Anti-CD73 antibodies and uses thereof

Abstract

To provide anti-CD73 antibodies and antigen-binding fragments thereof.SOLUTION: The present invention provides anti-CD73 antibodies that include HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 with specific amino acid sequences and specifically bind to CD73, preferably human CD73, and antigen-binding fragments thereof. Also provide are nucleic acids encoding the antibodies, compositions comprising the antibodies, and methods of producing the antibodies and using the antibodies for treating or preventing diseases such as cancer.SELECTED DRAWING: Figure 1A-1B

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 62 / 640,955, filed March 9, 2018; U.S. Provisional Application No. 62 / 721,044, filed August 22, 2018; and U.S. Provisional Application No. 62 / 786,598, filed December 31, 2018, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] FIELD OF THEINVENTION The present invention relates to monoclonal anti-CD73 antibodies, nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. Methods of making the antibodies and methods of using the antibodies to treat diseases associated with CD73, such as cancer and inflammatory diseases, and / or associated complications, are also provided.

[0003] Reference to an Electronically Submitted Sequence Listing This application contains a Sequence Listing submitted electronically via EFS-Web as an ASCII formatted Sequence Listing having a size of 114 kb, with a filename of "689204.10WO Sequence Listing" and a creation date of February 24, 2019. The Sequence Listing submitted via EFS-Web is a part of this specification and is incorporated by reference in its entirety. [Background technology]

[0004] CD73, also known as ecto-5'-nucleotidase (ecto-5'-NT or EC 3.1.3.5), is a cell surface phosphatase that catalyzes the dephosphorylation of extracellular AMP to generate adenosine. CD73 is a glycophosphatidylinositol (GPI)-anchored protein that can be released to generate a catalytically active form (Arias et al., J Cell Biol. 1997; 136(2):421-31). Physiologically, CD73 is induced by hypoxia and controls inflammation at wound sites (Bours et al., Pharmacol Ther 2006; 112:358-404). Pathologically, CD73 is often found overexpressed in regulatory T (Treg) cells and tumor cells (Paul et al., Trends Immunol 2012; 33:231-237). Increased CD73 activity leads to the accumulation of adenosine in the tumor microenvironment (TME).

[0005] Accumulating evidence indicates that CD73 activity at tumor sites is one of the key factors shaping the tumor-promoting TME that plays a critical role in tumor growth and survival (Whiteside, Expert Rev. Anticancer Ther. 2017; 17(6):527-35). Extracellular ATP-adenosine homeostasis is determined by the activity of CD39, which converts ATP to AMP, and CD73, which uses AMP to generate adenosine. By binding to the adenosine receptors A2a and A2b, adenosine suppresses both innate and adaptive immunity by modulating many immune cells, such as macrophages (Csoka et al., FASEB J 2012; 26:376-386), dendritic cells (Panther et al., Blood 2003; 101:3985-3990), natural killer cells (Hausler et al., Cancer Immunol Immunother 2011; 60:1405-1418), and effector T cells (Hoskin et al., Int J Oncol 2008; 32:527-535). It has therefore been hypothesized that adenosine-induced immune suppression may be alleviated by inhibiting the enzymatic activity of CD73 in the TME. Indeed, in vivo animal studies (Jin et al., Cancer Res 2010; 70:2245-2255; Stagg et al., Cancer Res 2011; 71:2890-2900) have shown that inhibiting the enzymatic activity of CD73 suppresses tumor formation and growth, suggesting that CD73 is a promising target for cancer therapy. Monoclonal antibodies that inhibit the enzymatic activity of CD73 and / or reduce CD73 content at the cell surface (i.e., by inducing internalization of CD73) may be effective in treating cancer, either alone as monotherapy or in combination with other cancer immunotherapeutics and / or other types of anti-cancer therapies. Summary of the Invention

[0006] In one general aspect, the invention relates to an isolated monoclonal antibody, or antigen-binding fragment thereof, that binds to CD73.

[0007] (1) SEQ ID NOs: 89, 90, 91, 149, 150, and 151, respectively; (2) SEQ ID NOs: 41, 42, 43, 101, 102, and 103, respectively; (3) SEQ ID NOs: 44, 45, 46, 104, 105, and 106, respectively; (4) SEQ ID NOs: 47, 48, 49, 107, 108, and 109, respectively; (5) SEQ ID NOs: 50, 51, 52, 110, 111, and 112, respectively; (6) SEQ ID NOs: 53, 54, 55, 113, 114, and 115, respectively; (7) SEQ ID NOs: 56, 57, 58, 116, 117, and 118, respectively; (8) SEQ ID NOs: 59, 60, 61, 119, 120, and 121, respectively; (9) SEQ ID NOs: 62, 63, 64, 122, 123, and 124, respectively; (10) SEQ ID NOs: 65, 66, 67, 125, 126, and 127, respectively; (11) SEQ ID NOs: 68, 69, 70, 128, 129, and 130, respectively; (12) SEQ ID NOs: 71, 72, 73, 131, 132, and 133, respectively; (13) SEQ ID NOs: 74, 75, 76, 134, 135, and 136, respectively; (14) SEQ ID NOs: 77, 78, 79, 137, 138, and 139, respectively; (15) SEQ ID NOs: 80, 81, 82, 140, 141, and 142, respectively; (16) SEQ ID NOs: 83, 84, 85, 143, 144, and 145, respectively; (17) SEQ ID NOs: 86, 87, 88, 146, 147, and 148, respectively; (18) SEQ ID NOs: 92, 93, 94, 152, 153, and 154, respectively; (19) SEQ ID NOs: 95, 96, 97, 155, 156, and 157, respectively; or (20) SEQ ID NOs: 98, 99, 100, 158, 159, and 160, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0008] (1) SEQ ID NOs: 209, 210, 211, 269, 270, and 271, respectively; (2) SEQ ID NOs: 161, 162, 163, 221, 222, and 223, respectively; (3) SEQ ID NOs: 164, 165, 166, 224, 225, and 226, respectively; (4) SEQ ID NOs: 167, 168, 169, 227, 228, and 229, respectively; (5) SEQ ID NOs: 170, 171, 172, 230, 231, and 232, respectively; (6) SEQ ID NOs: 173, 174, 175, 233, 234, and 235, respectively; (7) SEQ ID NOs: 176, 177, 178, 236, 237, and 238, respectively; (8) SEQ ID NOs: 179, 180, 181, 239, 240, and 241, respectively; (9) SEQ ID NOs: 182, 183, 184, 242, 243, and 244, respectively; (10) SEQ ID NOs: 185, 186, 187, 245, 246, and 247, respectively; (11) SEQ ID NOs: 188, 189, 190, 248, 249, and 250, respectively; (12) SEQ ID NOs: 191, 192, 193, 251, 252, and 253, respectively; (13) SEQ ID NOs: 194, 195, 196, 254, 255, and 256, respectively; (14) SEQ ID NOs: 197, 198, 199, 257, 258, and 259, respectively; (15) SEQ ID NOs: 200, 201, 202, 260, 261, and 262, respectively; (16) SEQ ID NOs: 203, 204, 205, 263, 264, and 265, respectively; (17) SEQ ID NOs: 206, 207, 208, 266, 267, and 268, respectively; (18) SEQ ID NOs: 212, 213, 214, 272, 273, and 274, respectively; (19) SEQ ID NOs: 215, 216, 217, 275, 276, and 277, respectively; or (20) SEQ ID NOs: 218, 219, 220, 278, 279, and 280, respectively; and a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0009] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37, or 39, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38, or 40.

[0010] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 33, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 34; (b) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2; (c) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4; (d) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6; (e) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8; (f) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10; (g) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12; (h) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; (i) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (j) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 17, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 18; (k) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 20; (l) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22; (m) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:23, and a light chain variable region having the polypeptide sequence of SEQ ID NO:24; (n) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26; (o) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28; (p) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30; (q) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 31, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 32; (r) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 35, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 36; (s) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 37, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 38; or (t) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 39, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40. Includes.

[0011] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof inhibits the enzymatic activity of soluble and / or cell surface CD73.

[0012] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof prevents dimerization of CD73.

[0013] In certain embodiments, the isolated monoclonal antibody, or antigen-binding fragment thereof, induces internalization of CD73.

[0014] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is chimeric.

[0015] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is human or humanized.

[0016] In certain embodiments, the humanized monoclonal antibody or antigen-binding fragment thereof has the following structure: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 282, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 287, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 288, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 289, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 295, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 296, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 297, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; or (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 298, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299. Includes.

[0017] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is capable of activating T cells.

[0018] Also provided is an isolated nucleic acid encoding the monoclonal antibodies, or antigen-binding fragments thereof, of the invention disclosed herein.

[0019] Also provided is a vector comprising an isolated nucleic acid encoding a monoclonal antibody, or antigen-binding fragment thereof, of the invention.

[0020] A host cell comprising a vector containing an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof of the invention is also provided.

[0021] In certain embodiments, a pharmaceutical composition is provided comprising an isolated monoclonal antibody or antigen-binding fragment thereof of the invention and a pharma- ceutically acceptable carrier.

[0022] Also provided is a method of inhibiting the nucleotidase activity of CD73 in a subject in need thereof, comprising the step of administering to the subject a pharmaceutical composition of the invention.

[0023] Also provided is a method of preventing dimerization of CD73 in a subject in need thereof, comprising the step of administering to the subject a pharmaceutical composition of the invention.

[0024] Also provided is a method of inducing internalization of CD73 in a subject in need thereof, comprising the step of administering to the subject a pharmaceutical composition of the invention.

[0025] Also provided is a method of treating cancer in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the present invention. The cancer may be any liquid or solid cancer, including, but not limited to, lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML), and other liquid tumors.

[0026] Also provided is a method for producing a monoclonal antibody or antigen-binding fragment thereof of the invention, comprising culturing a cell containing nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof under conditions to produce the monoclonal antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the cell or culture.

[0027] Also provided is a method for producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof of the invention, the method comprising combining the monoclonal antibody or antigen-binding fragment thereof with a pharma- ceutically acceptable carrier to obtain the pharmaceutical composition.

[0028] Also provided is a method of determining the level of CD73 in a subject, the method comprising the steps of: (a) obtaining a sample from the subject; (b) contacting the sample with an antibody or antigen-binding fragment thereof of the present invention; and (c) determining the level of CD73 in the subject. In certain embodiments, the sample is a tissue sample. The tissue sample may be, for example, a cancer tissue sample. In certain embodiments, the sample is a blood sample.

[0029] Also provided is a method for determining the ecto-5'-nucleotidase activity of CD73 in a subject, where the enzymatic activity can be completely inhibited by a monoclonal antibody or antigen-binding fragment thereof of the present invention. The method comprises the steps of (a) obtaining a sample from a subject; (b) contacting the sample with an antibody or antigen-binding fragment thereof of the present invention; and (c) determining the ecto-5'-nucleotidase activity of CD73 in the subject. In certain embodiments, the sample is a tissue sample. The tissue sample may be, for example, a cancer tissue sample. In certain embodiments, the sample is a blood sample.

[0030] The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the accompanying drawings. It should be understood, however, that the present application is not limited to the precise embodiments shown in the drawings. [Brief description of the drawings]

[0031] [Figure 1A-1B] 1A and 1B show the inhibition of nucleotidase activity of human CD73 immobilized on a plate by chimeric anti-CD73 mAb. No antibody is the enzyme reaction in the absence of added antibody; AMPCP (adenosine 5'(α,β-methylene)diphosphate) was used as a control for inhibition of enzyme activity. [Figure 2A-2B] 2A-2E show the binding of chimeric anti-CD73 mAbs to human CD73 in an ELISA assay. [Fig. 2C-2D] This is a continuation of Figures 2A-2B. [Figure 2E] This is a continuation of Figures 2C-2D. [Diagram 3] Figure 3 shows the inhibition of nucleotidase activity of soluble human CD73 by chimeric mAbs 60A9A / IgG4 and 39G8A / IgG4. No mAb, enzymatic reaction where no antibody was added; No CD73, negative control reaction where neither antibody nor CD73 was added. [Figure 4A-4B]Figures 4A-4D show binding to human CD73 and inhibition of nucleotidase activity of human CD73 by humanized mAb 60A9-H5L4 / IgG4. Figure 4A shows binding of humanized mAb 60A9-H5L4 / IgG4 to human CD73 in an ELISA assay; chimeric mAb 60A9A / IgG4 was used as a control. Figure 4B shows inhibition of nucleotidase activity of soluble human CD73 by humanized mAb 60A9-H5L4 / IgG4. No mAb, enzymatic reaction in the absence of added mAb; No CD73, enzymatic reaction in the absence of added CD73. Figure 4C shows inhibition of nucleotidase activity of human CD73 expressed on the surface of A375 cells by humanized mAb 60A9-H5L4 / IgG4; chimeric mAb 60A9A / IgG4 was used as a control. No mAb, enzyme reaction without mAb; no AMP, enzyme reaction without AMP; no cells, enzyme reaction without cells. Figure 4D shows the inhibition of nucleotidase activity of human CD73 expressed on the surface of A375 cells by humanized mAbs 60A9-H3L5 / IgG4, 60A9-H5L5 / IgG4, 60A9-H6L5 / IgG4, 60A9-H7L5 / IgG4, 60A9-H8L5 / IgG4 and 60A9-H9L5 / IgG4; chimeric mAb 60A9A / IgG4 was used as a control. No mAb, enzyme reaction without mAb; no AMP, enzyme reaction without AMP; no cells, enzyme reaction without cells. [Fig. 4C-4D] Continuation of Figures 4A-4B. [Diagram 5] Figure 5 shows inhibition of CD73 nucleotidase activity in patient serum samples by humanized anti-CD73 mAb 60A9-H5L4 / IgG4. No mAb is the enzyme reaction in the absence of added antibody; assay background is the enzyme reaction in the presence of 100 μM AMP and 100 μM ATP; cps is counts / second. [Figure 6] FIG. 6 shows activation of T cell proliferation by humanized anti-CD73 mAb 60A9-H5L4 / IgG4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Various publications, documents, and patents are cited or described in the Background and throughout this specification, and each of these references is incorporated herein by reference in its entirety. The discussion of documents, acts, materials, devices, articles and the like which has been included in this specification is for the purpose of providing a context for the present invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any invention disclosed or claimed.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Otherwise, certain terms used herein have the meanings set forth herein.

[0034] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0035] Unless otherwise stated, any numerical value, e.g., concentration or concentration range described herein, should be understood to be modified in all instances by the term "about." Thus, numerical values ​​typically include ±10% of the stated value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, the use of numerical ranges expressly includes all possible subranges, all individual numerical values ​​within such ranges, including integers within such ranges and fractions of such values, unless the context clearly indicates otherwise.

[0036] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0037] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," or "containing," or any other variations thereof, refer to the inclusion of a stated integer or group of integers, but not to the exclusion of any other integer or group of integers, and are intended to be non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or device that includes a list of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or device. Further, unless expressly stated to the contrary, "or" refers to an inclusive or and not an exclusive or. For example, conditions A or B may satisfy any one of the following: A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); and both A and B are true (or exist).

[0038] As used herein, the connective term "and / or" between multiple listed elements is understood to encompass both individual and combined options. For example, when two elements are connected by "and / or", the first option refers to the applicability of the first element without the second element. The second option refers to the applicability of the second element without the first element. The third option refers to the applicability of the first and second elements together. Any one of these options is understood to be within the meaning and thus meets the requirements of the term "and / or" as used herein. The simultaneous application of one or more of these options is also understood to be within the meaning and thus meets the requirements of the term "and / or".

[0039] As used herein, the term "consists of," or variations thereof, such as "consist of" or "consisting of," as used throughout the specification and claims, indicates the inclusion of any stated integer or group of integers, but does not allow additional integers or groups of integers to be added to a particular method, structure, or composition.

[0040] As used herein, the term "consists essentially of," or variations thereof, such as "consist essentially of" or "consisting essentially of," as used throughout the specification and claims, indicates the inclusion of any recited integer or group of integers, and the optional inclusion of any recited integer or group of integers that does not materially alter the basic or novel characteristics of a particular method, structure, or composition. See MPEP § 2111.03.

[0041] As used herein, "subject" refers to any animal, preferably a mammal, most preferably a human. The term "mammal" as used herein encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more preferably humans.

[0042] The words "right", "left", "lower" and "top" designate directions in the drawings to which reference is made.

