTACTTGTCCTCTTTGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGA

TTCTCTGGCTCCTACTCGGGAAGCACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAA

GATGACCGTGACTATTACTGTAATTCGCGGGACAGCGGTGGTAATCCTTGGGTGTTCGGC

GGAGGGACCAAGCTGACCGTCCTA

[000174] AMINO ACID (SEQ ID NO:44)

SSELTQDPAVSVALGQTVTITCQGDSLNYYYANWFQLKPGQAPVLVLFGKNNRPSGIPDR

FSGSYSGSTASLTITGAQAEDDADYYCNSRDSGGNPWVFGGGTKLTVL

Example 12: Antibody code: 4-1F3

VH

[000175] DNA (SEQ ID NO:45)

CAAATCCAGCTGGTACAATCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAAGGATCATCCCTATCCTTGGTATAGCAGACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTAC

ATGGAACTGAGTAGCCTGGGATCTGAGGACACGGCCGTGTATTTTTGTGCGAGAGAGGGG

GGATCCTTTAGGCACTTTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000176] AMINO ACID (SEQ ID NO:46)

QIQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRIIPILGIADY

AQKFQGRVTITADKSTSTAYMELSSLGSEDTAVYFCAREGGSFRHFDFWGQGTLVTVSS

VL

[000177] DNA (SEQ ID NO:47)

CAGCCTGTGCTGACTCAGCCACCTCAGTCTCTGGGGCCCCAGGGCAGAGGGTCACCATC

TCCTGCGCTGGGAGCGACCCCAACATCGGGACAGGTCATGATGTGCACTGGTACCAGCAA

CTTCCAGGAACAGCCCCCAAACTCGTCATCTATGGTAACACCAATCGGCCCCTCAGGGGTC

CCTGAGCGATTCACTGCCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTC

CAGGCTGAGGATGAGGCTGATTATTACTGCCAGGCCTACGACAGGAGCCTGCGTGGTTAT

GTCTTCGGGACTGGGACCAAGGTCACCGTCCTG

[000178] AMINO ACID (SEQ ID NO:48)

QPVLTQPPSVSGAPGQRVTISCAGSDPNIGTGHDVHWYQQLPGTAPKLVIYGNTNRPSGV

PERFTASKSGTSASLAITGLQAEDEADYYCQAYDRSLRGYVFGTGTKVTVL

Example 13: Antibody code: 4-1G5

VH

[000179] DNA (SEQ ID NO:49)

CAAATCCAGCTGGTACAGTCTGGTGCTGAAGTGAAGAAGCCTGGGGCCTCAGTGAAGGTC

TCCTGCAAGACTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATCAGCGCTTACAATGGTAACACAAACTAT

GCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAACTACA

GGTGACGAGTGGCTACGATTGGCTATAAATGACTACTGGGGCCAGGGAACCCTGGTCACC

GTCTCCTCA

[000180] AMINO ACID (SEQ ID NO:50)

QIQLVQSGAEVKKPGASVKVSCKTSGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY

AQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARTTGDEWLRLAINDYWGQGTLVT

VSS

VL

[000181] DNA (SEQ ID NO:51)

GATATTGTGATGACACAGTCTCCCCTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCC

ATCTCCTGCAGGTCTAGTCTGCGCCTCATGCATCCTAATGGACTCAACTATTTGGATTGG

TACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTAATCTTTTTGGGTTCTCAGCGGGCC

TCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATC

AGCAGAGTGGAGGCTGAGGATGTTGGCATTTATTACTGCATGCAAGCTCTAGAACCTCCG

TACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA

[000182] AMINO ACID (SEQ ID NO:52)

DIVMTQSPLSLPVTPGEPASISCRSSLRLMHPNGLNYLDWYLQKPGQSPQLLIFLGSQRA

SGVPDRFSGSGSGTDFTLKISRVEAEDVGIYYCMQALEPPYTFGQGTKLEIK

Example 14: Antibody code: 4-1C9

VH

[000183] DNA (SEQ ID NO:53)

CAGGTCCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGATCCC

GGGTATAGCAGTGGCTGGAAAGATGATGCTTTTGATATCTGGGGCCAAGGGACAATGGTC

ACCGTCTCTTCA

[000184] AMINO ACID (SEQ ID NO:54)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDPGYSSGWKDDAFDIWGQGTMV

TVSS

VL

[000185] DNA (SEQ ID NO:55)

GAAATTGTGATGACACAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGATACAGCCTCC

CTCTCCTGCAGGGCCAGTCAGACTGTTAGCAGCAACTACTTAGCCTGGTACCAACAGAAA

CCTGGCCAGGCTCCCAGGCTCCTCATCTATGATACATCCAACAGGGCCGCTGGCATCCCG

GCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGTAGCCTAGAG

CCTGAAGATTTGCAGTGTATTACTGTCAGCAGTACGGTAGCTCACTCTGGACGTTCGGC

CAAGGGACCAAGGTGGAAATCAAA

[000186] AMINO ACID (SEQ ID NO:56)

EIVMTQSPGTLLSLSPGDTASLSCRASQTVSSNYLAWYQQKPGQAPRLLIYDTSNRAAGIP

ARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYGSSLWTFGQGTKVEIK

Example 15: Antibody Code 11-A4

VH

[000187] DNA (SEQ ID NO:57)

CAGGTGCAGCTGGTGCAGTCTGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGCGGGG

CAGCAGCTGGTAGCCCTTTGGTACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000188] AMINO ACID (SEQ ID NO:58)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAGQQLVALWYYWGQGTLVTVSS

VL

[000189] DNA (SEQ ID NO:59)

CAGTCTGCCCTGACTCAGCCTCCCTCCGCGTCCGGGTCTCGTGGACAGTCAGTCTCCATC

TCCTGCAGTGGAAGTCGCAGTGACATTGGATATTATAACTATGTCTCCTGGTATCAACAA

CACCCAGCAAAGCCCCCAAACTCATCATTTTTGACGTCAATAAGCGGCCCCTCAGGGGTC

CCTGATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCGTCTCTGGCCTC

CAGCCTGAGGATGAGGCTGACTATTATTGCGCCTCTTATGGAGGCAGGAACAATTTGCTT

TTTGGCGGAGGGACTCAACTGACCGTCTTA

[000190] AMINO ACID (SEQ ID NO:60)

QSALTQPPSASGRGQSVSISCSGSRSDIGYYNYVSWYQQHPGKAPKLIIFDVNKRPSGV

PDRFSGSKSGNTASLTVSGLQPEDEADYYCASYGGRNNLLFGGGTQLTVL

Example 16: Antibody code: 21-A1

VH

[000191] DNA (SEQ ID NO:61)

CAGGTGCAACTGCAGGAGTCGGGCCCAGGACTGGTGGAGCCTTCGGAGACCCTGTCCCTC

ACCTGCACTGTCTCTGGTGGCTCCATCAGTAGTTTCTACTGGAGCTGGATCCGGCAGCCC

CCAGGGAAGGGACTGGAGTGGATTGGCTATATCAATTACAGTGGGAGCACCAACTACAAC

CCCTCCCTCAAGAGTCGAGTCACCATATCAGTAGACACGTCCAAGAACCAGTTCTCCCTG

AAGCTGAGCTCTGTGACCGCCGCAGACACGGCTGTGTATTACTGTGCGAGACAGATATTA

TGGTTCGGGGAGTTAAGGTGGTTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCC

TCA

[000192] AMINO ACID (SEQ ID NO:62)

QVQLQESGPGLVEPSETLSLTCTVSGGSISSFYWSWIRQPPGKGLEWIGYINYSGSTNYN

PSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARQILWFGELRWFDPWGQGTLVTVS

S

VL

[000193] DNA (SEQ ID NO:63)

CAGTCTGCCCTGACTCAGCCTCCCTCCGCGTCCGGGTCTCCTGGACAGTCAGTCACCATC

TCCTGCACTGGAACCAGCAGTGACATTGGTGGTTATAACTATGTCTCCTGGTACCAACTG

CGCCCAGGCAAAGCCCCCAAACTCATGATTTATGACGTCACCAAGCGGCCCCTCAGGGGTC

CCTGATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCGTCTCTGGGCTC

CAGGCTGAGGATGAGGCTGATTATTACTGCAGCTCATATGCAGGCAGCAACAATGTGGTA

TTCGGCGGAGGGACCAAGCTGACCGTCCTA

[000194] AMINO ACID (SEQ ID NO:64)