[0043] It should also be understood that the terms "about," "approximately," "generally," "substantially," and the like, used herein when referring to dimensions or features of preferred inventive components, do not mean that the described dimensions / features are precise boundaries or parameters, but do not exclude minor variations thereof that are functionally the same or similar, as would be understood by one of ordinary skill in the art. At a minimum, such references involving numerical parameters include variations that do not alter at least the least significant digit when using mathematical and industrial principles accepted in the art (e.g., rounding errors, measurement errors or other systematic errors, manufacturing tolerances, and the like).

[0044] The terms "identical" or percent "identity," in the context of two or more nucleic acid or polypeptide sequences (e.g., anti-CD73 antibodies and the polynucleotides which encode them, CD73 polypeptides and the CD73 polynucleotides which encode them), refer to two or more sequences or subsequences that are the same, or have a particular percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence as determined using one of the following sequence comparison algorithms or by visual inspection.

[0045] In sequence comparison, one sequence usually serves as the reference sequence with which test sequence is compared.When using sequence comparison algorithm, test sequence and reference sequence are input into computer, subsequence coordinates are designated as necessary, and sequence algorithm program parameters are designated.Then, sequence comparison algorithm calculates the percent sequence identity for test sequence compared to reference sequence based on designated program parameters.

[0046] Optimal sequence alignment for comparison can be determined, for example, by the local homology algorithm of Smith and Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443 (1970), by the similarity search method of Pearson and Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (see generally Current Protocols in Molecular Biology, FM Ausubel et al., eds., Current Protocols, Greene Publishing Associates, Inc. and John Wiley & Sons, This may be done through a joint venture between Ausubel and Samsung Electronics, Inc. (see 1995 Addendum) (Ausubel).

[0047] Examples of suitable algorithms for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms described in Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm first involves identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that either match or meet a certain positive threshold score T when aligned with words of the same length in database sequences. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds to initiate searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased.

[0048] Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. Extension of the word hits in each direction is halted when the cumulative alignment score falls by an amount X from its maximum achieved value; when the cumulative score falls below 0 due to the accumulation of one or more negatively scoring residue alignments; or when the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)).

[0049] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul, Proc. Nat'l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the minimum sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered to be similar to a reference sequence if the minimum sum probability in the comparison between the test nucleic acid and the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0050] A further indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, when the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions.

[0051] As used herein, the terms "inhibit", "inhibiting" and "inhibition" refer to reducing an activity, response, condition, disease or other biological parameter. This can include, but is not limited to, the complete elimination of the activity, response, condition, or disease. This can also include, for example, a 10% reduction in the activity, response, condition, or disease compared to native or control levels. Thus, the reduction can be a 10, 20, 30, 40, 50, 60, 70, 80, 90, 100% reduction, or any amount in between, compared to native or control levels. As a non-limiting example, an antibody of the invention can inhibit the nucleotidase activity of CD73 protein. The activity of CD73 protein can be reduced or eliminated compared to the activity of native CD73 protein. As another non-limiting example, an antibody of the invention can inhibit or impede (prevent or prevent) dimerization of CD73 protein in a subject. Dimerization of CD73 can be reduced or eliminated compared to native CD73 dimerization.

[0052] antibody The present invention relates generally to isolated anti-CD73 antibodies, nucleic acids and expression vectors encoding these antibodies, recombinant cells containing the vectors, and compositions comprising these antibodies. The present invention also relates generally to methods of making these antibodies, and methods of using these antibodies to treat diseases, including cancer. The antibodies of the present invention have one or more desirable functional properties, including, but not limited to, high affinity binding to CD73, high specificity for CD73, the ability to inhibit the nucleotidase activity of CD73, the ability to prevent dimerization of CD73, the ability to induce internalization of CD73 into cells (resulting in a reduction in the cell surface content of CD73), and the ability to inhibit tumor growth in subjects and animal models in need thereof when administered alone or in combination with other anti-cancer therapies.

[0053] In a general aspect, the present invention relates to an isolated monoclonal antibody, or antigen-binding fragment thereof, that specifically binds to CD73.

[0054] As used herein, the term "antibody" is used in a broad sense and includes immunoglobulin or antibody molecules, including monoclonal or polyclonal, human, humanized, composite and chimeric antibodies and antibody fragments. In general, antibodies are proteins or peptide chains that exhibit binding specificity to a specific antigen. Antibody structure is well known. Immunoglobulins can be assigned to five major classes (i.e., IgA, IgD, IgE, IgG and IgM) depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3 and IgG4. Thus, the antibodies of the present invention may be of any of the five major classes or corresponding subclasses. Preferably, the antibodies of the present invention are IgG1, IgG2, IgG3 or IgG4. Antibody light chains of vertebrate species can be assigned to one of two clearly distinct types, namely kappa and lambda, based on the amino acid sequences of their constant domains. Thus, the antibodies of the present invention may contain a kappa or lambda light chain constant domain. According to certain embodiments, the antibodies of the invention comprise heavy and / or light chain constant regions derived from a rat or human antibody. In addition to the heavy and light chain constant domains, the antibodies comprise an antigen-binding region composed of a light chain variable region and a heavy chain variable region, each of which comprises three domains (i.e., complementarity determining regions 1-3; CDR1, CDR2, and CDR3). The domains of the light chain variable region are alternatively referred to as LCDR1, LCDR2, and LCDR3, and the domains of the heavy chain variable region are alternatively referred to as HCDR1, HCDR2, and HCDR3.

[0055] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to CD73 is substantially free of antibodies that do not bind to CD73). Moreover, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0056] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present in minor amounts. The monoclonal antibodies of the present invention can be produced by hybridoma methods, phage display techniques, single lymphocyte gene cloning techniques, or by recombinant DNA methods. For example, monoclonal antibodies can be produced by hybridomas comprising B cells obtained from a transgenic non-human animal, e.g., a transgenic mouse or rat, whose genome comprises a human heavy chain transgene and a light chain transgene.

[0057] As used herein, the term "antigen-binding fragment" refers to an antibody fragment, such as, for example, a diabody, Fab, Fab', F(ab'), Fv fragment, disulfide-stabilized Fv fragment (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabody (ds diabody), single-chain antibody molecule (scFv), single domain antibody (sdab), scFv dimer (bivalent diabody), multispecific antibody formed from a portion of an antibody containing one or more CDRs, camelized single domain antibody, nanobody, domain antibody, bivalent domain antibody, or any other antibody fragment that binds to an antigen but does not contain the complete antibody structure. An antigen-binding fragment is capable of binding to the same antigen as the parent antibody or parent antibody fragment binds. According to certain embodiments, the antigen-binding fragment comprises a light chain variable region, a light chain constant region, and an Fd segment of a heavy chain. According to other particular embodiments, the antigen-binding fragment comprises Fab and F(ab').

[0058] As used herein, the term "single chain antibody" refers to a conventional single chain antibody in the field that comprises a heavy chain variable region and a light chain variable region linked by a short peptide of about 15 to about 20 amino acids. As used herein, the term "single domain antibody" refers to a conventional single domain antibody in the field that comprises a heavy chain variable region and a heavy chain constant region or comprises only a heavy chain variable region.

[0059] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody having an amino acid sequence corresponding to an antibody produced by a human made using any technique known in the art. This definition of a human antibody includes intact or full-length antibodies, fragments thereof, and / or antibodies comprising at least one human heavy and / or light chain polypeptide.

[0060] As used herein, the term "humanized antibody" refers to a non-human antibody that has been modified to increase its sequence homology to that of a human antibody such that the antigen-binding properties of the antibody are retained but its antigenicity in the human body is reduced.

[0061] As used herein, the term "chimeric antibody" refers to an antibody in which the amino acid sequences of the immunoglobulin molecule are derived from two or more species. Often the variable regions of both the light and heavy chains correspond to the variable regions of antibodies derived from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and capacity, while the constant regions correspond to the sequences of antibodies derived from another species of mammal (usually human) to avoid eliciting an immune response in that species.

[0062] As used herein, the term "multispecific antibody" refers to an antibody comprising a plurality of immunoglobulin variable domain sequences, where a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., on the same protein (or on the same subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes do not overlap or substantially do not overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., on different proteins (or on different subunits of a multimeric protein). In one embodiment, the multispecific antibody comprises a third, fourth, or fifth immunoglobulin variable domain. In one embodiment, the multispecific antibody is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

[0063] As used herein, the term "bispecific antibody" refers to a multispecific antibody that binds only two epitopes or two antigens. A bispecific antibody is characterized by a first immunoglobulin variable domain sequence that has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., on the same protein (or on the same subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., on different proteins (or on different subunits of a multimeric protein). In one embodiment, a bispecific antibody comprises a heavy chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a first epitope and a heavy chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a second epitope. In one embodiment, a bispecific antibody comprises a half antibody, or fragment thereof, having binding specificity for a first epitope and a half antibody, or fragment thereof, having binding specificity for a second epitope. In one embodiment, a bispecific antibody comprises an scFv, or fragment thereof, having binding specificity for a first epitope, or an scFv, or fragment thereof, having binding specificity for a second epitope. In one embodiment, the first epitope is located on CD73 and the second epitope is located on PD-1, PD-L1, TIM-3, LAG-3, CD47, CD3, apelin, claudin 18.2, DLL3, folate receptor alpha (FRα), TIP-1, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, and / or other tumor-associated immune suppressors or surface antigens.

[0064] As used herein, the term "CD73" refers to ecto-5'-nucleotidase (ecto-5'-NT or EC 3.1.3.5), a cell surface phosphatase that catalyzes the dephosphorylation of extracellular AMP to generate adenosine. Physiologically, CD73 is induced by hypoxia and controls inflammation at wound sites (Bours et al., Pharmacol Ther 2006; 112:358-404). Pathologically, CD73 is often found overexpressed in regulatory T (Treg) cells and tumor cells (Paul et al., Trends Immunol 2012; 33:231-237). Increased CD73 activity leads to the accumulation of adenosine in the tumor microenvironment (TME). By binding to the adenosine receptors A2a and A2b, adenosine suppresses both innate and adaptive immunity by modulating many immune cells, such as macrophages (Csoka et al., FASEB J 2012; 26:376-386), dendritic cells (Panther et al., Blood 2003; 101:3985-3990), natural killer cells (Hausler et al., Cancer Immunol Immunother 2011; 60:1405-1418), and effector T cells (Hoskin et al., Int J Oncol 2008; 32:527-535). It has therefore been hypothesized that adenosine-induced immunosuppression may be alleviated by inhibiting the activity of CD73 in the TME. Indeed, in vivo animal studies (Jin et al., Cancer Res 2010; 70:2245-2255; Stagg et al., Cancer Res 2011; 71:2890-2900) have shown that inhibiting CD73 activity suppresses tumor formation and growth, suggesting that CD73 is a promising target for cancer therapy. The term "human CD73" refers to CD73 derived from humans. An exemplary amino acid sequence of human CD73 is set forth in GenBank Accession No. P21589.1 (SEQ ID NO:281).

[0065] As used herein, an antibody that "specifically binds to CD73" is one that is greater than or equal to 1×10 -7M or less, preferably 1×10 -8 M or less, preferably 5×10 -9 M or less, 1×10 -9 M or less, 5×10 -10 M or less, or 1 x 10 -10 It refers to an antibody that binds to CD73, preferably human CD73, with a KD of less than or equal to M. The term "KD" refers to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and expressed as a molar concentration (M). KD values ​​for antibodies can be determined using methods in the art in light of the present disclosure. For example, the KD of an antibody can be determined using surface plasmon resonance, e.g., using a biosensor system, e.g., a Biacore® system, or using biolayer interferometry, e.g., an Octet RED96 system.

[0066] The smaller the KD value of an antibody, the higher the affinity with which the antibody binds to a target antigen.

[0067] According to a particular aspect, the present invention provides a method for producing a method for treating a pulmonary circulation comprising the steps of: (1) SEQ ID NOs: 89, 90, 91, 149, 150, and 151, respectively; (2) SEQ ID NOs: 41, 42, 43, 101, 102, and 103, respectively; (3) SEQ ID NOs: 44, 45, 46, 104, 105, and 106, respectively; (4) SEQ ID NOs: 47, 48, 49, 107, 108, and 109, respectively; (5) SEQ ID NOs: 50, 51, 52, 110, 111, and 112, respectively; (6) SEQ ID NOs: 53, 54, 55, 113, 114, and 115, respectively; (7) SEQ ID NOs: 56, 57, 58, 116, 117, and 118, respectively; (8) SEQ ID NOs: 59, 60, 61, 119, 120, and 121, respectively; (9) SEQ ID NOs: 62, 63, 64, 122, 123, and 124, respectively; (10) SEQ ID NOs: 65, 66, 67, 125, 126, and 127, respectively; (11) SEQ ID NOs: 68, 69, 70, 128, 129, and 130, respectively; (12) SEQ ID NOs: 71, 72, 73, 131, 132, and 133, respectively; (13) SEQ ID NOs: 74, 75, 76, 134, 135, and 136, respectively; (14) SEQ ID NOs: 77, 78, 79, 137, 138, and 139, respectively; (15) SEQ ID NOs: 80, 81, 82, 140, 141, and 142, respectively; (16) SEQ ID NOs: 83, 84, 85, 143, 144, and 145, respectively; (17) SEQ ID NOs: 86, 87, 88, 146, 147, and 148, respectively; (18) SEQ ID NOs: 92, 93, 94, 152, 153, and 154, respectively; (19) SEQ ID NOs: 95, 96, 97, 155, 156, and 157, respectively; or (20) SEQ ID NOs: 98, 99, 100, 158, 159, and 160, respectively; The present invention relates to an isolated monoclonal antibody or an antigen-binding fragment thereof, which specifically binds to CD73, preferably human CD73, and which comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of

[0068] According to a particular aspect, the present invention provides a method for producing a method for treating a pulmonary circulation comprising the steps of: (1) SEQ ID NOs: 209, 210, 211, 269, 270, and 271, respectively; (2) SEQ ID NOs: 161, 162, 163, 221, 222, and 223, respectively; (3) SEQ ID NOs: 164, 165, 166, 224, 225, and 226, respectively; (4) SEQ ID NOs: 167, 168, 169, 227, 228, and 229, respectively; (5) SEQ ID NOs: 170, 171, 172, 230, 231, and 232, respectively; (6) SEQ ID NOs: 173, 174, 175, 233, 234, and 235, respectively; (7) SEQ ID NOs: 176, 177, 178, 236, 237, and 238, respectively; (8) SEQ ID NOs: 179, 180, 181, 239, 240, and 241, respectively; (9) SEQ ID NOs: 182, 183, 184, 242, 243, and 244, respectively; (10) SEQ ID NOs: 185, 186, 187, 245, 246, and 247, respectively; (11) SEQ ID NOs: 188, 189, 190, 248, 249, and 250, respectively; (12) SEQ ID NOs: 191, 192, 193, 251, 252, and 253, respectively; (13) SEQ ID NOs: 194, 195, 196, 254, 255, and 256, respectively; (14) SEQ ID NOs: 197, 198, 199, 257, 258, and 259, respectively; (15) SEQ ID NOs: 200, 201, 202, 260, 261, and 262, respectively; (16) SEQ ID NOs: 203, 204, 205, 263, 264, and 265, respectively; (17) SEQ ID NOs: 206, 207, 208, 266, 267, and 268, respectively; (18) SEQ ID NOs: 212, 213, 214, 272, 273, and 274, respectively; (19) SEQ ID NOs: 215, 216, 217, 275, 276, and 277, respectively; or (20) SEQ ID NOs: 218, 219, 220, 278, 279, and 280, respectively; The present invention relates to an isolated monoclonal antibody or an antigen-binding fragment thereof, which specifically binds to CD73, preferably human CD73, and which comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of

[0069] In another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to one of SEQ ID NOs: 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37, or 39; or a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to one of SEQ ID NOs: 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38, or 40. According to a preferred embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof of the invention comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37, or 39, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38, or 40, respectively.

[0070] According to another particular aspect, the present invention comprises (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:33, and a light chain variable region having the polypeptide sequence of SEQ ID NO:34; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 17, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 18; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 20; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 23, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 24; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 31, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 32; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 35, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 36; (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 37, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 38; or (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 39, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40; The present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising:

[0071] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 41, 42, 43, 101, 102 and 103, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2.

[0072] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 44, 45, 46, 104, 105 and 106, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 3, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 4. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 3, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 4.

[0073] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 47, 48, 49, 107, 108 and 109, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:5, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:6. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6.

[0074] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 50, 51, 52, 110, 111 and 112, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 8.

[0075] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 53, 54, 55, 113, 114 and 115, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 9, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 10. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 9, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 10.

[0076] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 56, 57, 58, 116, 117 and 118, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 11, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 12. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 11, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 12.

[0077] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 59, 60, 61, 119, 120 and 121, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 13, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 14. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14.