QSALTQPPSASGSPGQSVTISCTGTSSDIGGYNYVSWYQLRPGKAPKLMIYDVTKRPSGV

PDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYAGSNNVVFGGGTKLTVL

Example 17: Antibody code: 21-H12

VH

[000195] DNA (SEQ ID NO:65)

CAAGTCCAGCTGGTACAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAAATCCC

TACGGTTTCAACTGGTTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000196] AMINO ACID (SEQ ID NO:66)

QVQLVQSGAEVKKPGASVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARNPYGFNWFDPWGQGTLVTVSS

VL

[000197] DNA (SEQ ID NO:67)

AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATC

TCCTGCACCCGCAGCAGTGGCAGCATTGCCAGCAACTATGTGCAGTGGTACCAGCAGCGC

CCGGGCAGTGCCCCCACCACTGTGATCTATGAGGATAACCAAAGACCCTCTGGGGTCCCT

GATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCCGGA

CTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGTCTTATGATGGCTTCAATCAGGTG

TTCGGCGGAGGGACCAAGCTGACCGTCCTA

[000198] AMINO ACID (SEQ ID NO:68)

NFMLTQPHSVSESPGKTVTISCTRSSGSIASNYVQWYQQRPGSAPTTVIYEDNQRPSGVP

DRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDGFNQVFGGGTKLTVL

Example 18: Antibody code: 7-D12

VH

[000199] DNA (SEQ ID NO:69)

CAAATGCAGCTGGTACAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAACCGGT

AGTAGTGGTTATGTACGTTGGAGCAACTGGTTCGACCCCTGGGGCCAGGGAACCCTGGTC

ACCGTCTCCTCA

[000200] AMINO ACID (SEQ ID NO:70)

QMQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARTGSSGYVRWSNWFDPWGQGTLV

TVSS

VL

[000201] DNA (SEQ ID NO:71)

GACATCCAGATGACCCAGTCTCCCTCCACCCTGTCTGCATTTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCCAGTGAGAGTATTAGTAGGTGGTTGGCCTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAACTCCTAATCTCTAAGACGTCTAATTTAGAAAGCGGGGTCCCGTCA

AGGTTCAGTGGCGCTGGATCTGGGACAGATTTCACTCTCACCATTAGCAGTCTGCAACCT

GAGGATTTTGCAACTTACTTCTGTCAACAGGGTTCCAAAATGCCTCCGACTTTCGGCGGA

GGGACCAAGGTGGAGATCAAG

[000202] AMINO ACID (SEQ ID NO:72)

DIQMTQSPSTLSAFVGDRVTITCRASESISRWLAWYQQKPGKAPKLLISKTSNLESGVPS

RFSGAGSGTDFTLTISSLQPEDFATYFCQQGSKMPPTFGGGTKVEIK

Example 19: Antibody code: 9-E3

VH

[000203] DNA (SEQ ID NO:73)

CAGGTGCAGCTGGTGCAGTCTGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGGGGCC

TACGGTGGTAACTCCGCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000204] AMINO ACID (SEQ ID NO:74)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGAYGGNSAFDYWGQGTLVTVSS

VL

[000205] DNA (SEQ ID NO:75)

CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCATC

TCCTGCACTGGGAGCAGCTCCAACATCGGGGCAGGTTATGATGTACACTGGTACCAGCAG

CTTCCAGGAACAGCCCCCAAACTCCTCATGTACAGTAATGATCAGCGGCCCCTCAGGGGTC

ACTGAGCGATTCTCTGGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTC

CAGTCTGAAGATGAGGGTGATTACTACTGCCAGTCCTATGACAGAAGCCTGAGAGGTTCG

GTCTTCGGCGGAGGGACCAAGCTGACCGTCCTC

[000206] AMINO ACID (SEQ ID NO:76)

QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLMYSNDQRPSGV

TERFSGSKSGTSASLAISGLQSEDEGDYYCQSYDRSLRGSVFGGGTKLTVL

Example 20: Antibody code: 10-A6

VH

[000207] DNA (SEQ ID NO:77)

GAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTT

TCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATCAGCGCTTACAATGGTAACACAAACTAT

GCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAGATTCC

ATAGCAGCAGCTGGTACTCCGTTCGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC

TCA

[000208] AMINO ACID (SEQ ID NO:78)

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY

AQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDSIAAAGTPFDYWGQGTLVTVS

S

VL

[000209] DNA (SEQ ID NO:79)

AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTCACCATC

TCCTGCACCCGCAGCAGTGGCATCATTGCCAGCAAATATGTGCACTGGTACCAGCAGCGC

CCGGGCAGTGCCCCCACCACTGTGATCTATGAGGATAACCAAAGACCGTCTGGGGTCCCT

GATCGATTCTCTGGCTCCATCGACAACTCCTCCAACTCTGCCTCCCTCACCATCTCTGGA

CTGCAGACTGAGGACGAGGCTGACTACTACTGTCAGTCTCATGACGGCATCAATCAGGTT

TTCGGCGGAGGGACCAAGGTCACCGTCCTA

[000210] AMINO ACID (SEQ ID NO:80)

NFMLTQPHSVSESPGKTVTISCTRSSGIIASKYVHWYQQRPGSAPTTVIYEDNQRPSGVP

DRFSGSIDNSSNSASLTISGLQTEDEADYYCQSHDGINQVFGGGTKVTVL

Example 21: Antibody code: 12-A4

VH

[000211] DNA (SEQ ID NO:81)

GAGGTGCAGCTGGTGGAGTCCCGGGGAGGCTTGGTACAGCCGGGGGGGTCCCTGAGACTC

TCCTGTGTAACTTCTGGATTCAGCTTTAACAACTATGCCATGAACTGGGTCCGCCAGGCT

CCGGGGAAGGGGCTGGAGTGGGTCTCAGCTGTTAGTGGTAGTGGTGGTACCACATACTAC

GCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTTT

GTGCAGATGGCACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAAAGGACTT

TTCCCTACGATTTTTGGAGTAGGAGCAATGTTTGACTACTGGGGCCAGGGAACCCTGGTC

ACCGTCTCCTCA

[000212] AMINO ACID (SEQ ID NO:82)

EVQLVESRGGLVQPGGSLRLSCVTSGFSFNNYAMNWVRQAPGKGLEWVSAVSGSGGTTYY

ADSVKGRFTISRDNSKNTLFVQMDSLRAEDTAVYYCAKGLFPTIFGVGAMFDYWGQGTLV

TVSS

VL

[000213] DNA (SEQ ID NO:83)

TCTTCTGAGCTGACTCAGCCACCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATC

TCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATGCTGTTAACTGGTATCAGCAGCTC

CCAGGAACGGCCCCCAAACTCCTCATCTATGATAATAATCACCGGCCCTCAGGGGTCCCT

GACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCAG

TCTGAGGATGAGGCTGATTATTATTGTGCAGCATGGGATGACACCATTCCTGGTGTGCTA

TTCGCCGGAGGGACCAAGCTGACCGTCCTA

[000214] AMINO ACID (SEQ ID NO:84)

SSELTQPPSASGTPGQRVTISCSGSSSNIGSNAVNWYQQLPGTAPKLLIYDNNHRPSGVP

DRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDTIPGVLFAGGTKLTVL

Example 22: Antibody code: 14-G10

VH

[000215] DNA (SEQ ID NO:85)

GAAGTGCAGCTGGTGGAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGGTGTT

TCTTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA

[000216] AMINO ACID (SEQ ID NO:86)

EVQLVESGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGVSYYYGMDVWGQGTTVTVSS

VL

[000217] DNA (SEQ ID NO:87)