[0078] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 62, 63, 64, 122, 123 and 124, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 15, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 16. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0079] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 65, 66, 67, 125, 126 and 127, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 17, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 18. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 17, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 18.

[0080] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 68, 69, 70, 128, 129 and 130, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 19, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 20. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO:20.

[0081] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 71, 72, 73, 131, 132 and 133, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 21, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 22. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 21, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 22.

[0082] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 74, 75, 76, 134, 135 and 136, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 23, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 24. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 23, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 24.

[0083] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 77, 78, 79, 137, 138 and 139, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 25, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 26. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 25, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 26.

[0084] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 80, 81, 82, 140, 141 and 142, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 27, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 28. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 27, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 28.

[0085] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 83, 84, 85, 143, 144 and 145, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 30. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30.

[0086] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 86, 87, 88, 146, 147 and 148, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 31, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 32. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 31, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 32.

[0087] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 89, 90, 91, 149, 150 and 151, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 33, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 34. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 33, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 34.

[0088] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 92, 93, 94, 152, 153 and 154, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 35, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 36. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 35, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 36.

[0089] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 95, 96, 97, 155, 156 and 157, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 37, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 38. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 37, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 38.

[0090] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 98, 99, 100, 158, 159 and 160, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 39, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 40. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 39, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40.

[0091] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 161, 162, 163, 221, 222 and 223, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2.

[0092] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 164, 165, 166, 224, 225 and 226, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:3, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:4. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4.

[0093] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 167, 168, 169, 227, 228 and 229, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:5, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:6. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6.

[0094] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 170, 171, 172, 230, 231 and 232, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 8. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 8.

[0095] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 173, 174, 175, 233, 234 and 235, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 9, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 10. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 9, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 10.

[0096] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 176, 177, 178, 236, 237 and 238, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 11, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 12. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 11, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 12.

[0097] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 179, 180, 181, 239, 240 and 241, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 13, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 14. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14.

[0098] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 182, 183, 184, 242, 243 and 244, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0099] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 185, 186, 187, 245, 246 and 247, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 17, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 18. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 17, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 18.

[0100] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 188, 189, 190, 248, 249 and 250, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 19, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 20. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO:20.

[0101] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 191, 192, 193, 251, 252 and 253, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 21, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 22. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 21, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 22.

[0102] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 194, 195, 196, 254, 255 and 256, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 23, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 24. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 23, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 24.

[0103] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 197, 198, 199, 257, 258 and 259, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 25, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 26. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 25, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 26.

[0104] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 200, 201, 202, 260, 261 and 262, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 27, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 28. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 27, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 28.

[0105] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 203, 204, 205, 263, 264 and 265, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 30. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30.

[0106] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 206, 207, 208, 266, 267 and 268, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 31, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 32. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 31, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 32.

[0107] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 209, 210, 211, 269, 270 and 271, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 33, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 34. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 33, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 34.

[0108] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 212, 213, 214, 272, 273 and 274, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 35, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 36. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 35, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 36.

[0109] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 215, 216, 217, 275, 276 and 277, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 37, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 38. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 37, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 38.

[0110] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 having the polypeptide sequences of SEQ ID NOs: 218, 219, 220, 278, 279 and 280, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 39, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 40. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 39, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40.

[0111] In another particular embodiment, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof of the present invention that inhibits the enzymatic activity of soluble and / or cell-surface CD73. CD73 is a cell-surface glycophosphatidylinositol (GPI)-anchored protein that can be released to yield a catalytically active form in solution (Arias et al., J Cell Biol. 1997; 136(2):421-31). Here, cell-surface CD73 refers to cell-surface CD73 anchored by GPI, and soluble CD73 refers to CD73 that is released from the cell surface and becomes catalytically active in solution.

[0112] In another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, which interferes (prevents or prevents) dimerization of CD73. As used herein, "dimerization" refers to the formation of a CD73 dimer by two subunits. Preventing dimerization of CD73 refers to blocking or inhibiting the formation of a CD73 dimer.

[0113] In another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, which induces internalization of CD73. As used herein, "internalization of CD73" refers to the translocation of CD73 protein from the surface of a cell to the interior region of the cell.

[0114] According to another particular aspect, the present invention relates to an isolated monoclonal antibody or an antigen-binding fragment thereof of the present invention, which is capable of activating T cells by inhibiting the nucleotidase activity of CD73 and thus reducing adenosine levels.

[0115] In another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, which is chimeric.

[0116] In another particular aspect, the present invention relates to an isolated monoclonal antibody or an antigen-binding fragment thereof of the present invention, which is human or humanized.

[0117] According to another particular aspect, the present invention provides a method for producing a method of making a semiconductor device comprising: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 282, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 287, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 288, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 289, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 295, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 296, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 297, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; or (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 298, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299. The present invention relates to an isolated humanized monoclonal antibody or antigen-binding fragment thereof comprising:

[0118] In another general aspect, the present invention relates to an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof of the present invention. It will be appreciated by those skilled in the art that the coding sequence of a protein can be altered (e.g., substituted, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Thus, it will be understood by those skilled in the art that the nucleic acid sequence encoding the monoclonal antibody or antigen-binding fragment thereof of the present invention can be altered without changing the amino acid sequence of the protein.

[0119] In another general aspect, the present invention relates to a vector comprising an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof of the present invention. In view of the present disclosure, any vector known to those skilled in the art can be used, for example, a plasmid, a cosmid, a phage vector, or a viral vector. In some embodiments, the vector is a recombinant expression vector, for example, a plasmid. The vector can include any element for establishing the conventional functions of an expression vector, for example, a promoter, a ribosome binding element, a terminator, an enhancer, a selection marker, and an origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. Many expression vectors capable of delivering nucleic acids to cells are known in the art and can be used herein to produce the antibody or antigen-binding fragment thereof in cells. According to embodiments of the present invention, the recombinant expression vector can be generated using conventional cloning techniques or artificial gene synthesis.

[0120] In another general aspect, the present invention relates to a host cell comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof of the present invention. In view of the present disclosure, any host cell known to the skilled artisan can be used for recombinant expression of an antibody or antigen-binding fragment thereof of the present invention. In some embodiments, the host cell is an E. coli TG1 or BL21 cell (e.g., for expression of scFv or Fab antibodies), a CHO-DG44 or CHO-K1 cell, or a HEK293 cell (e.g., for expression of full-length IgG antibodies). According to certain embodiments, the recombinant expression vector is transformed into the host cell by conventional methods, e.g., chemical transfection, heat shock, or electroporation, and the recombinant expression vector is stably integrated into the genome of the host cell such that the recombinant nucleic acid is effectively expressed.

[0121] In another general aspect, the invention relates to a method of producing a monoclonal antibody or antigen-binding fragment thereof of the invention, comprising culturing a cell comprising nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof under conditions to produce the monoclonal antibody or antigen-binding fragment thereof of the invention, and recovering the antibody or antigen-binding fragment thereof from the cell or cell culture (e.g., from the supernatant). The expressed antibody or antigen-binding fragment thereof can be harvested from the cells and purified according to conventional techniques known in the art and described herein.

[0122] Pharmaceutical Compositions In another general aspect, the invention relates to a pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof of the invention and a pharma- ceutical carrier. The term "pharmaceutical composition" as used herein means a product comprising an antibody of the invention together with a pharma- ceutical carrier. The antibodies of the invention and compositions comprising them are also useful in the manufacture of medicaments for the therapeutic applications mentioned herein.

[0123] As used herein, the term "carrier" refers to any excipient, diluent, filler, salt, buffer, stabilizer, compatibilizer, oil, lipid, lipid including vesicle, microsphere, liposomal encapsulation, or other material known in the art for use in pharmaceutical formulations. It will be understood that the characteristics of the carrier, excipient, or diluent will depend on the route of administration for a particular application. As used herein, the term "pharmaceutical acceptable carrier" refers to a non-toxic material that does not interfere with the efficacy of the composition according to the invention or the biological activity of the composition according to the invention. According to certain embodiments, any pharmaceutical acceptable carrier suitable for use in antibody pharmaceutical compositions in light of the present disclosure can be used in the present invention.

[0124] Formulation of pharma- ceutically active ingredients with pharma- ceutically acceptable carriers is known in the art, for example, as in Remington: The Science and Practice of Pharmacy (e.g., 21st Edition (2005), any subsequent editions). Non-limiting examples of additional ingredients include buffers, diluents, solvents, tonicity adjusters, preservatives, stabilizers, and chelating agents. One or more pharma- ceutically acceptable carriers can be used in formulating the pharmaceutical compositions of the present invention.

[0125] In one embodiment of the present invention, the pharmaceutical composition is a liquid formulation. A preferred example of a liquid formulation is an aqueous formulation, i.e. a formulation that contains water. The liquid formulation can include solutions, suspensions, emulsions, microemulsions, gels, etc. An aqueous formulation typically contains at least 50% w / w water, or at least 60%, 70%, 75%, 80%, 85%, 90%, or at least 95% w / w water.

[0126] In one embodiment, the pharmaceutical composition can be formulated as an injectable that can be injected, for example, via an injection device (e.g., a syringe or an infusion pump). The injection can be delivered, for example, subcutaneously, intramuscularly, intraperitoneally, intravitreally, or intravenously.

[0127] In another embodiment, the pharmaceutical composition is a solid formulation, for example a freeze-dried or spray-dried composition, which may be used as is or the physician or patient may add solvents and / or diluents before use.Solid dosage forms may include tablets, for example compressed tablets and / or coated tablets, and capsules (for example hard or soft gelatin capsules).The pharmaceutical composition may also be in the form of, for example, sachets, dragees, powders, granules, lozenges, or powders for reconstitution.

[0128] The dosage form may be immediate release (in which case it may include a water soluble or dispersible carrier) or it may be delayed release, sustained release, or modified release (in which case it may include a water insoluble polymer that modulates the dissolution rate of the dosage form in the gastrointestinal tract or subcutaneously).

[0129] In other embodiments, the pharmaceutical compositions may be delivered intranasally, bucally, or sublingually.

[0130] The pH of the aqueous formulation may be between pH 3 and pH 10. In one embodiment of the invention, the pH of the formulation is from about 7.0 to about 9.5. In another embodiment of the invention, the pH of the formulation is from about 3.0 to about 7.0.

[0131] In another embodiment of the invention, the pharmaceutical composition comprises a buffering agent. Non-limiting examples of buffering agents include arginine, aspartic acid, bicine, citrate, disodium hydrogen phosphate, fumaric acid, glycine, glycylglycine, histidine, lysine, maleic acid, malic acid, sodium acetate, sodium carbonate, sodium dihydrogen phosphate, sodium phosphate, succinate, tartaric acid, tricine, and tris(hydroxymethyl)-aminomethane, and mixtures thereof. The buffering agents may be present individually or collectively in a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each of these specific buffering agents constitute alternative embodiments of the invention.

[0132] In another embodiment of the invention, the pharmaceutical composition comprises a preservative. Non-limiting examples of preservatives include benzethonium chloride, benzoic acid, benzyl alcohol, bronopol, butyl 4-hydroxybenzoate, chlorobutanol, chlorocresol, chlorhexidine, chlorphenesin, o-cresol, m-cresol, p-cresol, ethyl 4-hydroxybenzoate, imidurea, methyl 4-hydroxybenzoate, phenol, 2-phenoxyethanol, 2-phenylethanol, propyl 4-hydroxybenzoate, sodium dehydroacetate, thiomerosal, and mixtures thereof. The preservatives may be present individually or collectively in a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each of these specific preservatives constitute alternative embodiments of the invention.

[0133] In another embodiment of the present invention, the pharmaceutical composition comprises an isotonicity agent. Non-limiting examples of isotonicity agents include salts (e.g., sodium chloride), amino acids (e.g., glycine, histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, and threonine), alditols (e.g., glycerol, 1,2-propanediol propylene glycol), 1,3-propanediol, and 1,3-butanediol), polyethylene glycols (e.g., PEG400), and mixtures thereof. Another example of an isotonicity agent includes sugars. Non-limiting examples of sugars may be monosaccharides, disaccharides, or polysaccharides, or water-soluble glucans, such as fructose, glucose, mannose, sorbose, xylose, maltose, lactose, sucrose, trehalose, dextran, pullulan, dextrin, cyclodextrin, alpha and beta-HPCD, soluble starch, hydroxyethyl starch, and sodium carboxymethylcellulose. Another example of an isotonicity agent is a sugar alcohol, where the term "sugar alcohol" is defined as a C(4-8) hydrocarbon having at least one -OH group. Non-limiting examples of sugar alcohols include mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, and arabitol. The isotonicity agents, individually or collectively, may be present in a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions containing each of these specific isotonicity agents constitute alternative embodiments of the invention.

[0134] In another embodiment of the invention, the pharmaceutical composition comprises a chelating agent. Non-limiting examples of chelating agents include salts of citric acid, aspartic acid, ethylenediaminetetraacetic acid (EDTA), and mixtures thereof. The chelating agents may be present individually or collectively in a concentration of from about 0.01 mg / ml to about 50 mg / ml, e.g., from about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each of these specific chelating agents constitute alternative embodiments of the invention.

[0135] In another embodiment of the invention, the pharmaceutical composition comprises a stabilizer, non-limiting examples of which include one or more aggregation inhibitors, one or more oxidation inhibitors, one or more surfactants, and / or one or more protease inhibitors.

[0136] In another embodiment of the invention, the pharmaceutical composition comprises a stabilizer, said stabilizer being carboxy / hydroxycellulose and its derivatives (e.g., HPC, HPC-SL, HPC-L and HPMC), cyclodextrin, 2-methylthioethanol, polyethylene glycol (e.g., PEG3350), polyvinyl alcohol (PVA), polyvinylpyrrolidone, salts (e.g., sodium chloride), sulfur-containing substances, e.g., monothioglycerol), or thioglycolic acid. The stabilizers may be present individually or collectively in a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each of these specific stabilizers constitute alternative embodiments of the invention.

[0137] In a further embodiment of the invention, the pharmaceutical composition comprises one or more surfactants, preferably a surfactant, at least one surfactant, or two different surfactants. The term "surfactant" refers to any molecule or ion composed of a water-soluble (hydrophilic) portion and a fat-soluble (lipophilic) portion. The surfactant may be selected, for example, from the group consisting of anionic surfactants, cationic surfactants, nonionic surfactants, and / or zwitterionic surfactants. The surfactants may be present, individually or collectively, in a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each of these specific surfactants constitute alternative embodiments of the invention.

[0138] In further embodiments of the invention, the pharmaceutical composition comprises one or more protease inhibitors, such as, for example, EDTA, and / or benzamidine hydrochloride (HCl). The protease inhibitors, individually or collectively, may be present in a concentration of from about 0.1 mg / ml to about 20 mg / mml. Pharmaceutical compositions comprising each of these specific protease inhibitors constitute alternative embodiments of the invention.

[0139] In another general aspect, the invention relates to a method for producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof of the invention, the method comprising the step of combining the monoclonal antibody or antigen-binding fragment thereof with a pharma- ceutically acceptable carrier to obtain the pharmaceutical composition.

[0140] How to use In another general aspect, the present invention relates to a method of inhibiting the nucleotidase activity of CD73 in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody that specifically binds CD73, or an antigen-binding fragment thereof, or a pharmaceutical composition of the invention.

[0141] In another general aspect, the present invention relates to a method of disrupting (preventing or preventing) dimerization of CD73 in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds CD73, or a pharmaceutical composition of the invention.

[0142] In another general aspect, the present invention relates to a method for inducing internalization of CD73 in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to CD73, or a pharmaceutical composition of the invention. Internalization of CD73 into cells results in a reduction in the cell surface content of CD73. The level of cell surface CD73 can be measured by methods known in the art, for example, immunohistochemistry.

[0143] The functional activity of antibodies and antigen-binding fragments thereof that bind to CD73 can be characterized by methods known in the art and described herein. Cells expressing CD73 are pre-incubated with the antibody, followed by measuring the cell surface CD73 nucleotidase activity in the presence of the antibody. Methods for characterizing antibodies and antigen-binding fragments thereof that bind to CD73 include affinity and specificity assays, including, but not limited to, Biacore, ELISA, and OctetRed analysis, and the functional activity of anti-CD73 mAbs can also be evaluated in nucleotidase activity assays. According to certain embodiments, methods for characterizing antibodies and antigen-binding fragments thereof that bind to CD73 include those described below.

[0144] In another general aspect, the present invention relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds CD73 or a pharmaceutical composition of the invention. The cancer may be selected from, for example, but not limited to, lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML), and other liquid tumors.