CAGGCTGTGCTGACTCAGCCACCTCGGTGTCCGTGTCCCCAGGACAGACAGCCATCATC

TCCTGTTCTGGACATAAATTGGGTGATAAGTATGTTTCCTGGTATCAACAGCAGCCAGGC

CAGTCCCCTGTGCTGGTCCTCTTTCAGGATACCAAGCGGCCCTCAGGGATCCCTGAGCGA

TTCTCTGGCTCCAACTCTGGGAACACAGCCACTCTGACCATCAGCGCGACCCAGGCTGCG

GATGAGGCTGACTATTACTGTCAGGCGGGGGACACCAAGTCTGTGATCTTCGGCGGCGGG

ACCAAGCTGACCGTCCTA

[000218] AMINO ACID (SEQ ID NO:88)

QAVLTQPPSVSVSPGQTAIISCSGHKLGDKYVSWYQQQPGQSPVLVLFQDTKRPSGIPER

FSGSNSGNTATLTISATQAADEADYYCQAGDTKSVIFGGGTKLTVL

Example 23: Antibody code: 22-A6

VH

[000219] DNA (SEQ ID NO:89)

CAGGTTCAGGTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGGCAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGGATAC

AGCTATGGTTCAGGACACCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000220] AMINO ACID (SEQ ID NO:90)

QVQVVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGYSYGSGHLDYWGQGTLVTVSS

VL

[000221] DNA (SEQ ID NO:91)

GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCAGGCGAGTCAGGACATTAGCAACTATTTAAATTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAATTTGGAAACAGGGGTCCCATCA

AGGTTCAGTGGCAGTGGATCTGGGACAGATTTCGCTCTCACCATCAGCAGTCTCCAACCT

GAAGATTTTGCAACTTATTACTGTCTACAGCATAATAGTTACCCTCGGACTTTTGGCCAG

GGGACCAAGCTGGAGATCAAA

[000222] AMINO ACID (SEQ ID NO:92)

DIQMTQSPSSLSSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPS

RFSGSGSGTDFALTISSLQPEDFATYYCLQHNSYPRTFGQGTKLEIK

Example 24: Antibody code: 35-B1

VH

[000223] DNA (SEQ ID NO:93)

GAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTC

TCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATGAACCCTAACAGTGGTGACACAGCCTAT

ACACAGAACTTCCAGGGCAGAGTCACCATGACCAGGAACCCCTCCATAAGCACAGCCTAC

ATGGAGCTGAGCAACCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGAGGCCGG

GGGTTCGCGGAGAAGCCCCTTGGGTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000224] AMINO ACID (SEQ ID NO:94)

EVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWMNPNNSGDTAY

TQNFQGRVTMTRNPSISTAYMELSNLRSEDTAVYYCARGRGFAEKPLGYWGQGTLVTVSS

VL

[000225] DNA (SEQ ID NO:95)

GATATTGTGATGACTCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGGGAGGGCCACC

ATCAACTGCAAGTCCAGCCAGAGTATTTTATCCAGCTCCAATAATAAGAACTATTTAGCT

TGGTACCAGCAGAAACCAGGTCAGCCTCCTAAGCTGCTCATTTACTGGGCATCTACCCGG

GAATCCGGGGTCCCTGACCGGTTCAGCGGCAGCGGGTCTGGGACAGATTTCACTCTCACC

ATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGCAATATTATAGTACT

CCTCCGACATTCGGCCAAGGGACCAAGGTGGAAATCAAA

[000226] AMINO ACID (SEQ ID NO:96)

DIVMTQSPDSLAVSLGGRATINCKSSQSILSSSNNKNYLAWYQQKPGQPPKLLIYWASTR

ESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPPTFGQGTKVEIK

Example 25: Antibody code: 3-1F4

VH

[000227] DNA (SEQ ID NO:97)

GAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTC

TCCTGCAAGGCTTCTGGAGGCACCTTCAGCAGCTATGCTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTAC

GCACAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACAGCCTAC

ATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGAGGGCCCCT

CGAGGGCAGTGGCTGGTTCACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTC

TCCTCA

[000228] AMINO ACID (SEQ ID NO:98)

EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANY

AQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAPRGQWLVHYFDYWGQGTLVTV

SS

VL

[000229] DNA (SEQ ID NO:99)

GAAATTGTGTTGACGCAGTCTCCAGCCACCCTCTCTTGTCTCCAGGGGAAAGAGCCACC

CTCTCCTGCTGGGCCAGTCAGGATGTTAGCAACTACTTAGCCTGGTACCAACAGAAGCCT

GGCCAGGCTCCCAGGCTCCTCATCTATGATGCATCCAACAGGGCCACTGGCATCCCAGCC

AGGTTCAGCGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCT

GAAGATTTTGCAGTGTATTACTGTCAGCAACGTAGCAACTGGCCTCTCACTTTCGGCGGC

GGGACCAAGGTGGAGCTCAAA

[000230] AMINO ACID (SEQ ID NO:100)

EIVLTQSPATLSLSPGERATLSCWASQDVSNYLAWYQQKPGQAPRLLIYDASNRATGIPA

RFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVELK

Example 26: Antibody code: 4-1B3

VH

[000232] DNA (SEQ ID NO:101)

CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTC

TCCTGCAAGGCTTCTGGTTACACCTTTACCAGCTATGGTATCAGCTGGGTGCGACAGGCC

CCTGGACAAGGGCTTGAGTGGATGGGATGGATCAGCGCTTACAATGGTAACACAAACTAT

GCACAGAAGCTCCAGGGCAGAGTCACCATGACCACAGACACATCCACGAGCACAGCCTAC

ATGGAGCTGAGGAGCCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAGAGTCC

TACTCGTCCGCAGGTATTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA

[000232] AMINO ACID (SEQ ID NO:102)

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY

AQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARESYSSAGIDYWGQGTLVTVSS

VL

[000233] DNA (SEQ ID NO:103)

GATATTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCC

ATCTCCTGCAGGTCTAGTCAGACCCTCCTGCATAGTAATGGATTCAACTATTTGGATTGG

TACCTGCAGAAGCCAGGGCAGTCTCCACAACTCCTGATGTATTTGGGCTCTAGCCGGGCC

TCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCGGGCACAGATTTCACACTGAAAATC

AGCAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAACTCTACAAACTCCT

CCGGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA

[000234] AMINO ACID (SEQ ID NO:104)

DIVMTQSPLSLPVTPGEPASISCRSSQTLLHSNGFNYLDWYLQKPGQSPQLLMYLGSSRA

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQTLQTPPAFGGGTKVEIK

Example 27: Antibody code: 21-G1

VH

[000235] DNA (SEQ ID NO:105)

CAGGTCCAGCTGGTGCAGTCTGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGACGATT

TCCTGCGAGGCGTCTGGATACAACTTCATCAGCTACTATATACACTGGGTGCGACAGGCC

CCTGGACAAGGCCTTGAGTGGATGGGATTCGTCGTCCCTAGTGGTGGTGCCGCCAGGCTAC

ACACAGAAGTTCCAGGGCAGACTCACCGTGACCAGGGACACGTCCACGAGCACAGTCTAC

ATGGACCTGAACAGCCTGACATCTGACGACACGGCCGTGTATTACTGTGTGCGAGAAATG

AGTGGTGGCTGGTTTGATTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCG

[000236] AMINO ACID (SEQ ID NO:106)

QVQLVQSGAEVKKPGASVTISCEASGYNFISYIHWVRQAPGQGLEWMGFVVPSGGAAGY

TQKFQGRLTVTRDTSTSTVYMDLNSLTSDDTAVYYCVREMSGGWFDFWGQGTLVTVSS

VL

[000237] DNA (SEQ ID NO:107)

GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC

ATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCA

AGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCT

GAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCGATCACCTTCGGCCAA

GGGACACGACTGGGAGATTAAA

[000238] AMINO ACID (SEQ ID NO:108)

DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIK

[000239] It will be apparent to those skilled in the art that various modifications and changes may be made to the present invention without departing from the scope or spirit of the invention. Other embodiments of the invention will become apparent to those skilled in the art upon examination of the description and practice of the invention disclosed herein. It is intended that the present description and examples are illustrative only and that the true scope and spirit of the invention are defined by the following claims.

[000240] The text of this application contains references to various publications, patents and/or patent applications for a more complete description of the prior art to which the present invention relates. The contents of these publications, patents and/or patent applications are incorporated herein by reference in their entirety to the same extent as if each independent publication, patent and/or patent application had been specifically and individually identified as being incorporated by reference.