[0145] In another general aspect, the present invention relates to a method of treating an inflammatory disease in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds CD73, or a pharmaceutical composition of the invention.

[0146] According to an embodiment of the invention, the pharmaceutical composition comprises a therapeutically effective amount of an anti-CD73 antibody or antigen-binding fragment thereof. As used herein, the term "therapeutically effective amount" refers to an amount of an active ingredient or component that elicits a desired biological or medical response in a subject. The therapeutically effective amount can be determined empirically and routinely in relation to the stated purpose.

[0147] As used herein with reference to an anti-CD73 antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to an amount of an anti-CD73 antibody or antigen-binding fragment thereof that modulates a tumor reduction response in a subject in need thereof, and as used herein with reference to an anti-CD73 antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to an amount of an anti-CD73 antibody or antigen-binding fragment thereof that provides treatment of a disease, disorder, or condition, prevents or slows the progression of the disease, disorder, or condition, or reduces or completely alleviates symptoms associated with the disease, disorder, or condition.

[0148] According to certain embodiments, the disease, disorder or condition to be treated is a cancer, preferably selected from the group consisting of lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML), and other liquid tumors. According to other particular embodiments, the disease, disorder or condition to be treated is an inflammatory disease.

[0149] According to certain embodiments, a therapeutically effective amount refers to an amount of therapy sufficient to achieve one, two, three, four or more of the following effects: (i) reducing or ameliorating the severity of the disease, disorder or condition being treated, or the symptoms associated therewith; (ii) shortening the duration of the disease, disorder or condition being treated, or the symptoms associated therewith; (iii) preventing the progression of the disease, disorder or condition being treated, or the symptoms associated therewith; (iv) causing regression of the disease, disorder or condition being treated, or the symptoms associated therewith; (v) preventing the development or onset of the disease, disorder or condition being treated, or the symptoms associated therewith; (vi) preventing the recurrence of the disease, disorder or condition, or symptoms associated therewith, being treated; (vii) reducing hospitalization of a subject having the disease, disorder or condition, or symptoms associated therewith, being treated; (viii) shortening the length of hospitalization of a subject having the disease, disorder or condition, or symptoms associated therewith, being treated; (ix) increasing the survival rate of a subject having the disease, disorder or condition, or symptoms associated therewith, being treated; (xi) inhibiting or reducing the disease, disorder or condition, or symptoms associated therewith, being treated in a subject; and / or (xii) enhancing or improving the prophylactic or therapeutic efficacy of another therapy.

[0150] The therapeutically effective amount or therapeutically effective dosage may vary depending on a variety of factors, such as the disease, disorder or condition being treated, the means of administration, the target site, the physiological state of the subject (including, for example, age, weight, health), whether the subject is human or animal, other medications being administered, and whether the treatment is prophylactic or therapeutic. Treatment dosages are optimally titrated to optimize safety and efficacy.

[0151] According to certain embodiments, the compositions described herein are formulated to be suitable for the intended route of administration to a subject. For example, the compositions described herein can be formulated to be suitable for intravenous, subcutaneous, or intramuscular administration.

[0152] As used herein, the terms "treat", "treating" and "treatment" are all intended to refer to an improvement or reversal of at least one measurable physical parameter associated with a cancer and / or inflammatory disease, disorder or condition, which may, but is not necessarily discernible in the subject. The terms "treat", "treating" and "treatment" may also refer to causing regression of, preventing the progression of, or at least slowing the rate of progression of, a disease, disorder or condition. In certain embodiments, "treat", "treating" and "treatment" refer to alleviating, preventing the development or onset of, or shortening the duration of, one or more symptoms associated with a disease, disorder or condition, e.g., a tumor or, more preferably, a cancer. In certain embodiments, "treat", "treating" and "treatment" refer to preventing the recurrence of a disease, disorder or condition. In certain embodiments, "treat," "treating," and "treatment" refer to increasing the survival rate of a subject having a disease, disorder, or condition. In certain embodiments, "treat," "treating," and "treatment" refer to the elimination of a disease, disorder, or condition in a subject.

[0153] According to certain embodiments, the compositions of the present invention are used in the treatment of cancer and / or inflammatory diseases, disorders or conditions. It may be used in combination with another treatment in cancer therapy, including but not limited to chemotherapy, anti-CD20 mAb, anti-TIM-3 mAb, anti-LAG-3 mAb, anti-CD47 mAb, anti-apelin mAb, anti-claudin 18.2 mAb, anti-DLL3 mAb, anti-FRα (folate receptor alpha) mAb, anti-TIP-1 mAb, anti-CTLA-4 antibody, anti-PD-L1 antibody, anti-PD-1 antibody, PD-1 / PD-L1 therapy, other cancer immunotherapy, anti-angiogenesis drug, radiation therapy, antibody-drug conjugate (ADC), targeted therapy, or other anti-cancer drug. Anti-CD73 antibodies can be used to construct bispecific antibodies with partner mAbs against PD-1, PD-L1, LAG3, TIM-3, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, CD47, CD3, apelin, claudin 18.2, DLL3, TIP-1, folate receptor alpha (FRα), and / or other tumor surface antigens to treat cancers / tumors expressing both CD73 and a specific tumor associated antigen.

[0154] As used herein, the term "in combination" in the context of administering two or more treatments to a subject refers to the use of two or more treatments. The use of the term "in combination" does not limit the order in which treatments are administered to a subject. For example, a first treatment (e.g., a composition described herein) can be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) administration of a second treatment to a subject.

[0155] In another general aspect, the invention relates to a method of determining the level of CD73 in a subject, the method comprising the steps of: (a) obtaining a sample from the subject; (b) contacting the sample with a monoclonal antibody or antigen-binding fragment thereof of the invention; and (c) determining the level of CD73 in the subject.

[0156] In another general aspect, the present invention relates to a method for determining the ecto-5'-nucleotidase activity of CD73 in a subject, wherein the enzymatic activity can be completely inhibited by a monoclonal antibody or antigen-binding fragment thereof of the invention, comprising the steps of: (a) obtaining a sample from a subject; (b) contacting the sample with an antibody or antigen-binding fragment thereof of the invention; and (c) determining the ecto-5'-nucleotidase activity of CD73 in the subject.

[0157] As used herein, "sample" refers to a biological sample isolated from a subject, including, but not limited to, whole blood, serum, plasma, blood cells, endothelial cells, tissue biopsy (e.g., cancer tissue), lymphatic fluid, ascites fluid, interstitial fluid, bone marrow, cerebrospinal fluid, saliva, mucus, sputum, sweat, urine, or any other secretion, excretion, or other bodily fluid. A "blood sample" refers to whole blood or any fraction thereof, including blood cells, serum, and plasma.

[0158] In certain embodiments, the level of CD73 in a subject can be determined using an assay selected from, but not limited to, Western blot assay, ELISA assay, and / or immunohistochemistry (IHC). Relative protein levels can be determined by using Western blot analysis and IHC, and absolute protein levels can be determined using ELISA assay. When determining the relative level of CD73, the level of CD73 can be determined between at least two samples, for example, between samples at different time points from the same subject, between samples from different tissues of the same subject, and / or between samples from different subjects. Alternatively, when the absolute level of CD73 is determined, for example, by ELISA assay, the absolute level of CD73 in a sample can be determined by making a standard for the ELISA assay before testing the sample. A person skilled in the art will understand which analytical technique to use to determine the level of CD73 in a sample from a subject using the antibody or antigen-binding fragment thereof of the present invention.

[0159] In certain embodiments, the ecto-5'-nucleotidase activity of CD73 in a subject can be determined using assays known in the art, see, for example, Geoghegan et al., MAbs 8:454-67 (2016) and Hay et al., Oncoimmunology Aug. 5(8):e1208875 (2016). A person skilled in the art will know which analytical techniques to use to determine the ecto-5'-nucleotidase activity of CD73 in a sample from a subject.

[0160] By utilizing a method for determining the level of CD73 or the level of CD73 ecto-5'-nucleotidase activity in a sample from a subject, a diagnosis of abnormal (elevated, reduced or insufficient) CD73 level and / or activity in a disease can be led to make appropriate therapeutic decisions. Such diseases can be selected from, but are not limited to, cancer and / or inflammatory diseases. Furthermore, by monitoring the level and / or activity of CD73 in a subject, the risk of developing the above-mentioned diseases can be determined based on knowledge of the level and / or activity of CD73 in a particular disease and / or during the progression of a particular disease.

[0161] Embodiment The present invention also provides the following non-limiting embodiments.

[0162] In embodiment 1, a. SEQ ID NOs: 89, 90, 91, 149, 150, and 151, respectively; b. SEQ ID NOs: 41, 42, 43, 101, 102, and 103, respectively; c. SEQ ID NOs: 44, 45, 46, 104, 105, and 106, respectively; d. SEQ ID NOs: 47, 48, 49, 107, 108, and 109, respectively; e. SEQ ID NOs: 50, 51, 52, 110, 111, and 112, respectively; f. SEQ ID NOs:53, 54, 55, 113, 114, and 115, respectively; g. SEQ ID NOs: 56, 57, 58, 116, 117, and 118, respectively; h. SEQ ID NOs: 59, 60, 61, 119, 120, and 121, respectively; i. SEQ ID NOs: 62, 63, 64, 122, 123, and 124, respectively; j. SEQ ID NOs: 65, 66, 67, 125, 126, and 127, respectively; k. SEQ ID NOs: 68, 69, 70, 128, 129, and 130, respectively; l. SEQ ID NOs: 71, 72, 73, 131, 132 and 133, respectively; m. SEQ ID NOs: 74, 75, 76, 134, 135, and 136, respectively; n. SEQ ID NOs: 77, 78, 79, 137, 138 and 139, respectively; o. SEQ ID NOs: 80, 81, 82, 140, 141 and 142, respectively; p. SEQ ID NOs: 83, 84, 85, 143, 144 and 145, respectively; q. SEQ ID NOs: 86, 87, 88, 146, 147 and 148, respectively; r. SEQ ID NOs: 92, 93, 94, 152, 153, and 154, respectively; s. SEQ ID NOs: 95, 96, 97, 155, 156, and 157, respectively; or t. SEQ ID NOs: 98, 99, 100, 158, 159 and 160, respectively; and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0163] In embodiment 2, a. SEQ ID NOs: 209, 210, 211, 269, 270, and 271, respectively; b. SEQ ID NOs: 161, 162, 163, 221, 222, and 223, respectively; c. SEQ ID NOs: 164, 165, 166, 224, 225, and 226, respectively; d. SEQ ID NOs: 167, 168, 169, 227, 228, and 229, respectively; e. SEQ ID NOs: 170, 171, 172, 230, 231, and 232, respectively; f. SEQ ID NOs: 173, 174, 175, 233, 234, and 235, respectively; g. SEQ ID NOs: 176, 177, 178, 236, 237, and 238, respectively; h. SEQ ID NOs: 179, 180, 181, 239, 240, and 241, respectively; i. SEQ ID NOs: 182, 183, 184, 242, 243, and 244, respectively; j. SEQ ID NOs: 185, 186, 187, 245, 246, and 247, respectively; k. SEQ ID NOs: 188, 189, 190, 248, 249, and 250, respectively; l. SEQ ID NOs: 191, 192, 193, 251, 252 and 253, respectively; m. SEQ ID NOs: 194, 195, 196, 254, 255, and 256, respectively; n. SEQ ID NOs: 197, 198, 199, 257, 258 and 259, respectively; o. SEQ ID NOs: 200, 201, 202, 260, 261 and 262, respectively; p. SEQ ID NOs: 203, 204, 205, 263, 264 and 265, respectively; q. SEQ ID NOs: 206, 207, 208, 266, 267, and 268, respectively; r. SEQ ID NOs: 212, 213, 214, 272, 273, and 274, respectively; s. SEQ ID NOs: 215, 216, 217, 275, 276, and 277, respectively; or t. SEQ ID NOs: 218, 219, 220, 278, 279, and 280, respectively; and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0164] Embodiment 3 is an isolated monoclonal antibody or antigen-binding fragment of embodiment 1 or 2, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37, or 39, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38, or 40.

[0165] In embodiment 4, (a) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 33, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 34; (b) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2; (c) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4; (d) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6; (e) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8; (f) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10; (g) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12; (h) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; (i) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (j) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 17, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 18; (k) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 20; (l) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22; (m) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:23, and a light chain variable region having the polypeptide sequence of SEQ ID NO:24; (n) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26; (o) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28; (p) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30; (q) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 31, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 32; (r) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 35, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 36; (s) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 37, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 38; or (t) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 39, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40. The isolated monoclonal antibody or antigen-binding fragment of any one of embodiments 1 to 3, comprising:

[0166] Embodiment 5 is an isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 4, which inhibits the enzymatic activity of soluble and / or cell surface CD73.

[0167] Embodiment 6 is an isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 4, which interferes with dimerization of CD73.

[0168] Embodiment 7 is an isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 4, which induces internalization of CD73.

[0169] Embodiment 8 is the isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 7, which is chimeric.

[0170] Embodiment 9 is an isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 8, which is human or humanized.

[0171] Embodiment 10 is (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:286, and a light chain variable region having the polypeptide sequence of SEQ ID NO:293; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 282, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 287, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 288, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 289, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 295, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 296, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 297, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; or (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 298, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1 to 9, comprising:

[0172] Embodiment 11 is an isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 10, which is capable of activating T cells.

[0173] Embodiment 12 is an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 11.

[0174] Embodiment 13 is a vector comprising the isolated nucleic acid of embodiment 12.

[0175] Embodiment 14 is a host cell comprising the vector of embodiment 13.

[0176] Embodiment 15 is a pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 11 and a pharma- ceutically acceptable carrier.

[0177] Embodiment 16 is a method of inhibiting the nucleotidase activity of CD73 in a subject in need thereof, comprising administering the pharmaceutical composition of embodiment 15 to the subject.

[0178] Embodiment 17 is a method of preventing dimerization of CD73 in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 15.

[0179] Embodiment 18 is a method of inducing internalization of CD73 in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 15.

[0180] Embodiment 19 is a method of treating cancer in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of embodiment 15.

[0181] Embodiment 20 is a method for producing the monoclonal antibody or antigen-binding fragment thereof of any one of Embodiments 1 to 11, comprising the steps of culturing a cell containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions for producing the monoclonal antibody or antigen-binding fragment, and recovering the antibody or antigen-binding fragment from the cell or culture.

[0182] Embodiment 21 is a method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 11, comprising combining the monoclonal antibody or antigen-binding fragment with a pharma- ceutical acceptable carrier to obtain the pharmaceutical composition.

[0183]

[0023] Embodiment 22 is a method for determining a level of CD73 in a subject, comprising: a. obtaining a sample from a subject; b. contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-11; and c. Determining the level of CD73 in the subject The method includes:

[0184] Embodiment 23 is the method of embodiment 22, wherein the sample is a tissue sample.

[0185] Embodiment 24 is the method of embodiment 23, wherein the tissue sample is a cancer tissue sample.

[0186] Embodiment 25 is the method of embodiment 22, wherein the sample is a blood sample.

[0187] Embodiment 26 is a method for determining the ecto-5'-nucleotidase activity of CD73 in a subject, comprising: a. obtaining a sample from a subject; b. contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1-11; and c. Determining the ecto-5'-nucleotidase activity of CD73 in the subject. The method includes:

[0188] Embodiment 27 is the method of embodiment 26, wherein the sample is a tissue sample.

[0189] Embodiment 28 is the method of embodiment 27, wherein the tissue sample is a cancer tissue sample.

[0190] Embodiment 29 is the method of embodiment 26, wherein the sample is a blood sample. EXAMPLES

[0191] [Example 1] Identification of anti-CD73 monoclonal antibodies Mice were immunized with a mixture of human CD73 (huCD73-HIS; containing residues 27-547) and mouse CD73 (mCD73-HIS; containing residues 29-549) fusion proteins, both containing a C-terminal HIS tag. Plasma titers were determined by ELISA. After euthanasia, spleens and lymph nodes were harvested and hybridomas were generated. Hybridomas were grown in 384-well tissue culture plates and supernatants from individual wells were screened by ELISA using huCD73-HIS and by fluorescence-activated cell sorting (FACS) using MDA-MB-231 cells. Positive clones were further analyzed by nucleotidase activity assay with CD73 immobilized on the plate. Top positive clones that showed inhibition of nucleotidase activity were isolated and sequenced.

[0192] The sequences of the heavy and light chain variable regions for anti-CD73 monoclonal antibodies are provided in Tables 1 and 2, and the CDR regions for anti-CD73 monoclonal antibodies are provided in Tables 3-6.