--->

LIST OF SEQUENCES

<110> 1GLOBE BIOMEDICAL CO., LTD./ 1GLOBE BIOMEDICAL CO., LTD.

1GLOBE HEALTH INSTITUTE LLC/ 1GLOBE HEALTH INSTITUTE LLC

LI, Chiang J./ LI, Chan J.

UNNIRAMAN, Shyam/ UNNIRAMAN, Shyam

BADER, Hannah/ BADER, Hannah

JHA, Mithilesh/ JA, Mithilesh

<120> NEW ANTIBODY COMPOSITIONS FOR CANCER IMMUNOTHERAPY

<130>PI19033299

<140> PCT/CN 2019/101659

<141> 2019-08-20

<150> US 62/720015

<151> 2018-08-20

<160> 108

<170> PatentIn, version 3.5

<210> 1

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1A2 VH

<400> 1

caggttcagc tggtgcagtc tgggactgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggata caccttcacc agttatgata tcaactgggt gcgacaggcc 120

actggacaag ggcttgagtg gatgggatgg atcaacccta acagtggtgg cacaaactat 180

gcacagaagt ttcagggcag ggtcaccatg accacagaca cttctacggg cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagatttta 300

tggggttcgg ggagttatga ctactggggc cagggaaccc tggtcaccgt ctcctca 357

<210> 2

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1A2 VH

<400> 2

Gln Val Gln Leu Val Gln Ser Gly Thr 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 Ser Tyr

20 25 30

Asp Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Gly Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Phe Leu Trp Gly Ser Gly Ser Tyr Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 3

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1A2 VL

<400> 3

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagatttcg caacttacta ctgtcaacag acttacacat tcccgcacac ttttgcccag 300

gggaccaacc tggagatcaa a 321

<210> 4

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1A2 VL

<400> 4

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 Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys 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 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 Thr Tyr Thr Phe Pro His

85 90 95

Thr Phe Ala Gln Gly Thr Asn Leu Glu Ile Lys

100 105

<210> 5

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1A12 VH

<400> 5

caagtccagc tggtacaatc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagattgg 300

atacagctat ggttacccct tgactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 6

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1A12 VH

<400> 6

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 Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Trp Ile Gln Leu Trp Leu Pro Leu Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 7

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1A12 VL

<400> 7

gacatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca acagaaacca 120

gggaaagccc ctaagctcct gatctatggt gcatccagtt tggaaagtgg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agtcacagtt cccccctcac tttcggcgga 300

gggaccaagg tggacatcaa a 321

<210> 8

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1A12 VL

<400> 8

Asp Ile Gln Leu 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 Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Ser Leu Glu 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 Ser His Ser Ser Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys

100 105

<210> 9

<211> 387

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B9 VH

<400> 9

gaagtgcagc tggtgcagtc tgggggaggc ttggtccagc ctgggaggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120

ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180

gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gaaagatttg 300

atcccgttgc gagatagtag gggggggtac tactacggta tggacgtctg gggccaaggg 360

accacggtca ccgtctcctc agggagt 387

<210> 10

<211> 127

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B9 VH

<400> 10

Glu Val Gln Leu Val Gln 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 Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala 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 Lys Asp Leu Ile Pro Leu Arg Asp Ser Arg Gly Gly Tyr Tyr Tyr

100 105 110

Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 11

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B9 VL

<400> 11

tcttctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60

acatgccaag gagacagcct cagagactat tatgcaagct ggtaccagca gaagccagga 120

caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcaggaat cccagaccga 180

ttctctggct ccagctcagg aaacacagct tccttgacca tcactgggac tcaggcggaa 240

gatgaggctg actattactg taactcccgt gacagcggtg cttaccatta tgtcttcgga 300

actgggacca aggtcaccgt ccta 324

<210> 12

<211> 108

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B9 VL

<400> 12

Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Asp Tyr Tyr Ala

20 25 30

Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr

35 40 45

Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Thr Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Gly Ala Tyr His

85 90 95

Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105

<210> 13

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B12 VH

<400> 13

caaatccagc tggtgcagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120

ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180

gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gaaaggaagt 300

attatagggg atggtgcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 360

<210> 14

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B12 VH

<400> 14

Gln Ile Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala 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 Lys Gly Ser Ile Ile Gly Asp Gly Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 15

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B12 VL

<400> 15

gatattgtga tgacccagtc tccactctcc ctgcccgtca cccttggaga gccggcctcc 60

atctcctgca ggtctagtca gaccctcctg cataatggat tcaacttttt ggattggtac 120

ctgcagaagc cagggcagtc tccacaactc ctgatgtatt tggcctctag ccgggcctcc 180

ggggtccctg acaggttcag tggcagtgga tcgggcacag atttcacact gaaaatcagc 240

agagtggagg ctgaggatgt tggggtttat tactgcatgc aaggtacaca ctggccgtac 300

acttttggcc aggggaccaa gctggatatc aaa 333

<210> 16

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B12 VL

<400> 16

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Thr Leu Leu His Asn

20 25 30

Gly Phe Asn Phe Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro

35 40 45

Gln Leu Leu Met Tyr Leu Ala Ser Ser Arg Ala Ser Gly Val Pro Asp

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser

65 70 75 80

Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly Thr

85 90 95

His Trp Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Asp Ile Lys

100 105 110

<210> 17

<211> 354

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1E8 VH

<400> 17

caaatccagc tggtacaatc tggggctgag gtgaagatgc ctggggcctc agtgacgatt 60

tcctgcgagg cgtctggata caacttcatc agctactata tacactgggt gcgacaggcc 120

cctggacaag gccttgagtg gatgggattc gtcgtcccta gtggtggtgc cgcaggctac 180

acacagaagt tccagggcag actcaccgtg accagggaca cgtccacgag cacagtctac 240

atggacctga acagcctgac atctgacgac acggccgtgt attactgtgt gcgagaaatg 300

agtggtggct ggtttgattt ctggggccag ggaaccctgg tcaccgtctc ctcg 354

<210> 18

<211> 118

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1E8 VH

<400> 18

Gln Ile Gln Leu Val Gln Ser Gly Ala Glu Val Lys Met Pro Gly Ala

1 5 10 15

Ser Val Thr Ile Ser Cys Glu Ala Ser Gly Tyr Asn Phe Ile Ser Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Phe Val Val Pro Ser Gly Gly Ala Ala Gly Tyr Thr Gln Lys Phe

50 55 60

Gln Gly Arg Leu Thr Val Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Asp Leu Asn Ser Leu Thr Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Arg Glu Met Ser Gly Gly Trp Phe Asp Phe Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 19

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1E8 VL

<400> 19

gacatcgtga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcaaaaacca 120

gggaaagccc ctaagctcct gatctatgct gcatccactt tgcaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcaggct 240

gaagatgtgg cagtttatta ctgtcagcaa tattatagta ctcctctcac tttcggccct 300

gggaccaaag tggatatcaa a 321

<210> 20

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1E8 VL

<400> 20

Asp Ile Val 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 Gly Ile Arg Asn Asp

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Thr Leu Gln 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 Ala

65 70 75 80

Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Leu

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 21

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1G7 VH

<400> 21

gaggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagcctca 300

ccggtacagc agcccatatg gtgggcggag tactggggcc agggaaccct ggtcaccgtc 360

tcctca 366

<210> 22

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1G7 VH

<400> 22

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ala Ser Pro Val Gln Gln Pro Ile Trp Trp Ala Glu Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 23

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1G7 VL

<400> 23

cagtctgccc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc gatcaccatc 60

tcctgcactg gaaccagcag tgacgttggt ggttataact atgtctcctg gtaccaacag 120

cacccaggca aagcccccaa actcatgatt tctgatgtca gtaagcggcc ctcaggggtt 180

tctaatcgct tctctggctc caagtctggc aacacggcct ccctgaccat ctctgggctc 240

caggctgagg acgaggctga ttattactgc agctcatata caagcaacta cactttggta 300

ttcggcggag ggaccaagct gaccgtccta 330

<210> 24

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1G7 VL

<400> 24

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Ser Asp Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Asn

85 90 95

Tyr Thr Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 25

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1H10 VH

<400> 25

cagctgcagc tacagcagtc cggagctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg ctcctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaagg atcatcccta tccttggtat agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaagac acggccgtgt attactgtgc gagtcatggt 300

cgggcagcag ctggtaggta cgctatggac gtctggggcc aagggaccac ggtcaccgtc 360

tcctca 366

<210> 26

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1H10 VH

<400> 26

Gln Leu Gln Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Pro Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr 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 Ser His Gly Arg Ala Ala Ala Gly Arg Tyr Ala Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 27

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1H10 VL

<400> 27

aattttatgc tgactcagcc ccactctgtg tcggattctc cggggaagac ggtaaccatc 60

tcctgcaccc gcagcagtgg cagcattgcc agcaactatg tgcagtggta ccagcagcgc 120

ccgggcagtg cccccaccac tgtgatctat gacgataagc aaagaccctc tggggtccct 180

gatcggttct cgggctccat cgacagctcc tccaactctg cctccctcac catctctgga 240

ctgacgactg aggacgaggc tgactactac tgtcagtcct ttgatggcag cagtgtcatc 300

ttcggcggag ggaccaagct gaccgtcctg 330

<210> 28

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1H10 VL

<400> 28

Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Asp Ser Pro Gly Lys

1 5 10 15

Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ser Ile Ala Ser Asn

20 25 30

Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro Thr Thr Val

35 40 45

Ile Tyr Asp Asp Lys Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Ile Asp Ser Ser Ser Asn Ser Ala Ser Leu Thr Ile Ser Gly

65 70 75 80

Leu Thr Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Phe Asp Gly

85 90 95

Ser Ser Val Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 29

<211> 375

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1H2 VH

<400> 29

caggttcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaaaggag 300

cgtttctatg atagtagtgg ttattacgat gcttttgata tctggggcca agggacaatg 360

gtcaccgtctcttca 375

<210> 30

<211> 125

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1H2 VH

<400> 30

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Lys Glu Arg Phe Tyr Asp Ser Ser Gly Tyr Tyr Asp Ala Phe

100 105 110

Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120 125

<210> 31

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1H2 VL

<400> 31

cagtctgccc tgactcagcc tcgctcagtg tccggggtctc ctgggcagtc agtcaccatc 60

tcctgcactg gaaccagcaa tgatgttggt ggttataact atgtctcctg gtaccaacag 120

cacccaggca aagcccccaa actcatgatt tatgatgtca ctaagcggcc ctcaggggtc 180

cctgatcgct tctctggctc caagtctggc aacacggcct ccctgaccgt ctctggcctc 240

cagcctgagg atgaggctga ctattattgc gcctcttatg gaggcaggaa caatttgctt 300

tttggcggag ggactcaact gaccgtctta 330

<210> 32

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1H2 VL

<400> 32

Gln Ser Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Asn Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Pro Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Tyr Gly Gly Arg

85 90 95

Asn Asn Leu Leu Phe Gly Gly Gly Thr Gln Leu Thr Val Leu

100 105 110

<210> 33

<211> 354

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1E4 VH

<400> 33

caaatccagc tggtacaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc cggaggggga 300

gcagtggcgg acaatagtta ctggggccag ggaaccctgg tcaccgtctc ctca 354

<210> 34

<211> 118

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1E4 VH

<400> 34

Gln Ile Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr 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 Gly Gly Gly Ala Val Ala Asp Asn Ser Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 35

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1E4 VL

<400> 35

gacatccgga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctatgct gcatccagtt tacaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tggcacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtctacaa gattacaatt accctcgaac gttcggccaa 300

gggaccaagg tggaaatcaa a 321

<210> 36

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1E4 VL

<400> 36

Asp Ile Arg 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 Gly Ile Arg Asn Asp

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys 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 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 Leu Gln Asp Tyr Asn Tyr Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 37

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1A8 VH

<400> 37

caaatccagc tggtacaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacggcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagacggt 300

tcgtatagca gcagctggta ctcgtttgac tactggggcc agggaaccct ggtcaccgtc 360

tcctca 366

<210> 38

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1A8 VH

<400> 38

Gln Ile Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Asp Gly Ser Tyr Ser Ser Ser Trp Tyr Ser Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 39

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1A8 VL

<400> 39

cagtctgccc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc gatcaccatc 60

tcctgcactg gaaccagcag tgacgtcggt ggttataact atgtctcctg gtaccaacag 120

cacccaggca aagcccccaa actcatgatt tatgatgtca gtaatcggcc ctcaggggtt 180

tctaatcgct tctctggctc caagtctggc aacacggcct ccctgaccat ctctgggctc 240

caggctgagg acgaggctga ttattactgc tcctcatatg caggtgatat tagttatgta 300

ctgttcggcg gcgggaccaa gctgaccgtc cta 333

<210> 40

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1A8 VL

<400> 40

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Asp

85 90 95

Ile Ser Tyr Val Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 41

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1B11 VH

<400> 41

gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggagggtc cctgagactc 60

tcctgtgcag cctctggatt cacttttagt gactatgaca tgatctgggt ccgccaggct 120

ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180

gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gaaagagttc 300

tttggtgctt ttgatatctg gggccaaggg acaatggtca ccgtctcttc a 351

<210> 42

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1B11 VH

<400> 42

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

Asp Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala 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 Lys Glu Phe Phe Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Met

100 105 110

Val Thr Val Ser Ser

115

<210> 43

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1B11 VL

<400> 43

tcttctgagc tgactcagga ccctgctgtg tcggtggcct tgggacagac agtcacgatc 60

acatgccaag gagacagcct caattactat tatgcaaact ggttccagct gaagccagg 120

caggcccctg tacttgtcct ctttggtaaa aacaaccggc cctcagggat cccagaccga 180

ttctctggct cctactcggg aagcacagct tccttgacca tcactggggc tcaggcggaa 240

gatgacgctg actattactg taattcgcgg gacagcggtg gtaatccttg ggtgttcggc 300

ggagggacca agctgaccgt ccta 324

<210> 44

<211> 108

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1B11 VL

<400> 44

Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Thr Ile Thr Cys Gln Gly Asp Ser Leu Asn Tyr Tyr Tyr Ala

20 25 30

Asn Trp Phe Gln Leu Lys Pro Gly Gln Ala Pro Val Leu Val Leu Phe

35 40 45

Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Tyr Ser Gly Ser Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu

65 70 75 80

Asp Asp Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Gly Gly Asn Pro

85 90 95

Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 45

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1F3 VH

<400> 45

caaatccagc tggtacaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaagg atcatcccta tccttggtat agcagactac 180

gcacagaagt tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggaactga gtagcctggg atctgaggac acggccgtgt atttttgtgc gagagagggg 300

ggatccttta ggcactttga cttctggggc cagggaaccc tggtcaccgt ctcctca 357

<210> 46

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1F3 VH

<400> 46

Gln Ile Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asp Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Gly Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Glu Gly Gly Ser Phe Arg His Phe Asp Phe Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 47

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1F3 VL

<400> 47

cagcctgtgc tgactcagcc accctcagtc tctggggccc cagggcagag ggtcaccatc 60

tcctgcgctg ggagcgaccc caacatcggg acaggtcatg atgtgcactg gtaccagcaa 120

cttccaggaa cagcccccaa actcgtcatc tatggtaaca ccaatcggcc ctcaggggtc 180

cctgagcgat tcactgcctc caagtctggc acctcagcct ccctggccat cactgggctc 240

caggctgagg atgaggctga ttattactgc caggcctacg acaggagcct gcgtggttat 300

gtcttcggga ctgggaccaa ggtcaccgtc ctg 333

<210> 48

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1F3 VL

<400> 48

Gln Pro Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ala Gly Ser Asp Pro Asn Ile Gly Thr Gly

20 25 30

His Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

35 40 45

Val Ile Tyr Gly Asn Thr Asn Arg Pro Ser Gly Val Pro Glu Arg Phe

50 55 60

Thr Ala Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Tyr Asp Arg Ser

85 90 95

Leu Arg Gly Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 49

<211> 369

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1G5 VH

<400> 49

caaatccagc tggtacagtc tggtgctgaa gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaaga cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagaactaca 300

ggtgacgagt ggctacgatt ggctataaat gactactggg gccagggaac cctggtcacc 360

gtctcctca 369

<210> 50

<211> 123

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1G5 VH

<400> 50

Gln Ile 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 Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Thr Gly Asp Glu Trp Leu Arg Leu Ala Ile Asn Asp Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 51