[0193] [Table 1] JPEG0007636483000002.jpg230159JPEG0007636483000003.jpg20159

[0194] [Table 2] JPEG0007636483000005.jpg23159

[0195] [Table 3]

[0196] [Table 4] JPEG0007636483000008.jpg65156

[0197] [Table 5]

[0198] [Table 6]

[0199] [Example 2] Production and purification of mAb from culture medium of transfected 293E cells To obtain recombinant anti-CD73 chimeric mAb, expression vectors containing mouse variable regions (VH and VL) fused to human IgG1 heavy and kappa light chain constant regions, respectively, with LALA mutations (L234A / L235A) were transiently transfected into 293E cells. Recombinant antibodies produced in suspension of 293E cells were purified using Protein A affinity chromatography.

[0200] [Example 3] Inhibition of the nucleotidase activity of immobilized CD73 by purified anti-CD73 chimeric mAb HIS-tagged human CD73 (huCD73-HIS, 60 μL / well at 0.15 μg / mL) in assay buffer (25 mM Tris-HCl, pH 7.5, and 5 mM MgCl2) was coated onto nickel-coated plates (ThermoFisher Scientific, Cat. No. 15242) overnight at 4°C. After washing, anti-CD73 chimeric mAb was added and incubated at room temperature for 1 h. The enzymatic reaction measuring the nucleotidase activity of CD73 was initiated by adding 100 μM AMP to the plate. The reaction was allowed to proceed for 20 min at 37°C. Free phosphate was detected and quantified by Malachite Green Phosphate Detection Kit (R&D Systems, Cat. No. DY996). Inhibition of the nucleotidase activity of immobilized CD73 by anti-CD73 chimeric mAb is shown in Figures 1A and 1B. No antibody is the enzyme reaction where no inhibitor was added; AMPCP (adenosine 5'(α,β-methylene)diphosphate) was used as a control for inhibition of enzyme activity.

[0201] [Example 4] ELISA binding assay using chimeric mAb HIS-tagged human CD73 in carbonate coating buffer (huCD73-HIS, 50 μL / well (85 nM) at 5 μg / mL) was coated onto ELISA plates overnight at 4° C. After washing with TBST buffer (TBS buffer with 0.05% Tween 20), the ELISA plates were blocked with 5% BSA in TBST for 1 h at room temperature and washed again. Chimeric anti-CD73 antibody was added, mixed, and incubated for 1 h at room temperature. The plates were washed and binding of the anti-CD73 antibody to the immobilized huCD73-HIS was detected by adding anti-human IgG conjugated to horseradish peroxidase (anti-hIgG-HRP) (ThermoFisher Scientific, Cat. No. H10007) and incubating for 1 h. Plates were washed and the ELISA was developed using One-step Detection Solution (ThermoFisher Scientific, Cat. No. 34029) and measured as absorbance at 450 nm. The results for binding of anti-CD73 mAb to CD73 are shown in Figures 2A-2E.

[0202] [Example 5] Inhibition of nucleotidase activity of soluble CD73 by anti-CD73 mAb The assay was performed using recombinant human CD73 in solution, not immobilized on plates. HIS-tagged human CD73 (huCD73-HIS) (BPS Bioscience, Cat. No. 71184) at a final concentration of 0.5 nM and various concentrations of anti-CD73 antibodies were incubated in assay buffer (25 mM Tris-HCl, pH 7.5, 5 mM MgCl2, 140 mM NaCl, and 0.1% BSA) for 20 min at 37°C. The enzymatic reaction was initiated by adding AMP (Sigma Aldrich, Cat. No. A2252) to a final concentration of 400 μM. After incubating the plate for 20 min at 37°C, the concentration of inorganic phosphate was determined by Malachite Green Phosphate Detection Kit (R&D Systems, Cat. No. DY996). Two chimeric anti-CD73 mAbs, 60A9A / IgG4 and 39G8A / IgG4, were generated by fusing the VH and VL regions of murine mAbs 60A9A and 39G8A to the constant regions of human IgG4 heavy chain and kappa light chain, respectively. The chimeric antibodies were expressed in CHO cells, purified using Protein A affinity chromatography, and tested in this assay. The results for inhibition of the nucleotidase activity of soluble CD73 by 60A9A / IgG4 and 39G8A / IgG4 are shown in FIG. 3.

[0203] [Example 6] Humanization of anti-CD73 mAb The murine anti-CD73 mAbs 60A9A and 39G8A were humanized to reduce potential immunogenicity when used in human patients. The sequences of the variable regions of the heavy and light chains (VH and VL) were compared to human antibody sequences in the Protein Data Bank (PDB) database and a homology model was constructed. The CDRs of both the heavy and light chains of the murine mAbs were grafted into human frameworks most likely to maintain the correct structure required for antigen binding. The sequences of the humanized VH and VL regions are shown in Tables 7 and 8.

[0204] The humanized VH and VL regions were fused to human IgG1 heavy and kappa light chain constant regions with LALA mutations (L234A / L235A), respectively. Constructs corresponding to the mAb sequences were used for transient transfection in 293E cells, and the humanized mAbs were purified using Protein A chromatography. The humanized mAbs were tested in an enzyme activity assay using soluble human CD73. In this assay, 0.4 nM huCD73 was incubated with the mAbs for 1 h on ice, after which the enzyme reaction was started by adding AMP to a final concentration of 100 μM. The IC50 values ​​for the humanized mAbs are shown in Table 9.

[0205] Humanized mAb 60A9-H5L4 in a human IgG4 / kappa backbone (60A9-H5L4 / IgG4) was analyzed in an ELISA binding assay, with an IgG4 chimeric version of 60A9A (60A9A / IgG4) used as a control. The ELISA binding results are shown in Figure 4A. 60A9-H5L4 / IgG4 was also analyzed for its ability to inhibit the nucleotidase activity of soluble CD73 under the conditions of Example 5. The results are shown in Figure 4B.

[0206] [Table 7]

[0207] [Table 8] JPEG0007636483000013.jpg29160

[0208] [Table 9]

[0209] [Example 7] Inhibition of the nucleotidase activity of cell surface CD73 by anti-CD73 mAb A375 cells were harvested with PBS-EDTA (2 mM EDTA in PBS) and washed twice in PBS buffer supplemented with 0.1% BSA. 100,000 cells were plated in the presence of various concentrations of anti-CD73 antibodies and incubated at 37°C for 20 min. AMP (Sigma Aldrich, Catalog No. A2252) was then added to a final concentration of 125 μM in a final volume of 60 μL and the reaction was kept at 37°C for 60 min. The plate was then centrifuged and 40 μL of the supernatant was transferred to a new plate. ATP (Sigma Aldrich, Catalog No. A9187) was added to a final concentration of 100 μM in a final volume of 50 μL and the mixture was incubated at 37°C for 15 min. CellTiter-Glo® 2.0 reagent (Promega, catalog number G9242) was added at a 1:1 ratio to determine AMP levels in the mixtures, and the enzyme activity in a given reaction was calculated based on the AMP concentration at the end of the reaction. Results for inhibition of CD73 activity by humanized anti-CD73 mAb (in a human IgG1 LALA / kappa backbone) in a cell-based assay are shown in Table 10.

[0210] Humanized anti-CD73 mAbs 60A9-H5L4 / IgG4, 60A9-H3L5 / IgG4, 60A9-H5L5 / IgG4, 60A9-H6L5 / IgG4, 60A9-H7L5 / IgG4, 60A9-H8L5 / IgG4 and 60A9-H9L5 / IgG4 were also tested in a cell-based assay using phosphate detection. In this assay, 50,000 A375 cells were washed and plated in assay buffer (25 mM Tris-HCl, pH 7.5, 5 mM MgCl2, 140 mM NaCl and 0.1% BSA) and the enzymatic reaction was continued for 20 min at 37° C. with a final AMP concentration of 400 μM. Free phosphate generated by hydrolysis of AMP was quantified using a Malachite Green Phosphate Detection Kit (R&D Systems, Cat. No. DY996). The results of this assay are shown in Figures 4C and 4D.

[0211] [Table 10]

[0212] [Example 8] Inhibition of the nucleotidase activity of CD73 in patient serum samples by anti-CD73 mAb Serum samples from patients with colorectal cancer were used to evaluate the inhibitory activity of humanized anti-CD73 mAb 60A9-H5L4 / IgG4. 20 μL of serum and 10 μL of anti-CD73 mAb were incubated for 30 min at 37° C. in PBS buffer supplemented with 0.1% BSA. 10 μL of 500 μM AMP (Sigma Aldrich, Catalog No. A2252) was added and the reaction mixture was incubated for 30 min at 37° C. ATP (Sigma Aldrich, Catalog No. A9187) was then added to a final concentration of 100 μM. 25 μL of the reaction mixture was transferred to a white half-area 96-well plate. CellTiter-Glo® 2.0 reagent (Promega, Catalog No. G9242) was added to the mixture in a 1:1 ratio and the residual AMP level was determined by measuring the inhibitory effect on ATP detection. A sample containing 100 μM AMP and 100 μM ATP was used to establish the assay background, and a sample containing only 100 μM ATP was used as a positive control.

[0213] Inhibition of the nucleotidase activity of CD73 in patient serum samples by humanized anti-CD73 mAb 60A9-H5L4 / IgG4 was analyzed at 1000 nM, and the results are shown in FIG.

[0214] [Example 9] Activation of T cell proliferation by humanized anti-CD73 mAb 60A9-H5L4 / IgG4 Human primary CD4+ T cells were isolated from frozen peripheral blood mononuclear cells using a CD4+ T cell isolation kit (Miltenyi Biotec, Cat. No. 130-096-533). Isolated CD4+ T cells at a density of 1,000,000 cells per mL of DPBS were labeled using a CellTrace™ CFSE cell proliferation kit (Invitrogen, Cat. No. C34554) for 30 minutes at 37°C. Cells were washed three times with chilled DPBS and resuspended in AIM V medium (Gibco, Cat. No. 12055083). 100,000 cells per well were preincubated with various concentrations of 60A9-H5L4 / IgG4 for 1 hour at 37°C. T cells were activated by adding a T cell activation reagent, a combination of Dynabeads Human T-Activator CD3 / CD28 (Gibco, Cat. No. 11131D) at a 1:1 bead-to-cell ratio, and human IL-2 to a final concentration of 60 IU / mL. AMP (Sigma Aldrich, Cat. No. A2252) was then added to a final concentration of 100 μM. After 72 h of incubation at 37°C, cells were stained with PE / Cy7 anti-human CD4 antibody (BioLegend, Cat. No. 357409). Both PE / Cy7 and CFSE signals were analyzed on an Attune NxT flow cytometer. As an assay window, CFSE CD4+ cells in the absence of T cell activation reagent did not undergo cell division and were therefore used to define the baseline, whereas activated CFSE CD4+ cells in the absence of AMP (treated with T cell activation reagent) underwent uninhibited cell division and were therefore used to define maximum proliferation. The effect of 60A9-H5L4 / IgG4 on activating T cell proliferation is shown in Figure 6. Cells treated with AMP were inhibited as expected, likely due to adenosine generation by cell surface CD73, and addition of 75 nM of anti-CD73 mAb 60A9-H5L4 / IgG4 activated the cells, suggesting that the mAb has functional activity in T cell activation.

[0215] An embodiment of the present invention will be described below. [1] a. SEQ ID NOs: 89, 90, 91, 149, 150, and 151, respectively; b. SEQ ID NOs: 41, 42, 43, 101, 102, and 103, respectively; c. SEQ ID NOs: 44, 45, 46, 104, 105, and 106, respectively; d. SEQ ID NOs: 47, 48, 49, 107, 108, and 109, respectively; e. SEQ ID NOs: 50, 51, 52, 110, 111, and 112, respectively; f. SEQ ID NOs:53, 54, 55, 113, 114, and 115, respectively; g. SEQ ID NOs: 56, 57, 58, 116, 117, and 118, respectively; h. SEQ ID NOs: 59, 60, 61, 119, 120, and 121, respectively; i. SEQ ID NOs: 62, 63, 64, 122, 123, and 124, respectively; j. SEQ ID NOs: 65, 66, 67, 125, 126, and 127, respectively; k. SEQ ID NOs: 68, 69, 70, 128, 129, and 130, respectively; l. SEQ ID NOs: 71, 72, 73, 131, 132 and 133, respectively; m. SEQ ID NOs: 74, 75, 76, 134, 135, and 136, respectively; n. SEQ ID NOs: 77, 78, 79, 137, 138 and 139, respectively; o. SEQ ID NOs: 80, 81, 82, 140, 141 and 142, respectively; p. SEQ ID NOs: 83, 84, 85, 143, 144 and 145, respectively; q. SEQ ID NOs: 86, 87, 88, 146, 147 and 148, respectively; r. SEQ ID NOs: 92, 93, 94, 152, 153, and 154, respectively; s. SEQ ID NOs: 95, 96, 97, 155, 156, and 157, respectively; or t. SEQ ID NOs: 98, 99, 100, 158, 159 and 160, respectively; and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of: [2] a. SEQ ID NOs: 209, 210, 211, 269, 270, and 271, respectively; b. SEQ ID NOs: 161, 162, 163, 221, 222, and 223, respectively; c. SEQ ID NOs: 164, 165, 166, 224, 225, and 226, respectively; d. SEQ ID NOs: 167, 168, 169, 227, 228, and 229, respectively; e. SEQ ID NOs: 170, 171, 172, 230, 231, and 232, respectively; f. SEQ ID NOs: 173, 174, 175, 233, 234, and 235, respectively; g. SEQ ID NOs: 176, 177, 178, 236, 237, and 238, respectively; h. SEQ ID NOs: 179, 180, 181, 239, 240, and 241, respectively; i. SEQ ID NOs: 182, 183, 184, 242, 243, and 244, respectively; j. SEQ ID NOs: 185, 186, 187, 245, 246, and 247, respectively; k. SEQ ID NOs: 188, 189, 190, 248, 249, and 250, respectively; l. SEQ ID NOs: 191, 192, 193, 251, 252 and 253, respectively; m. SEQ ID NOs: 194, 195, 196, 254, 255, and 256, respectively; n. SEQ ID NOs: 197, 198, 199, 257, 258 and 259, respectively; o. SEQ ID NOs: 200, 201, 202, 260, 261 and 262, respectively; p. SEQ ID NOs: 203, 204, 205, 263, 264 and 265, respectively; q. SEQ ID NOs: 206, 207, 208, 266, 267, and 268, respectively; r. SEQ ID NOs: 212, 213, 214, 272, 273, and 274, respectively; s. SEQ ID NOs: 215, 216, 217, 275, 276, and 277, respectively; or t. SEQ ID NOs: 218, 219, 220, 278, 279, and 280, respectively; and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of: [3] An isolated monoclonal antibody or an antigen-binding fragment thereof according to [1] or [2], comprising a heavy chain variable region having a polypeptide sequence that is at least 95% identical to SEQ ID NO: 33, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 35, 37 or 39, or a light chain variable region having a polypeptide sequence that is at least 95% identical to SEQ ID NO: 34, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 36, 38 or 40. [4] a. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 33, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 34; b. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:1, and a light chain variable region having the polypeptide sequence of SEQ ID NO:2; c. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:3, and a light chain variable region having the polypeptide sequence of SEQ ID NO:4; d. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6; e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:7, and a light chain variable region having the polypeptide sequence of SEQ ID NO:8; f. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:9, and a light chain variable region having the polypeptide sequence of SEQ ID NO:10; g. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:11, and a light chain variable region having the polypeptide sequence of SEQ ID NO:12; h. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:13, and a light chain variable region having the polypeptide sequence of SEQ ID NO:14; i. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:15, and a light chain variable region having the polypeptide sequence of SEQ ID NO:16; j. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:17, and a light chain variable region having the polypeptide sequence of SEQ ID NO:18; k. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 19, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 20; l. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:21, and a light chain variable region having the polypeptide sequence of SEQ ID NO:22; m. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:23, and a light chain variable region having the polypeptide sequence of SEQ ID NO:24; n. A heavy chain variable region having the polypeptide sequence of SEQ ID NO:25, and a light chain variable region having the polypeptide sequence of SEQ ID NO:26; o. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:27, and a light chain variable region having the polypeptide sequence of SEQ ID NO:28; p. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30; q. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:31, and a light chain variable region having the polypeptide sequence of SEQ ID NO:32; r. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:35, and a light chain variable region having the polypeptide sequence of SEQ ID NO:36; s. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:37, and a light chain variable region having the polypeptide sequence of SEQ ID NO:38; or t. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:39, and a light chain variable region having the polypeptide sequence of SEQ ID NO:40. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to [3], [5] An isolated monoclonal antibody or an antigen-binding fragment thereof according to any one of [1] to [4], which inhibits the enzymatic activity of soluble and / or cell surface CD73. [6] An isolated monoclonal antibody or an antigen-binding fragment thereof according to any one of [1] to [4], which inhibits dimerization of CD73. [7] An isolated monoclonal antibody or an antigen-binding fragment thereof according to any one of [1] to [4], which induces internalization of CD73. [8] An isolated monoclonal antibody or an antigen-binding fragment thereof according to any one of [1] to [7], which is a chimeric antibody. [9] An isolated monoclonal antibody or an antigen-binding fragment thereof according to any one of [1] to [8], which is human or humanized.