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1G5 VL

<400> 51

gatattgtga tgacacagtc tcccctctcc ctgcccgtca cccctggaga gccggcctcc 60

atctcctgca ggtctagtct gcgcctcatg catcctaatg gactcaacta tttggattgg 120

tacctgcaga agccagggca gtctccacag ctcctaatct ttttgggttc tcagcgggcc 180

tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc 240

agcagagtgg aggctgagga tgttggcatt tattactgca tgcaagctct agaacctccg 300

tacacttttg gccaggggac caagctggag atcaaa 336

<210> 52

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1G5 VL

<400> 52

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Leu Arg Leu Met His Pro

20 25 30

Asn Gly Leu Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Phe Leu Gly Ser Gln Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Met Gln Ala

85 90 95

Leu Glu Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 53

<211> 372

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1C9 VH

<400> 53

caggtccagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagatccc 300

gggtatagca gtggctggaa agatgatgct tttgatatct ggggccaagg gacaatggtc 360

accgtctctt ca 372

<210> 54

<211> 124

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1C9 VH

<400> 54

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Asp Pro Gly Tyr Ser Ser Gly Trp Lys Asp Asp Ala Phe Asp

100 105 110

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 55

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1C9 VL

<400> 55

gaaattgtga tgacacagtc tccaggcacc ctgtctttgt ctccagggga tacagcctcc 60

ctctcctgca gggccagtca gactgttagc agcaactact tagcctggta ccaacagaaa 120

cctggccagg ctcccaggct cctcatctat gatacatcca acagggccgc tggcatcccg 180

gccaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag tagcctagag 240

cctgaagatt ttgcagtgta ttactgtcag cagtacggta gctcactctg gacgttcggc 300

caagggacca aggtggaaat caaa 324

<210> 56

<211> 108

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1C9 VL

<400> 56

Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Asp Thr Ala Ser Leu Ser Cys Arg Ala Ser Gln Thr Val Ser Ser Asn

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Asp Thr Ser Asn Arg Ala Ala Gly Ile Pro Ala Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Leu

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 57

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 11-A4 VH

<400> 57

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagcgggg 300

cagcagctgg tagccctttg gtactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 58

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 11-A4 VH

<400> 58

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Ala Gly Gln Gln Leu Val Ala Leu Trp Tyr Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 59

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 11-A4 VL

<400> 59

cagtctgccc tgactcagcc tccctccgcg tccgggtctc gtggacagtc agtctccatc 60

tcctgcagtg gaagtcgcag tgacattgga tattataact atgtctcctg gtatcaacaa 120

cacccaggca aagcccccaa actcatcatt tttgacgtca ataagcggcc ctcaggggtc 180

cctgatcgct tctctggctc caagtctggc aacacggcct ccctgaccgt ctctggcctc 240

cagcctgagg atgaggctga ctattattgc gcctcttatg gaggcaggaa caatttgctt 300

tttggcggag ggactcaact gaccgtctta 330

<210> 60

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 11-A4 VL

<400> 60

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Arg Gly Gln

1 5 10 15

Ser Val Ser Ile Ser Cys Ser Gly Ser Arg Ser Asp Ile Gly Tyr Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Ile Ile Phe Asp Val Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Pro Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Tyr Gly Gly Arg

85 90 95

Asn Asn Leu Leu Phe Gly Gly Gly Thr Gln Leu Thr Val Leu

100 105 110

<210> 61

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<223> 21-A1 VH

<400> 61

caggtgcaac tgcaggagtc gggcccagga ctggtggagc cttcggagac cctgtccctc 60

acctgcactg tctctggtgg ctccatcagt agtttctact ggagctggat ccggcagccc 120

ccagggaagg gactggagtg gattggctat atcaattaca gtgggagcac caactacaac 180

ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240

aagctgagct ctgtgaccgc cgcagacacg gctgtgtatt actgtgcgag acagatatta 300

tggttcgggg agttaaggtg gttcgacccc tggggccagg gaaccctggt caccgtctcc 360

tca 363

<210> 62

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-A1 VH

<400> 62

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Glu Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Phe

20 25 30

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Gln Ile Leu Trp Phe Gly Glu Leu Arg Trp Phe Asp Pro Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 63

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 21-A1 VL

<400> 63

cagtctgccc tgactcagcc tccctccgcg tccgggtctc ctggacagtc agtcaccatc 60

tcctgcactg gaaccagcag tgacattggt ggttataact atgtctcctg gtaccaactg 120

cgcccaggca aagcccccaa actcatgatt tatgacgtca ccaagcggcc ctcaggggtc 180

cctgatcgct tctctggctc caagtctggc aacacggcct ccctgaccgt ctctgggctc 240

caggctgagg atgaggctga ttattactgc agctcatatg caggcagcaa caatgtggta 300

ttcggcggag ggaccaagct gaccgtccta 330

<210> 64

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-A1 VL

<400> 64

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Leu Arg Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser

85 90 95

Asn Asn Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 65

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> 21-H12 VH

<400> 65

caagtccagc tggtacagtc tggggctgag gtgaagaagc ctggggcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaaatccc 300

tacggtttca actggttcga cccctggggc cagggaaccc tggtcaccgt ctcctca 357

<210> 66

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-H12 VH

<400> 66

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 Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Asn Pro Tyr Gly Phe Asn Trp Phe Asp Pro Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 67

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 21-H12 VL

<400> 67

aattttatgc tgactcagcc ccactctgtg tcggagtctc cggggaagac ggtaaccatc 60

tcctgcaccc gcagcagtgg cagcattgcc agcaactatg tgcagtggta ccagcagcgc 120

ccgggcagtg cccccaccac tgtgatctat gaggataacc aaagaccctc tggggtccct 180

gatcggttct ctggctccat cgacagctcc tccaactctg cctccctcac catctccgga 240

ctgaagactg aggacgaggc tgactactac tgtcagtctt atgatggctt caatcaggtg 300

ttcggcggag ggaccaagct gaccgtccta 330

<210> 68

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-H12 VL

<400> 68

Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Glu Ser Pro Gly Lys

1 5 10 15

Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ser Ile Ala Ser Asn

20 25 30

Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro Thr Thr Val

35 40 45

Ile Tyr Glu Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Ile Asp Ser Ser Ser Asn Ser Ala Ser Leu Thr Ile Ser Gly

65 70 75 80

Leu Lys Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Gly

85 90 95

Phe Asn Gln Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 69

<211> 372

<212> DNA

<213> Artificial sequence

<220>

<223> 7-D12 VH

<400> 69

caaatgcagc tggtacagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaaccggt 300

agtagtggtt atgtacgttg gagcaactgg ttcgacccct ggggccaggg aaccctggtc 360

accgtctcct ca 372

<210> 70

<211> 124

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 7-D12 VH

<400> 70

Gln Met Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr 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 Thr Gly Ser Ser Gly Tyr Val Arg Trp Ser Asn Trp Phe Asp

100 105 110

Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 71

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 7-D12 VL

<400> 71

gacatccaga tgacccagtc tccctccacc ctgtctgcat ttgtaggaga cagagtcacc 60

atcacttgcc gggccagtga gagtattagt aggtggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaaactcct aatctctaag acgtctaatt tagaaagcgg ggtcccgtca 180

aggttcagtg gcgctggatc tgggacagat ttcactctca ccattagcag tctgcaacct 240

gaggattttg caacttactt ctgtcaacag ggttccaaaa tgcctccgac tttcggcgga 300

gggaccaagg tggagatcaa g 321

<210> 72

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 7-D12 VL

<400> 72

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Phe Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Ile Ser Arg Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Ser Lys Thr Ser Asn Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ala 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 Phe Cys Gln Gln Gly Ser Lys Met Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 73

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 9-E3 VH

<400> 73

caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaggggcc 300

tacggtggta actccgcttt tgactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 74

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 9-E3 VH

<400> 74

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Gly Ala Tyr Gly Gly Asn Ser Ala Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 75