[10] (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:290; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO:282, and a light chain variable region having the polypeptide sequence of SEQ ID NO:291; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 282, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 283, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 285, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 293; (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 290; (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 291; (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 286, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 292; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 287, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 288, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 289, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 284, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 295, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 296, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 297, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299; or (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 298, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 299. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to [9],

[11] The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to

[10] , which is capable of activating T cells.

[12] An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof described in any one of [1] to

[11] .

[13] A vector comprising the isolated nucleic acid described in

[12] .

[14] A host cell comprising the vector described in

[13] .

[15] A pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof described in any one of [1] to

[11] and a pharma- ceutical acceptable carrier.

[16] A method for inhibiting the nucleotidase activity of CD73 in a subject in need thereof, comprising the step of administering to the subject the pharmaceutical composition described in

[15] .

[17] A method for preventing dimerization of CD73 in a subject in need thereof, comprising the step of administering to the subject the pharmaceutical composition described in

[15] .

[18] A method for inducing internalization of CD73 in a subject in need thereof, comprising the step of administering to the subject the pharmaceutical composition described in

[15] .

[19] A method for treating cancer in a subject in need thereof, comprising administering to the subject the pharmaceutical composition described in

[15] .

[20] A method for producing the monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to

[11] , comprising the steps of culturing cells containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions for producing the monoclonal antibody or antigen-binding fragment, and recovering the antibody or antigen-binding fragment from the cells or culture.

[21] A method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment according to any one of [1] to

[11] , comprising the steps of blending the monoclonal antibody or antigen-binding fragment with a pharma- ceutical carrier to obtain a pharmaceutical composition.

[22] A method for determining a level of CD73 in a subject, comprising: a. obtaining a sample from a subject; b. contacting a sample with the isolated monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to

[11] ; and c. Determining the level of CD73 in the subject The method includes:

[23] The method according to

[22] , wherein the sample is a tissue sample.

[24] The method according to

[23] , wherein the tissue sample is a cancer tissue sample.

[25] The method according to

[22] , wherein the sample is a blood sample.

[26] A method for determining ecto-5'-nucleotidase activity of CD73 in a subject, comprising the steps of: a. obtaining a sample from the subject; b. contacting a sample with the isolated monoclonal antibody or antigen-binding fragment thereof according to any one of [1] to

[11] ; and c. Determining the ecto-5'-nucleotidase activity of CD73 in the subject. The method includes:

[27] The method according to

[26] , wherein the sample is a tissue sample.

[28] The method according to

[27] , wherein the tissue sample is a cancer tissue sample.

[29] The method according to

[26] , wherein the sample is a blood sample. Those skilled in the art will recognize that changes may be made to the above-described embodiments without departing from the broad inventive concept thereof. It is understood, therefore, that the invention is not limited to the particular embodiments disclosed, but is intended to cover modifications within the spirit and scope of the invention as defined herein.

[0216] The specification includes the following sequence information: [Sequence table] SEQUENCE LISTING <110> Phanes Therapeutics, Inc. <120> Anti-CD73 Antibodies and Uses Thereof <130> PA23-380 <140> <141> 2019-03-05 <150> US 62 / 640,955 <151> 2018-03-09 <150> US 62 / 721,044 <151> 2018-08-22 <150> US 62 / 786,598 <151> 2018-12-31 <160> 299 <170> PatentIn version 3.5 <210> 1 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> 37C7A Heavy Chain Variable Region <400> 1 Gln Val Thr Leu Lys Glu Ser Gly Pro Gly Ile Leu Gln Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Phe Ser Gly Phe Ser Leu Ser Thr Phe 20 25 30 Gly Met Gly Val Thr Trp Ile Arg Gln Pro Ser Gly Lys Gly Leu Glu 35 40 45 Trp Leu Ala His Ile Trp Trp Asp Asp Asp Met Tyr Tyr Asn Pro Ala 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Phe Leu Lys Ile Ala Asn Val Asp Thr Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser Pro Ile Thr Thr Val Val Ala Asp Tyr Trp Gly Gln 100 105 110 Gly Ser Thr Leu Thr Val Ser Ser 115 120 <210> 2 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> 37C7A Light Chain Variable Region <400> 2 Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Leu Val His Ser 20 25 30 Tyr Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe 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 Leu Gly Val Tyr Phe Cys Ser Gln Asn 85 90 95 Thr His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Gln 100 105 110 <210> 3 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 38C16A Heavy Chain Variable Region <400> 3 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Tyr 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Phe Gly Ser Ser Tyr Arg Gly Pro Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 4 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> 38C16A Light Chain Variable Region <400> 4 Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 20 25 30 Asp Gly Arg Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 50 55 60 Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly 85 90 95 Thr His Phe Pro His Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 5 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 39G8A Heavy Chain Variable Region <400> 5 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Met Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asp Pro Ser Gly Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ser Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 6 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 39G8A Light Chain Variable Region <400> 6 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5 10 15 Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 7 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 43E8A Heavy Chain Variable Region <400> 7 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Asn Val Ile Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Arg Gly Asp Asp Gly Tyr Leu Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 8 <211> 111 <212> PRT <213> Artificial Sequence <220> <223> 43E8A Light Chain Variable Region <400> 8 Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Gln Gly Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His 65 70 75 80 Pro Met Glu Glu Asp Asp Thr Ala Met Tyr Phe Cys Gln Gln Ser Lys 85 90 95 Glu Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 9 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> 46J3A Heavy Chain Variable Region <400> 9 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Arg Ser Ser Ser Thr Val Tyr 65 70 75 80 Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Tyr Trp Asp Tyr Tyr Gly Ser Thr Tyr Gly Tyr Phe Asp Val 100 105 110 Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 10 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 46J3A Light Chain Variable Region <400> 10 Asp Val Gln Ile Thr Gln Ser Pro Ser Tyr Leu Ala Ala Ser Pro Gly 1 5 10 15 Glu Thr Ile Thr Ile Asn Cys Arg Ala Ser Lys Asn Ile Ser Lys Tyr 20 25 30 Leu Ala Trp Tyr Gln Glu Ile Pro Gly Lys Thr Tyr Asn Leu Leu Ile 35 40 45 Tyr Ser Gly Ser Thr Leu Gln Ser Gly Ile Pro Ser 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 Met Tyr Tyr Cys Gln Gln His Asn Glu Tyr Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 11 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> 46O19A Heavy Chain Variable Region <400> 11 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Asn Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asn Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Tyr Phe Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 12 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 46O19A Light Chain Variable Region <400> 12 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Glu Lys Pro Gly Gln 35 40 45 Pro Pro Lys Val Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 85 90 95 Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 13 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 47H24A Heavy Chain Variable Region <400> 13 Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Ser Tyr 20 25 30 Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Ser Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Ile 50 55 60 Ser Arg Leu Ser Ile Ser Lys Asp Asn Ser Lys Ser Gln Val Phe Phe 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asn Asp Thr Ala Ile Tyr Tyr Cys Ala 85 90 95 Ser Gln Tyr Val Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser 100 105 110 Thr <210> 14 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 47H24A Light Chain Variable Region <400> 14 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Arg 20 25 30 Leu Thr Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile 35 40 45 Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser 65 70 75 80 Glu Asp Phe Val Asp Tyr Phe Cys Leu Gln Tyr Ala Ser Ser Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 15 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 49O5A Heavy Chain Variable Region <400> 15 Gln Val Gln Leu Gln Gln Pro Gly Thr Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Lys Ser Asn Tyr Gly Tyr Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 16 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 49O5A Light Chain Variable Region <400> 16 Asp Ile Gln Met Thr Gln Ser Ser Ser Tyr Leu Ser Val Ser Leu Gly 1 5 10 15 Gly Arg Val Thr Ile Thr Cys Glu Ala Ser Asp His Ile Asp Asn Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Asn Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Thr Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Lys Asp Tyr Thr Leu Ser Ile Thr Ser Leu Gln Thr 65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Trp Ser Ser Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 17 <211> 124 <212> PRT <213> Artificial Sequence <220> <223> 50B10A Heavy Chain Variable Region <400> 17 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ser Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Gly Arg Val Tyr Tyr Asp Tyr Phe Tyr Ala Met Asp 100 105 110 Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 18 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 50B10A Light Chain Variable Region <400> 18 Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln 65 70 75 80 Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 19 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> 50H17A Heavy Chain Variable Region <400> 19 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Asp Tyr Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 20 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 50H17A Light Chain Variable Region <400> 20 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Val Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 85 90 95 Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 21 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> 50E16A Heavy Chain Variable Region <400> 21 Glu Leu Ala Arg Pro Trp Ala Ser Val Lys Ile Ser Cys Gln Ala Phe 1 5 10 15 Tyr Thr Phe Ser Arg Arg Met His Phe Ala Ile Arg Asp Thr Asn Tyr 20 25 30 Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Val Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Tyr Tyr Ser Asn Tyr Gly Gly Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 22 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 50E16A Light Chain Variable Region <400> 22 Asp Ile Val Met Thr Gln Phe His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Ser Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 23 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> 50F1A Heavy Chain Variable Region <400> 23 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ser Met His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Glu Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Asp Ile Phe Trp Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 24 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 50F1A Light Chain Variable Region <400> 24 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr 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 Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 85 90 95 Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 25 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> 54I14A Heavy Chain Variable Region <400> 25 Gln Ala Tyr Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Asn Met His Trp Val Lys Gln Thr Pro Arg Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Tyr Asp Tyr Asp Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 26 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 54I14A Light Chain Variable Region <400> 26 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Phe Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 27 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> 52K11A Heavy Chain Variable Region <400> 27 Glu Ile Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr 20 25 30 Asn Met Asn Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Asn Ile Asn Pro Tyr Tyr Gly Ser Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Asn Ser Leu Thr Ser Lys Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Gly Ser Ser Tyr Val Asp Tyr Ala Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Ser Val Thr Ser Val 115 <210> 28 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> 52K11A Light Chain Variable Region <400> 28 Asp Ile Val Met Thr Gln Ala Ala Ile Ser Asn Pro Val Thr Leu Gly 1 5 10 15 Thr Ser Ala Ser Ile Ser Cys Ser Ser Asn Lys Ser Leu Leu His Ser 20 25 30 Asn Asp Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Met 85 90 95 Leu Glu Arg Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 29 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> 53H21A Heavy Chain Variable Region <400> 29 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Ile Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe 50 55 60 Glu Gly Lys Ala Thr Leu Thr Ser Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Gly Asn Trp Asp Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 30 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> 53H21A Light Chain Variable Region <400> 30 Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 31 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> 53I12A Heavy Chain Variable Region <400> 31 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Pro Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Asn Ile Tyr Ala Gly Ser Ser Ser Ser Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly His Gly Tyr Asp Gly Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala 115 <210> 32 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 53I12A Light Chain Variable Region <400> 32 Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Lys Leu Pro Arg 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 33 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9A Heavy Chain Variable Region <400> 33 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Lys Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Ala Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 34 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9A Light Chain Variable Region <400> 34 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5 10 15 Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Asp Ser Gly Val 50 55 60 Pro Asp Arg Phe Ile Gly Gly Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 35 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> 39J9A Heavy Chain Variable Region <400> 35 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Leu Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Ile Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Gly Asp Tyr Tyr Gly Asn Pro Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 36 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 39J9A Light Chain Variable Region <400> 36 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ala Ala Ser Val Gly 1 5 10 15 Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Tyr Ser 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Asn Ala Asp Thr Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Met Lys Ile Asn Ser Met Gln Pro 65 70 75 80 Glu Asp Thr Ala Thr Tyr Phe Cys Lys Gln Ala Tyr Asp Val Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 37 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 49H8A Heavy Chain Variable Region <400> 37 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met Gln Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asp Pro Ser Asp Asn Tyr Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Glu Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Tyr Tyr Gly Tyr Ser Pro Ser Trp Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 38 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> 49H8A Light Chain Variable Region <400> 38 Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr 85 90 95 Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 39 <211> 117 <212> PRT <213> Artificial Sequence <220> <223> 51B10A Heavy Chain Variable Region <400> 39 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr 20 25 30 Tyr Met His Trp Val Lys Gln Ser His Val Lys Ser Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asn Pro Tyr Ser Gly Ala Thr Asn Ser Asn Gln Asn Phe 50 55 60 Lys Asp Lys Ala Ser Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu His Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Tyr Gly Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 40 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> 51B10A Light Chain Variable Region <400> 40 Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Ser Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35 40 45 Tyr Arg Ala Asn Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Glu Phe Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 41 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR1 <400> 41 Gly Phe Ser Leu Ser Thr Phe Gly Met Gly 1 5 10 <210> 42 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR2 <400> 42 Ile Trp Trp Asp Asp Asp Met 1 5 <210> 43 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR3 <400> 43 Ala Arg Ser Pro Ile Thr Thr Val Val Ala Asp Tyr 1 5 10 <210> 44 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR1 <400> 44 Gly Tyr Thr Phe Ser Ser Tyr Trp 1 5 <210> 45 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR2 <400> 45 Ile Leu Pro Gly Ser Gly Ser Thr 1 5 <210> 46 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR3 <400> 46 Ala Arg Gly Asp Tyr Phe Gly Ser Ser Tyr Arg Gly Pro Tyr 1 5 10 <210> 47 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR1 <400> 47 Gly Tyr Thr Phe Thr Ser Tyr Trp 1 5 <210> 48 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR2 <400> 48 Ile Asp Pro Ser Gly Gly Tyr Thr 1 5 <210> 49 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR3 <400> 49 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr 1 5 10 <210> 50 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR1 <400> 50 Gly Tyr Thr Phe Ser Asn Tyr Trp 1 5 <210> 51 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR2 <400> 51 Ile Leu Pro Gly Asn Val Ile Thr 1 5 <210> 52 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR3 <400> 52 Ala Arg Arg Gly Asp Asp Gly Tyr Leu Tyr Ala Met Asp Tyr 1 5 10 <210> 53 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR1 <400> 53 Gly Tyr Thr Phe Thr Asp Tyr Tyr 1 5 <210> 54 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR2 <400> 54 Ile Tyr Pro Gly Ser Gly Asn Thr 1 5 <210> 55 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR3 <400> 55 Ala Arg Tyr Trp Asp Tyr Tyr Gly Ser Thr Tyr Gly Tyr Phe Asp Val 1 5 10 15 <210> 56 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR1 <400> 56 Gly Tyr Thr Phe Thr Asp Tyr Asn 1 5 <210> 57 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR2 <400> 57 Ile Asn Pro Asn Asn Gly Gly Thr 1 5 <210> 58 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR3 <400> 58 Ala Arg Asp Tyr Phe Trp Tyr Phe Asp Val 1 5 10 <210> 59 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR1 <400> 59 Gly Phe Ser Leu Thr Ser Tyr Gly 1 5 <210> 60 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR2 <400> 60 Ile Trp Ser Gly Gly Ser Thr 1 5 <210> 61 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR3 <400> 61 Ala Ser Gln Tyr Val Ala Tyr 1 5 <210> 62 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR1 <400> 62 Gly Tyr Thr Phe Thr Ser Tyr Trp 1 5 <210> 63 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR2 <400> 63 Ile Asn Pro Ser Asn Gly Gly Thr 1 5 <210> 64 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR3 <400> 64 Ala Arg Ser Lys Ser Asn Tyr Gly Tyr Tyr Ala Met Asp Tyr 1 5 10 <210> 65 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR1 <400> 65 Gly Tyr Thr Phe Thr Ser Tyr Trp 1 5 <210> 66 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR2 <400> 66 Ile Asn Pro Ser Asn Gly Arg Thr 1 5 <210> 67 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR3 <400> 67 Ala Arg Ser Glu Gly Arg Val Tyr Tyr Asp Tyr Phe Tyr Ala Met Asp 1 5 10 15 Tyr <210> 68 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR1 <400> 68 Gly Tyr Ala Phe Thr Asn Tyr Leu 1 5 <210> 69 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR2 <400> 69 Ile Asn Pro Gly Ser Gly Gly Thr 1 5 <210> 70 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR3 <400> 70 Ala Arg Asp Tyr Tyr Trp Tyr Phe Asp Val 1 5 10 <210> 71 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR1 <400> 71 Ala Ile Arg Asp Thr Asn Tyr Trp 1 5 <210> 72 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR2 <400> 72 Ile Tyr Pro Gly Asn Gly Asp Thr 1 5 <210> 73 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR3 <400> 73 Ala Thr Tyr Tyr Ser Asn Tyr Gly Gly Ala Met Asp Tyr 1 5 10 <210> 74 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR1 <400> 74 Gly Tyr Thr Phe Thr Asp Tyr Ser 1 5 <210> 75 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR2 <400> 75 Ile Asn Thr Glu Thr Gly Glu Pro 1 5 <210> 76 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR3 <400> 76 Ala Arg Asp Ile Phe Trp Ala Met Asp Tyr 1 5 10 <210> 77 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR1 <400> 77 Gly Tyr Thr Phe Thr Ser Tyr Asn 1 5 <210> 78 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR2 <400> 78 Ile Tyr Pro Gly Asn Gly Asp Thr 1 5 <210> 79 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR3 <400> 79 Ala Arg Tyr Asp Tyr Asp Ala Met Asp Tyr 1 5 10 <210> 80 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR1 <400> 80 Gly Tyr Ser Phe Thr Gly Tyr Asn 1 5 <210> 81 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR2 <400> 81 Ile Asn Pro Tyr Tyr Gly Ser Thr 1 5 <210> 82 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR3 <400> 82 Ala Gly Ser Ser Tyr Val Asp Tyr Ala Met Asp Tyr 1 5 10 <210> 83 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR1 <400> 83 Gly Tyr Thr Phe Thr Ser Tyr Val 1 5 <210> 84 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR2 <400> 84 Ile Ile Pro Tyr Asn Asp Gly Thr 1 5 <210> 85 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR3 <400> 85 Ala Arg Trp Gly Asn Trp Asp Tyr Phe Asp Tyr 1 5 10 <210> 86 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR1 <400> 86 Gly Tyr Thr Phe Thr Ser Tyr Trp 1 5 <210> 87 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR2 <400> 87 Ile Tyr Ala Gly Ser Ser Ser Ser 1 5 <210> 88 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR3 <400> 88 Ala Arg Ser Gly His Gly Tyr Asp Gly Phe Ala Tyr 1 5 10 <210> 89 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR1 <400> 89 Gly Tyr Ala Phe Ser Ser Tyr Trp 1 5 <210> 90 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR2 <400> 90 Ile Tyr Pro Gly Asp Gly Asp Thr 1 5 <210> 91 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR3 <400> 91 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr 1 5 10 <210> 92 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR1 <400> 92 Gly Tyr Ala Phe Thr Asn Tyr Leu 1 5 <210> 93 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR2 <400> 93 Ile Asn Pro Gly Ser Gly Gly Thr 1 5 <210> 94 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR3 <400> 94 Ala Arg Arg Gly Asp Tyr Tyr Gly Asn Pro Phe Asp Tyr 1 5 10 <210> 95 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR1 <400> 95 Gly Tyr Thr Phe Thr Asn Tyr Trp 1 5 <210> 96 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR2 <400> 96 Ile Asp Pro Ser Asp Asn Tyr Thr 1 5 <210> 97 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR3 <400> 97 Ala Arg Gly Tyr Tyr Gly Tyr Ser Pro Ser Trp Phe Ala Tyr 1 5 10 <210> 98 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR1 <400> 98 Gly Tyr Ser Phe Thr Gly Tyr Tyr 1 5 <210> 99 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR2 <400> 99 Ile Asn Pro Tyr Ser Gly Ala Thr 1 5 <210> 100 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR3 <400> 100 Ala Arg Ser Tyr Tyr Gly Ala Met Asp Tyr 1 5 10 <210> 101 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR1 <400> 101 Gln Asn Leu Val His Ser Tyr Gly Asn Thr Tyr 1 5 10 <210> 102 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR2 <400> 102 Lys Val Ser 1 <210> 103 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR3 <400> 103 Ser Gln Asn Thr His Val Pro Trp Thr 1 5 <210> 104 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR1 <400> 104 Gln Ser Leu Leu Asp Ser Asp Gly Arg Thr Tyr 1 5 10 <210> 105 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR2 <400> 105 Leu Val Ser 1 <210> 106 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR3 <400> 106 Trp Gln Gly Thr His Phe Pro His Thr 1 5 <210> 107 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR1 <400> 107 Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn Tyr 1 5 10 <210> 108 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR2 <400> 108 Phe Ala Ser 1 <210> 109 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR3 <400> 109 Gln Gln His Tyr Ser Thr Pro Tyr Thr 1 5 <210> 110 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR1 <400> 110 Glu Ser Val Asp Asn Tyr Gly Ile Ser Phe 1 5 10 <210> 111 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR2 <400> 111 Ala Ala Ser 1 <210> 112 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR3 <400> 112 Gln Gln Ser Lys Glu Val Pro Phe Thr 1 5 <210> 113 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR1 <400> 113 Lys Asn Ile Ser Lys Tyr 1 5 <210> 114 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR2 <400> 114 Ser Gly Ser 1 <210> 115 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR3 <400> 115 Gln Gln His Asn Glu Tyr Pro Phe Thr 1 5 <210> 116 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR1 <400> 116 Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr 1 5 10 <210> 117 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR2 <400> 117 Trp Ala Ser 1 <210> 118 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR3 <400> 118 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 119 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR1 <400> 119 Gln Asp Ile Gly Ser Arg 1 5 <210> 120 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR2 <400> 120 Ala Thr Ser 1 <210> 121 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR3 <400> 121 Leu Gln Tyr Ala Ser Ser Pro Phe Thr 1 5 <210> 122 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR1 <400> 122 Asp His Ile Asp Asn Trp 1 5 <210> 123 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR2 <400> 123 Gly Ala Thr 1 <210> 124 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR3 <400> 124 Gln Gln Tyr Trp Ser Ser Pro Phe Thr 1 5 <210> 125 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR1 <400> 125 Gln Asp Ile Ser Asn Tyr 1 5 <210> 126 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR2 <400> 126 Tyr Thr Ser 1 <210> 127 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR3 <400> 127 Gln Gln Gly Asn Thr Leu Pro Trp Thr 1 5 <210> 128 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR1 <400> 128 Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr 1 5 10 <210> 129 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR2 <400> 129 Trp Ala Ser 1 <210> 130 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR3 <400> 130 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 131 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR1 <400> 131 Gln Asp Val Gly Thr Ala Val Ala 1 5 <210> 132 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR2 <400> 132 Trp Ala Ser 1 <210> 133 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR3 <400> 133 Gln Gln Tyr Ser Ser Tyr Pro Tyr Thr 1 5 <210> 134 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR1 <400> 134 Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr 1 5 10 <210> 135 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR2 <400> 135 Trp Ala Ser 1 <210> 136 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR3 <400> 136 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 137 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR1 <400> 137 Glu Asn Ile Tyr Ser Tyr 1 5 <210> 138 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR2 <400> 138 Asn Ala Lys 1 <210> 139 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR3 <400> 139 Gln His His Tyr Gly Thr Pro Phe Thr 1 5 <210> 140 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR1 <400> 140 Lys Ser Leu Leu His Ser Asn Asp Ile Thr Tyr 1 5 10 <210> 141 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR2 <400> 141 Arg Met Ser 1 <210> 142 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR3 <400> 142 Ala Gln Met Leu Glu Arg Pro Trp Thr 1 5 <210> 143 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR1 <400> 143 Ser Ser Val Ser Tyr 1 5 <210> 144 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR2 <400> 144 Leu Thr Ser 1 <210> 145 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR3 <400> 145 Gln Gln Trp Ser Ser Asn Pro Trp Thr 1 5 <210> 146 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR1 <400> 146 Gln Gly Ile Ser Asn Tyr 1 5 <210> 147 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR2 <400> 147 Tyr Thr Ser 1 <210> 148 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR3 <400> 148 Gln Gln Tyr Ser Lys Leu Pro Arg Thr 1 5 <210> 149 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR1 <400> 149 Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn Tyr 1 5 10 <210> 150 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR2 <400> 150 Phe Ala Ser 1 <210> 151 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR3 <400> 151 Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5 <210> 152 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR1 <400> 152 Glu Asn Ile Tyr Tyr Ser 1 5 <210> 153 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR2 <400> 153 Asn Ala Asp 1 <210> 154 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR3 <400> 154 Lys Gln Ala Tyr Asp Val Pro Leu Thr 1 5 <210> 155 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR1 <400> 155 Ser Ser Val Ser Tyr 1 5 <210> 156 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR2 <400> 156 Leu Thr Ser 1 <210> 157 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR3 <400> 157 Gln Gln Trp Ser Ser Asn Pro Pro Thr 1 5 <210> 158 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR1 <400> 158 Gln Asp Ile Asn Ser Tyr 1 5 <210> 159 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR2 <400> 159 Arg Ala Asn 1 <210> 160 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR3 <400> 160 Leu Gln Tyr Asp Glu Phe Pro Leu Thr 1 5 <210> 161 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR1 <400> 161 Gly Phe Ser Leu Ser Thr Phe Gly Met Gly Val Thr 1 5 10 <210> 162 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR2 <400> 162 His Ile Trp Trp Asp Asp Asp Met Tyr Tyr Asn Pro Ala Leu Lys Ser 1 5 10 15 <210> 163 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 37C7A HC CDR3 <400> 163 Ala Arg Ser Pro Ile Thr Thr Val Val Ala Asp Tyr 1 5 10 <210> 164 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR1 <400> 164 Gly Tyr Thr Phe Ser Ser Tyr Trp Ile Glu 1 5 10 <210> 165 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR2 <400> 165 Glu Ile Leu Pro Gly Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 166 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 38C16A HC CDR3 <400> 166 Ala Arg Gly Asp Tyr Phe Gly Ser Ser Tyr Arg Gly Pro Tyr 1 5 10 <210> 167 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR1 <400> 167 Gly Tyr Thr Phe Thr Ser Tyr Trp Met His 1 5 10 <210> 168 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR2 <400> 168 Glu Ile Asp Pro Ser Gly Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 169 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 39G8A HC CDR3 <400> 169 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr 1 5 10 <210> 170 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR1 <400> 170 Gly Tyr Thr Phe Ser Asn Tyr Trp Ile Glu 1 5 10 <210> 171 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR2 <400> 171 Glu Ile Leu Pro Gly Asn Val Ile Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 172 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 43E8A HC CDR3 <400> 172 Ala Arg Arg Gly Asp Asp Gly Tyr Leu Tyr Ala Met Asp Tyr 1 5 10 <210> 173 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR1 <400> 173 Gly Tyr Thr Phe Thr Asp Tyr Tyr Met His 1 5 10 <210> 174 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR2 <400> 174 Glu Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 175 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 46J3A HC CDR3 <400> 175 Ala Arg Tyr Trp Asp Tyr Tyr Gly Ser Thr Tyr Gly Tyr Phe Asp Val 1 5 10 15 <210> 176 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR1 <400> 176 Gly Tyr Thr Phe Thr Asp Tyr Asn Met His 1 5 10 <210> 177 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR2 <400> 177 Tyr Ile Asn Pro Asn Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 178 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 46O19A HC CDR3 <400> 178 Ala Arg Asp Tyr Phe Trp Tyr Phe Asp Val 1 5 10 <210> 179 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR1 <400> 179 Gly Phe Ser Leu Thr Ser Tyr Gly Val His 1 5 10 <210> 180 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR2 <400> 180 Val Ile Trp Ser Gly Gly Ser Thr Asp Tyr Asn Ala Ala Phe Ile Ser 1 5 10 15 Arg <210> 181 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 47H24A HC CDR3 <400> 181 Ala Ser Gln Tyr Val Ala Tyr 1 5 <210> 182 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR1 <400> 182 Gly Tyr Thr Phe Thr Ser Tyr Trp Met His 1 5 10 <210> 183 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR2 <400> 183 Asn Ile Asn Pro Ser Asn Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 184 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 49O5A HC CDR3 <400> 184 Ala Arg Ser Lys Ser Asn Tyr Gly Tyr Tyr Ala Met Asp Tyr 1 5 10 <210> 185 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR1 <400> 185 Gly Tyr Thr Phe Thr Ser Tyr Trp Met His 1 5 10 <210> 186 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR2 <400> 186 Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 187 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50B10A HC CDR3 <400> 187 Ala Arg Ser Glu Gly Arg Val Tyr Tyr Asp Tyr Phe Tyr Ala Met Asp 1 5 10 15 Tyr <210> 188 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR1 <400> 188 Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu 1 5 10 <210> 189 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR2 <400> 189 Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 190 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50H17A HC CDR3 <400> 190 Ala Arg Asp Tyr Tyr Trp Tyr Phe Asp Val 1 5 10 <210> 191 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR1 <400> 191 Ala Ile Arg Asp Thr Asn Tyr Trp Met Gln 1 5 10 <210> 192 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR2 <400> 192 Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Val <210> 193 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> 50E16A HC CDR3 <400> 193 Ala Thr Tyr Tyr Ser Asn Tyr Gly Gly Ala Met Asp Tyr 1 5 10 <210> 194 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR1 <400> 194 Gly Tyr Thr Phe Thr Asp Tyr Ser Met His 1 5 10 <210> 195 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR2 <400> 195 Trp Ile Asn Thr Glu Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 196 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 50F1A HC CDR3 <400> 196 Ala Arg Asp Ile Phe Trp Ala Met Asp Tyr 1 5 10 <210> 197 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR1 <400> 197 Gly Tyr Thr Phe Thr Ser Tyr Asn Met His 1 5 10 <210> 198 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR2 <400> 198 Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 199 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 54I14A HC CDR3 <400> 199 Ala Arg Tyr Asp Tyr Asp Ala Met Asp Tyr 1 5 10 <210> 200 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR1 <400> 200 Gly Tyr Ser Phe Thr Gly Tyr Asn Met Asn 1 5 10 <210> 201 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR2 <400> 201 Asn Ile Asn Pro Tyr Tyr Gly Ser Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 202 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 52K11A HC CDR3 <400> 202 Ala Gly Ser Ser Tyr Val Asp Tyr Ala Met Asp Tyr 1 5 10 <210> 203 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR1 <400> 203 Gly Tyr Thr Phe Thr Ser Tyr Val Met His 1 5 10 <210> 204 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR2 <400> 204 Tyr Ile Ile Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe Glu 1 5 10 15 Gly <210> 205 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 53H21A HC CDR3 <400> 205 Ala Arg Trp Gly Asn Trp Asp Tyr Phe Asp Tyr 1 5 10 <210> 206 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR1 <400> 206 Gly Tyr Thr Phe Thr Ser Tyr Trp Ile Asn 1 5 10 <210> 207 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR2 <400> 207 Asn Ile Tyr Ala Gly Ser Ser Ser Ser Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 208 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> 53I12A HC CDR3 <400> 208 Ala Arg Ser Gly His Gly Tyr Asp Gly Phe Ala Tyr 1 5 10 <210> 209 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR1 <400> 209 Gly Tyr Ala Phe Ser Ser Tyr Trp Met Asn 1 5 10 <210> 210 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR2 <400> 210 Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Lys Phe Lys 1 5 10 15 Gly <210> 211 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 60A9A HC CDR3 <400> 211 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr 1 5 10 <210> 212 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR1 <400> 212 Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu 1 5 10 <210> 213 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR2 <400> 213 Leu Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Ile Glu Lys Phe Lys 1 5 10 15 Gly <210> 214 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> 39J9A HC CDR3 <400> 214 Ala Arg Arg Gly Asp Tyr Tyr Gly Asn Pro Phe Asp Tyr 1 5 10 <210> 215 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR1 <400> 215 Gly Tyr Thr Phe Thr Asn Tyr Trp Met Gln 1 5 10 <210> 216 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR2 <400> 216 Glu Ile Asp Pro Ser Asp Asn Tyr Thr His Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 217 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 49H8A HC CDR3 <400> 217 Ala Arg Gly Tyr Tyr Gly Tyr Ser Pro Ser Trp Phe Ala Tyr 1 5 10 <210> 218 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR1 <400> 218 Gly Tyr Ser Phe Thr Gly Tyr Tyr Met His 1 5 10 <210> 219 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR2 <400> 219 Arg Ile Asn Pro Tyr Ser Gly Ala Thr Asn Ser Asn Gln Asn Phe Lys 1 5 10 15 Asp <210> 220 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 51B10A HC CDR3 <400> 220 Ala Arg Ser Tyr Tyr Gly Ala Met Asp Tyr 1 5 10 <210> 221 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR1 <400> 221 Arg Ser Ser Gln Asn Leu Val His Ser Tyr Gly Asn Thr Tyr Leu His 1 5 10 15 <210> 222 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR2 <400> 222 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 223 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 37C7A LC CDR3 <400> 223 Ser Gln Asn Thr His Val Pro Trp Thr 1 5 <210> 224 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR1 <400> 224 Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Arg Thr Tyr Leu Asn 1 5 10 15 <210> 225 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR2 <400> 225 Leu Val Ser Lys Leu Asp Ser 1 5 <210> 226 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 38C16A LC CDR3 <400> 226 Trp Gln Gly Thr His Phe Pro His Thr 1 5 <210> 227 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR1 <400> 227 Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn Tyr Leu 1 5 10 15 Ala <210> 228 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR2 <400> 228 Phe Ala Ser Thr Arg Glu Ser 1 5 <210> 229 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 39G8A LC CDR3 <400> 229 Gln Gln His Tyr Ser Thr Pro Tyr Thr 1 5 <210> 230 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR1 <400> 230 Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Ile Ser Phe Met Asn 1 5 10 15 <210> 231 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR2 <400> 231 Ala Ala Ser Asn Gln Gly Ser 1 5 <210> 232 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 43E8A LC CDR3 <400> 232 Gln Gln Ser Lys Glu Val Pro Phe Thr 1 5 <210> 233 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR1 <400> 233 Arg Ala Ser Lys Asn Ile Ser Lys Tyr Leu Ala 1 5 10 <210> 234 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR2 <400> 234 Ser Gly Ser Thr Leu Gln Ser 1 5 <210> 235 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 46J3A LC CDR3 <400> 235 Gln Gln His Asn Glu Tyr Pro Phe Thr 1 5 <210> 236 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR1 <400> 236 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu 1 5 10 15 Thr <210> 237 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR2 <400> 237 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 238 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 46O19A LC CDR3 <400> 238 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 239 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR1 <400> 239 Arg Ala Ser Gln Asp Ile Gly Ser Arg Leu Thr 1 5 10 <210> 240 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR2 <400> 240 Ala Thr Ser Ser Leu Asp Ser 1 5 <210> 241 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 47H24A LC CDR3 <400> 241 Leu Gln Tyr Ala Ser Ser Pro Phe Thr 1 5 <210> 242 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR1 <400> 242 Glu Ala Ser Asp His Ile Asp Asn Trp Leu Ala 1 5 10 <210> 243 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR2 <400> 243 Gly Ala Thr Ser Leu Glu Thr 1 5 <210> 244 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 49O5A LC CDR3 <400> 244 Gln Gln Tyr Trp Ser Ser Pro Phe Thr 1 5 <210> 245 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR1 <400> 245 Arg Ala Ser Gln Asp Ile Ser Asn Tyr Leu Asn 1 5 10 <210> 246 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR2 <400> 246 Tyr Thr Ser Arg Leu His Ser 1 5 <210> 247 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50B10A LC CDR3 <400> 247 Gln Gln Gly Asn Thr Leu Pro Trp Thr 1 5 <210> 248 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR1 <400> 248 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu 1 5 10 15 Thr <210> 249 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR2 <400> 249 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 250 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50H17A LC CDR3 <400> 250 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 251 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR1 <400> 251 Lys Ala Ser Gln Asp Val Gly Thr Ala Val Ala 1 5 10 <210> 252 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR2 <400> 252 Trp Ala Ser Thr Arg His Thr 1 5 <210> 253 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50E16A LC CDR3 <400> 253 Gln Gln Tyr Ser Ser Tyr Pro Tyr Thr 1 5 <210> 254 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR1 <400> 254 Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu 1 5 10 15 Thr <210> 255 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR2 <400> 255 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 256 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 50F1A LC CDR3 <400> 256 Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 1 5 <210> 257 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR1 <400> 257 Arg Ala Ser Glu Asn Ile Tyr Ser Tyr Phe Ala 1 5 10 <210> 258 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR2 <400> 258 Asn Ala Lys Thr Leu Ala Glu 1 5 <210> 259 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 54I14A LC CDR3 <400> 259 Gln His His Tyr Gly Thr Pro Phe Thr 1 5 <210> 260 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR1 <400> 260 Ser Ser Asn Lys Ser Leu Leu His Ser Asn Asp Ile Thr Tyr Leu Tyr 1 5 10 15 <210> 261 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR2 <400> 261 Arg Met Ser Asn Leu Ala Ser 1 5 <210> 262 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 52K11A LC CDR3 <400> 262 Ala Gln Met Leu Glu Arg Pro Trp Thr 1 5 <210> 263 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR1 <400> 263 Ser Ala Ser Ser Ser Val Ser Tyr Met Tyr 1 5 10 <210> 264 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR2 <400> 264 Leu Thr Ser Asn Leu Ala Ser 1 5 <210> 265 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 53H21A LC CDR3 <400> 265 Gln Gln Trp Ser Ser Asn Pro Trp Thr 1 5 <210> 266 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR1 <400> 266 Ser Ala Ser Gln Gly Ile Ser Asn Tyr Leu Asn 1 5 10 <210> 267 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR2 <400> 267 Tyr Thr Ser Ser Leu His Ser 1 5 <210> 268 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 53I12A LC CDR3 <400> 268 Gln Gln Tyr Ser Lys Leu Pro Arg Thr 1 5 <210> 269 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR1 <400> 269 Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn Tyr Leu 1 5 10 15 Ala <210> 270 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR2 <400> 270 Phe Ala Ser Thr Arg Asp Ser 1 5 <210> 271 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 60A9A LC CDR3 <400> 271 Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5 <210> 272 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR1 <400> 272 Arg Ala Ser Glu Asn Ile Tyr Tyr Ser Leu Ala 1 5 10 <210> 273 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR2 <400> 273 Asn Ala Asp Thr Leu Glu Asp 1 5 <210> 274 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 39J9A LC CDR3 <400> 274 Lys Gln Ala Tyr Asp Val Pro Leu Thr 1 5 <210> 275 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR1 <400> 275 Ser Ala Ser Ser Ser Val Ser Tyr Met Tyr 1 5 10 <210> 276 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR2 <400> 276 Read Thr Ser Asn Read Wing Ser 1 5 <210> 277 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 49H8A LC CDR3 <400> 277 Gln Gln Trp Ser Ser Asn Pro Pro Thr 1 5 <210> 278 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR1 <400> 278 Lys Ala Ser Gln Asp Ile Asn Ser Tyr Leu Ser 1 5 10 <210> 279 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR2 <400> 279 Arg Ala Asn Arg Leu Val Asp 1 5 <210> 280 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 51B10A LC CDR3 <400> 280 Leu Gln Tyr Asp Glu Phe Pro Leu Thr 1 5 <210> 281 <211> 574 <212> PRT <213> Homo sapiens <400> 281 Met Cys Pro Arg Ala Ala Arg Ala Pro Ala Thr Leu Leu Leu Ala Leu 1 5 10 15 Gly Ala Val Leu Trp Pro Ala Ala Gly Ala Trp Glu Leu Thr Ile Leu 20 25 30 His Thr Asn Asp Val His Ser Arg Leu Glu Gln Thr Ser Glu Asp Ser 35 40 45 Ser Lys Cys Val Asn Ala Ser Arg Cys Met Gly Gly Val Ala Arg Leu 50 55 60 Phe Thr Lys Val Gln Gln Ile Arg Arg Ala Glu Pro Asn Val Leu Leu 65 70 75 80 Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr Ile Trp Phe Thr Val Tyr 85 90 95 Lys Gly Ala Glu Val Ala His Phe Met Asn Ala Leu Arg Tyr Asp Ala 100 105 110 Met Ala Leu Gly Asn His Glu Phe Asp Asn Gly Val Glu Gly Leu Ile 115 120 125 Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro Ile Leu Ser Ala Asn Ile 130 135 140 Lys Ala Lys Gly Pro Leu Ala Ser Gln Ile Ser Gly Leu Tyr Leu Pro 145 150 155 160 Tyr Lys Val Leu Pro Val Gly Asp Glu Val Val Gly Ile Val Gly Tyr 165 170 175 Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn Pro Gly Thr Asn Leu Val 180 185 190 Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro Glu Val Asp Lys Leu Lys 195 200 205 Thr Leu Asn Val Asn Lys Ile Ile Ala Leu Gly His Ser Gly Phe Glu 210 215 220 Met Asp Lys Leu Ile Ala Gln Lys Val Arg Gly Val Asp Val Val Val 225 230 235 240 Gly Gly His Ser Asn Thr Phe Leu Tyr Thr Gly Asn Pro Pro Ser Lys 245 250 255 Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile Val Thr Ser Asp Asp Gly 260 265 270 Arg Lys Val Pro Val Val Gln Ala Tyr Ala Phe Gly Lys Tyr Leu Gly 275 280 285 Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly Asn Val Ile Ser Ser His 290 295 300 Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile Pro Glu Asp Pro Ser Ile 305 310 315 320 Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys Leu Asp Asn Tyr Ser Thr 325 330 335 Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu Asp Gly Ser Ser Gln Ser 340 345 350 Cys Arg Phe Arg Glu Cys Asn Met Gly Asn Leu Ile Cys Asp Ala Met 355 360 365 Ile Asn Asn Asn Leu Arg His Thr Asp Glu Met Phe Trp Asn His Val 370 375 380 Ser Met Cys Ile Leu Asn Gly Gly Gly Ile Arg Ser Pro Ile Asp Glu 385 390 395 400 Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn Leu Ala Ala Val Leu Pro 405 410 415 Phe Gly Gly Thr Phe Asp Leu Val Gln Leu Lys Gly Ser Thr Leu Lys 420 425 430 Lys Ala Phe Glu His Ser Val His Arg Tyr Gly Gln Ser Thr Gly Glu 435 440 445 Phe Leu Gln Val Gly Gly Ile His Val Val Tyr Asp Leu Ser Arg Lys 450 455 460 Pro Gly Asp Arg Val Val Lys Leu Asp Val Leu Cys Thr Lys Cys Arg 465 470 475 480 Val Pro Ser Tyr Asp Pro Leu Lys Met Asp Glu Val Tyr Lys Val Ile 485 490 495 Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp Gly Phe Gln Met Ile Lys 500 505 510 Asp Glu Leu Leu Arg His Asp Ser Gly Asp Gln Asp Ile Asn Val Val 515 520 525 Ser Thr Tyr Ile Ser Lys Met Lys Val Ile Tyr Pro Ala Val Glu Gly 530 535 540 Arg Ile Lys Phe Ser Thr Gly Ser His Cys His Gly Ser Phe Ser Leu 545 550 555 560 Ile Phe Leu Ser Leu Trp Ala Val Ile Phe Val Leu Tyr Gln 565 570 <210> 282 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H1 Heavy Chain Variable Region <400> 282 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gln 1 5 10 15 Ser Leu Lys Leu Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Asn Pro Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 283 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H2 Heavy Chain Variable Region <400> 283 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 Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Asn Pro Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 284 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H3 Heavy Chain Variable Region <400> 284 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gln 1 5 10 15 Ser Leu Lys Leu Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Asn Pro Ser Val 50 55 60 Lys Gly Arg Ala Thr Leu Ser Ala Asp Lys Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 285 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H4 Heavy Chain Variable Region <400> 285 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gln 1 5 10 15 Ser Leu Lys Leu Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Ser Val 50 55 60 Lys Gly Arg Ala Thr Leu Ser Ala Asp Lys Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 286 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H5 Heavy Chain Variable Region <400> 286 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gln 1 5 10 15 Ser Leu Lys Leu Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Lys Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Ser Ala Asp Lys Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 287 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 39G8-H1 Heavy Chain Variable Region <400> 287 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 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asp Pro Ser Gly Gly Tyr Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Ser Thr Leu Thr Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 288 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 39G8-H2 Heavy Chain Variable Region <400> 288 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 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asp Pro Ser Gly Gly Tyr Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Ser Thr Leu Thr Val Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 289 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 39G8-H3 Heavy Chain Variable Region <400> 289 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 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asp Pro Ser Gly Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Ser Thr Leu Thr Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Tyr Tyr Gly Ser Ser Gly Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 290 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9-L1 Light Chain Variable Region <400> 290 Asp Ile Gln Met Thr Gln Ser Pro Ser Leu Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Asp 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 Glu Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 291 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9-L2 Light Chain Variable Region <400> 291 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Asp 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 Glu Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 292 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9-L3 Light Chain Variable Region <400> 292 Asp Ile Gln Met Thr Gln Ser Pro Ser Leu Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Asp 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 Glu Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 293 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9-L4 Light Chain Variable Region <400> 293 Asp Ile Val Met Thr Gln Ser Pro Ser Leu Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Asp 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 Glu Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile 100 105 110 Lys <210> 294 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 39G8-L3 Light Chain Variable Region <400> 294 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Val Tyr Phe 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 Phe Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105 110 Lys <210> 295 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H6 Heavy Chain Variable Region <400> 295 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Arg Pro Gly Ser 1 5 10 15 Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ala Pro Arg Phe 50 55 60 Gln Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Leu Glu Leu Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 296 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H7 Heavy Chain Variable Region <400> 296 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ala Pro Lys Phe 50 55 60 Gln Gly Arg Ala Thr Leu 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 Phe Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 297 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H8 Heavy Chain Variable Region <400> 297 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Lys Phe 50 55 60 Gln Gly Arg Ala Thr Leu 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 Phe Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 298 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 60A9-H9 Heavy Chain Variable Region <400> 298 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Arg Pro Gly Ser 1 5 10 15 Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Gln Ile Tyr Pro Gly Asp Gly Asp Thr Tyr Tyr Ser Gly Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Ser Ala Asp Lys Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Asn Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ile Tyr Tyr Gly Asn Tyr Val Phe Thr Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 299 <211> 113 <212> PRT <213> Artificial Sequence <220> <223> 60A9-L5 Light Chain Variable Region <400> 299 Glu Ile Val Met Thr Gln Ser Pro Gly Thr Gln Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Arg Pro Gly Gln 35 40 45 Ala Pro Arg Leu Leu Val Tyr Phe Ala Ser Thr Arg Asp Ser Gly Val 50 55 60 Ala Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln 85 90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Val 100 105 110 Lys