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> 9-E3 VL

<400> 75

cagtctgtgc tgacgcagcc gccctcagtg tctggggccc cagggcagag ggtcaccatc 60

tcctgcactg ggagcagctc caacatcggg gcaggttatg atgtacactg gtaccagcag 120

cttccaggaa cagcccccaa actcctcatg tacagtaatg atcagcggcc ctcaggggtc 180

actgagcgat tctctggctc caagtctggc acctcagcct ccctggccat cagtgggctc 240

cagtctgaag atgaggtga ttactactgc cagtcctatg acagaagcct gagaggttcg 300

gtcttcggcg gagggaccaa gctgaccgtc ctc 333

<210> 76

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 9-E3 VL

<400> 76

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly

20 25 30

Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu

35 40 45

Leu Met Tyr Ser Asn Asp Gln Arg Pro Ser Gly Val Thr Glu Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu

65 70 75 80

Gln Ser Glu Asp Glu Gly Asp Tyr Tyr Cys Gln Ser Tyr Asp Arg Ser

85 90 95

Leu Arg Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 77

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<223> 10-A6 VH

<400> 77

gaggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60

tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagattcc 300

atagcagcag ctggtactcc gttcgactac tggggccagg gaaccctggt caccgtctcc 360

tca 363

<210> 78

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 10-A6 VH

<400> 78

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 Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Ser Ile Ala Ala Ala Gly Thr Pro Phe Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 79

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 10-A6 VL

<400> 79

aattttatgc tgactcagcc ccactctgtg tcggagtctc cggggaagac ggtcaccatc 60

tcctgcaccc gcagcagtgg catcattgcc agcaaatatg tgcactggta ccagcagcgc 120

ccgggcagtg cccccaccac tgtgatctat gaggataacc aaagaccgtc tggggtccct 180

gatcgattct ctggctccat cgacaactcc tccaactctg cctccctcac catctctgga 240

ctgcagactg aggacgaggc tgactactac tgtcagtctc atgacggcat caatcaggtt 300

ttcggcggag ggaccaaggt caccgtccta 330

<210> 80

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 10-A6 VL

<400> 80

Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Glu Ser Pro Gly Lys

1 5 10 15

Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ile Ile Ala Ser Lys

20 25 30

Tyr Val His Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro Thr Thr Val

35 40 45

Ile Tyr Glu Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Ile Asp Asn Ser Ser Asn Ser Ala Ser Leu Thr Ile Ser Gly

65 70 75 80

Leu Gln Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser His Asp Gly

85 90 95

Ile Asn Gln Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 81

<211> 372

<212> DNA

<213> Artificial sequence

<220>

<223> 12-A4 VH

<400> 81

gaggtgcagc tggtggagtc ccggggaggc ttggtacagc cggggggggtc cctgagactc 60

tcctgtgtaa cttctggatt cagctttaac aactatgcca tgaactgggt ccgccaggct 120

ccggggaagg ggctggagtg ggtctcagct gttagtggta gtggtggtac cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgttt 240

gtgcagatgg acagcctgag agctgaggac acggctgtgt attactgtgc gaaaggactt 300

ttccctacga tttttggagt aggagcaatg tttgactact ggggccaggg aaccctggtc 360

accgtctcct ca 372

<210> 82

<211> 124

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 12-A4 VH

<400> 82

Glu Val Gln Leu Val Glu Ser Arg Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Thr Ser Gly Phe Ser Phe Asn Asn Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Val Ser Gly Ser Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Val Gln Met Asp Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Gly Leu Phe Pro Thr Ile Phe Gly Val Gly Ala Met Phe Asp

100 105 110

Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 83

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> 12-A4 VL

<400> 83

tcttctgagc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60

tcttgttctg gaagcagctc caacatcgga agtaatgctg ttaactggta tcagcagctc 120

cccaggaacgg cccccaaact cctcatctat gataataatc accggccctc aggggtccct 180

gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240

tctgaggatg aggctgatta ttattgtgca gcatgggatg acaccattcc tggtgtgcta 300

ttcgccggag ggaccaagct gaccgtccta 330

<210> 84

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 12-A4 VL

<400> 84

Ser Ser Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn

20 25 30

Ala Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn His Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Thr Ile

85 90 95

Pro Gly Val Leu Phe Ala Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 85

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> 14-G10 VH

<400> 85

gaagtgcagc tggtggagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaggtgtt 300

tcttactact acggtatgga cgtctggggc caagggacca cggtcaccgt ctcctca 357

<210> 86

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 14-G10 VH

<400> 86

Glu Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Gly Val Ser Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly

100 105 110

Thr Thr Val Thr Val Ser Ser

115

<210> 87

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> 14-G10 VL

<400> 87

caggctgtgc tgactcagcc accctcggtg tccgtgtccc caggacagac agccatcatc 60

tcctgttctg gacataaatt gggtgataag tatgtttcct ggtatcaaca gcagccaggc 120

cagtcccctg tgctggtcct ctttcaggat accaagcggc cctcagggat ccctgagcga 180

ttctctggct ccaactctgg gaacacagcc actctgacca tcagcgcgac ccaggctgcg 240

gatgaggctg actattactg tcaggcgggg gacaccaagt ctgtgatctt cggcggcggg 300

accaagctga ccgtccta 318

<210> 88

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 14-G10 VL

<400> 88

Gln Ala Val Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln

1 5 10 15

Thr Ala Ile Ile Ser Cys Ser Gly His Lys Leu Gly Asp Lys Tyr Val

20 25 30

Ser Trp Tyr Gln Gln Gln Pro Gly Gln Ser Pro Val Leu Val Leu Phe

35 40 45

Gln Asp Thr Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Ala Thr Gln Ala Ala

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Gly Asp Thr Lys Ser Val Ile

85 90 95

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 89

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 22-A6 VH

<400> 89

caggttcagg tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcggcaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagaggatac 300

agctatggtt caggacacct tgactactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 90

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 22-A6 VH

<400> 90

Gln Val Gln Val Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Gly Tyr Ser Tyr Gly Ser Gly His Leu Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 91

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 22-A6 VL

<400> 91

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc aggcgagtca ggacattagc aactatttaa attggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctacgat gcatccaatt tggaaacagg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcgctctca ccatcagcag tctccaacct 240

gaagatttg caacttatta ctgtctacag cataatagtt accctcggac ttttggccag 300

gggaccaagc tggagatcaa a 321

<210> 92

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 22-A6 VL

<400> 92

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 Gln 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 Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Ala Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 93

<211> 360

<212> DNA

<213> Artificial sequence

<220>

<223> 35-B1 VH

<400> 93

gaggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggata caccttcacc ggctactata tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atgaacccta acagtggtga cacagcctat 180

acacagaact tccagggcag agtcaccatg accaggaacc cctccataag cacagcctac 240

atggagctga gcaacctgag atctgaggac acggccgtgt attactgtgc gagaggccgg 300

gggttcgcgg agaagcccct tgggtactgg ggccagggaa ccctggtcac cgtctcctca 360

<210> 94

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 35-B1 VH

<400> 94

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 Gly Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Met Asn Pro Asn Ser Gly Asp Thr Ala Tyr Thr Gln Asn Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asn Pro Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Asn Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Arg Gly Phe Ala Glu Lys Pro Leu Gly Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 95

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<223> 35-B1 VL

<400> 95

gatattgtga tgactcagtc tccagactcc ctggctgtgt ctctgggcgg gagggccacc 60

atcaactgca agtccagcca gagtatttta tccagctcca ataataagaa ctatttagct 120

tggtaccagc agaaaccagg tcagcctcct aagctgctca tttactgggc atctacccgg 180

gaatccgggg tccctgaccg gttcagcggc agcgggtctg ggacagattt cactctcacc 240

atcagcagcc tgcaggctga agatgtggca gtttattact gtcagcaata ttatagtact 300

cctccgacat tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 96

<211> 113

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 35-B1 VL

<400> 96

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gly Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Ile Leu Ser Ser

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys 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 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 97

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1F4 VH

<400> 97

gaggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180

gcacagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagggcccct 300

cgagggcagt ggctggttca ctactttgac tactggggcc agggaaccct ggtcaccgtc 360

tcctca 366

<210> 98

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1F4 VH

<400> 98

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr 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 Ala Pro Arg Gly Gln Trp Leu Val His Tyr Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 99

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 3-1F4 VL

<400> 99

gaaattgtgt tgacgcagtc tccagccacc ctctctctgt ctccagggga aagagccacc 60

ctctcctgct gggccagtca ggatgttagc aactacttag cctggtacca acagaagcct 120

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 180

aggttcagcg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240

gaagattttg cagtgtatta ctgtcagcaa cgtagcaact ggcctctcac tttcggcggc 300

gggaccaagg tggagctcaa a 321

<210> 100

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 3-1F4 VL

<400> 100

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 Trp Ala Ser Gln Asp Val Ser Asn Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Leu Lys

100 105

<210> 101

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B3 VH

<400> 101

caggttcagc tggtgcagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180

gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240

atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagagtcc 300

tactcgtccg caggtattga ctactggggc cagggaaccc tggtcaccgt ctcctca 357

<210> 102

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B3 VH

<400> 102

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 Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Ser Tyr Ser Ser Ala Gly Ile Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 103

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<223> 4-1B3 VL

<400> 103

gatattgtga tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc 60

atctcctgca ggtctagtca gaccctcctg catagtaatg gattcaacta tttggattgg 120

tacctgcaga agccagggca gtctccacaa ctcctgatgt atttgggctc tagccgggcc 180

tccggggtcc ctgacaggtt cagtggcagt ggatcgggca cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tgttggggtt tattactgca tgcaaactct acaaactcct 300

ccggctttcg gcggagggac caaggtggag atcaaa 336

<210> 104

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 4-1B3 VL

<400> 104

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Thr Leu Leu His Ser

20 25 30

Asn Gly Phe Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Met Tyr Leu Gly Ser Ser Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Thr

85 90 95

Leu Gln Thr Pro Pro Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 105

<211> 354

<212> DNA

<213> Artificial sequence

<220>

<223> 21-G1 VH

<400> 105

caggtccagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgacgatt 60

tcctgcgagg cgtctggata caacttcatc agctactata tacactgggt gcgacaggcc 120

cctggacaag gccttgagtg gatgggattc gtcgtcccta gtggtggtgc cgcaggctac 180

acacagaagt tccagggcag actcaccgtg accagggaca cgtccacgag cacagtctac 240

atggacctga acagcctgac atctgacgac acggccgtgt attactgtgt gcgagaaatg 300

agtggtggct ggtttgattt ctggggccag ggaaccctgg tcaccgtctc ctcg 354

<210> 106

<211> 118

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-G1 VH

<400> 106

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Thr Ile Ser Cys Glu Ala Ser Gly Tyr Asn Phe Ile Ser Tyr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Phe Val Val Pro Ser Gly Gly Ala Ala Gly Tyr Thr Gln Lys Phe

50 55 60

Gln Gly Arg Leu Thr Val Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Asp Leu Asn Ser Leu Thr Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Arg Glu Met Ser Gly Gly Trp Phe Asp Phe Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 107

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> 21-G1 VL

<400> 107

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gagcattagc agctatttaa attggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180

aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttacta ctgtcaacag agttacagta ccccgatcac cttcggccaa 300

ggggacacgac tggagattaa a 321

<210> 108

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> 21-G1 VL

<400> 108

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 Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys 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 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 Ser Tyr Ser Thr Pro Ile

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

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Claims (40)

1. An antigen-binding polypeptide that binds to a human PD-L1 epitope, comprising a heavy chain variable domain and a light chain variable domain, wherein their respective sequences consist essentially of pairs of sequences selected from the group consisting of: (a) SEQ ID NO: 18 and SEQ ID NO: 20; (b) SEQ ID NO: 42 and SEQ ID NO: 44; (c) SEQ ID NO: 34 and SEQ ID NO: 36; (d) SEQ ID NO: 22 and SEQ ID NO: 24; (e) SEQ ID NO: 2 and SEQ ID NO: 4; (f) SEQ ID NO: 62 and SEQ ID NO: 64; (g) SEQ ID NO: 82 or SEQ ID NO: 84; (h) SEQ ID NO: 70 and SEQ ID NO: 72; (i) SEQ ID NO: 50 and SEQ ID NO: 52; (j) SEQ ID NO: 102, and SEQ ID NO: 104; (k) SEQ ID NO: 30 and SEQ ID NO: 32; (l) SEQ ID NO: 6 and SEQ ID NO: 8; (m) SEQ ID NO: 10 and SEQ ID NO: 12; (n) SEQ ID NO: 14 and SEQ ID NO: 16; (o) SEQ ID NO: 26 and SEQ ID NO: 28; (p) SEQ ID NO: 38 and SEQ ID NO: 40; (q) SEQ ID NO: 46 and SEQ ID NO: 48; (r) SEQ ID NO: 54 and SEQ ID NO: 56; (s) SEQ ID NO: 58 and SEQ ID NO: 60; (t) SEQ ID NO: 66 and SEQ ID NO: 68; (u) SEQ ID NO: 74 and SEQ ID NO: 76; (v) SEQ ID NO: 78 and SEQ ID NO: 80; (w) SEQ ID NO: 86 and SEQ ID NO: 88; (x) SEQ ID NO: 90 and SEQ ID NO: 92; (y) SEQ ID NO: 94 and SEQ ID NO: 96; (z) SEQ ID NO: 98 and SEQ ID NO: 100; And (aa) SEQ ID NO: 106 and SEQ ID NO: 108. 2. The antigen-binding polypeptide according to claim 1, characterized in that the heavy chain variable domain and the light chain variable domain of said polypeptide have amino acid sequences consisting of SEQ ID NO: 18 and SEQ ID NO: 20. 3. The antigen-binding polypeptide of claim 1, wherein the heavy chain variable domain and the light chain variable domain have amino acid sequences consisting of SEQ ID NO: 42 and SEQ ID NO: 44. 4. The antigen-binding polypeptide according to claim 1, characterized in that the heavy chain variable domain and the light chain variable domain have the amino acid sequences of SEQ ID NO: 34 and SEQ ID NO: 36. 5. Antigen-binding polypeptide according to claims. 1-4, characterized in that said polypeptide is a fully human antibody or a humanized antibody. 6. Antigen-binding polypeptide according to claim 5, characterized in that said antibody contains a human constant region having ADCC activity and/or CDC activity. 7. A nucleic acid molecule encoding an antigen-binding polypeptide according to claims. 1-4, wherein said nucleic acid molecule is a DNA molecule or an RNA molecule. 8. The nucleic acid molecule according to claim 7, wherein said nucleic acid molecule contains DNA sequences encoding a heavy chain variable domain and a light chain variable domain, said DNA sequences consisting essentially of pairs of sequences selected from the group consisting of : (a) SEQ ID NO: 17 and SEQ ID NO: 19; (b) SEQ ID NO:33 and SEQ ID NO:35; and (c) SEQ ID NO:41 and SEQ ID NO:43. 9. A pharmaceutical composition for use in inhibiting the biological activity of PD-L1, containing an effective amount of an antigen-binding polypeptide according to any one of paragraphs. 1-6 and a pharmaceutically acceptable excipient, pharmaceutically acceptable carrier or diluent. 10. A method of therapeutically treating a subject in need thereof of a condition, said method comprising administering to said subject a therapeutically effective amount of an antigen-binding polypeptide according to any one of claims. 1-6, characterized in that the condition is a cancer containing at least a portion of tumor cells expressing a detectable amount of PD-L1. 11. The method of claim 10, further comprising administration in combination with (a) antibodies targeting other immunosuppressive pathways; (b) chemotherapy or radiation therapy; (c) other mechanisms of blocking immunosuppressive pathways; or (d) other immunotherapy agents. 12. Method according to any one of paragraphs. 10, 11, characterized in that the specified condition is a mammalian cancer selected from the group consisting of: ovarian cancer, colon cancer, breast cancer, lung cancer, myelomas, tumors of the central nervous system of neuroblastic origin, monocytic leukemias, B-cell leukemias , T-cell leukemias, B-cell lymphomas, T-cell lymphomas, mast cell-derived tumors, melanoma, bladder cancer, gastric cancer, liver cancer, urothelial carcinoma, cutaneous carcinoma, kidney cancer, head and neck cancer, cancer pancreas and combinations of the above.