Claims

1. d. SEQ ID NOs: 47, 48, 49, 107, 108, and 109, respectively, or SEQ ID NOs: 167, 168, 169, 227, 228, and 229, respectively; 5. An isolated monoclonal antibody or antigen-binding fragment thereof, which specifically binds to CD73, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

2. 2. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which specifically binds to human CD73.

3. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 or 2, comprising a heavy chain variable region having a polypeptide sequence that is at least 95% identical to SEQ ID NO: 5, 287, 288, or 289, or a light chain variable region having a polypeptide sequence that is at least 95% identical to SEQ ID NO: 6 or 294.

4. e. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:5, and a light chain variable region having the polypeptide sequence of SEQ ID NO:6; nn. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:287, and a light chain variable region having the polypeptide sequence of SEQ ID NO:294; oo. a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 288, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 294; or pp. a heavy chain variable region having the polypeptide sequence of SEQ ID NO:289, and a light chain variable region having the polypeptide sequence of SEQ ID NO:294; The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, comprising:

5. The isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, which is capable of inhibiting the enzymatic activity of soluble and / or cell surface CD73 and / or activating T cells.

6. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 5, which is chimeric or humanized.

7. A bispecific antibody comprising the monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 6.

8. An isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody according to any one of claims 1 to 7.

9. A vector comprising the isolated nucleic acid of claim 8.

10. 10. A host cell comprising the isolated nucleic acid of claim 8 or the vector of claim 9.

11. A pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody according to any one of claims 1 to 7, and a pharma- ceutically acceptable carrier.

12. The pharmaceutical composition of claim 11 for inhibiting the nucleotidase activity of CD73 or for treating cancer in a subject in need thereof.

13. 8. A method for producing the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody according to any one of claims 1 to 7, comprising the steps of culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody, under conditions for producing the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody, and recovering the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody from the cell or culture.

14. 8. A method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody according to any one of claims 1 to 7, comprising the step of combining the monoclonal antibody or antigen-binding fragment thereof, or bispecific antibody with a pharma- ceutically acceptable carrier to obtain a pharmaceutical composition.

15. 1. A method for determining the level of CD73 or determining the ecto-5'-nucleotidase activity of CD73 in a subject, comprising: a. contacting a sample from the subject with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 6; and b. determining the level of CD73 or determining the ecto-5'-nucleotidase activity of CD73 in said subject. The method comprising:

16. The method of claim 15, wherein the sample is a tissue sample or a blood sample.

17. The method of claim 16, wherein the tissue sample is a cancer tissue sample.

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