EA039293B1 - Anti-lag3 antibodies and antigen-binding fragments - Google Patents

Abstract

The invention relates to antibodies and antigen-binding fragments thereof that specifically bind to human LAG3 as well as fragments thereof. The invention relates to a polynucleotide encoding light and heavy chains of the claimed antibodies, to an expression vector comprising such polynucleotide, to pharmaceutical compositions comprising such antibodies as a single active agent or in association with another active agent.

Description

(57) The invention relates to antibodies and their antigen-binding fragments that specifically bind to human LAG3, as well as their variants. The invention relates to a polynucleotide encoding the light and heavy chains of the claimed antibodies, to an expression vector containing such a polynucleotide, to pharmaceutical compositions containing such antibodies as the sole active ingredient or in combination with another active ingredient.

039293 Bl

This patent application claims priority to US Provisional Application No. 62/039081, filed August 19, 2014; and US Provisional Application No. 62/171319, filed June 5, 2015; all of which are incorporated herein by reference in their entirety.

The invention contains a sequence listing which is provided in electronic form in ASCII format and is thus incorporated herein by reference in its entirety. The specified ASCII copy, created on June 29, 2015, is named 23791_seqlist and is 967 kilobytes in size.

Field of invention

The present invention relates to antibodies against LAG3, as well as the use of the antibodies of the present invention for the treatment of diseases such as cancer and infection.

The state of the art to which the invention relates

LAG3 (CD223) is a cell surface molecule expressed on activated T cells (Huard et al. Immunogenetics 39:213-217, 1994), NK cells (Triebel et al. J Exp Med 171:1393-1405, 1990) , B cells (Kisielow et al. Eur J Immunol 35:2081-2088, 2005) and plasmacytoid dendritic cells (Workman et al. J Immunol 182:1885-1891, 2009), which play an important role in the function of these lymphocyte subgroups. In addition, the interaction between LAG3 and its main ligand, MHC class II, is believed to be involved in modulating dendritic cell function (Andreae et al. J Immunol 168:3874-3880, 2002). The role of LAG-3 in CD8 T cell depletion has been documented in recent preclinical studies (Blackburn et al. Nat Immunol 10:29-37, 2009).

As with chronic viral infection, tumor antigen-specific CD4+ and CD8+ T cells exhibit reduced effector function and a depleted phenotype characterized by reduced production of pro-inflammatory cytokines and reduced responsiveness to antigen restimulation. This is mediated by extrinsic cellular mechanisms such as regulatory T cells (Treg) and intrinsic cellular mechanisms such as inhibitory molecules that are activated on depleted tumor infiltrating lymphocytes (TILs). These inhibitory mechanisms are a formidable barrier to effective antitumor immunity.

LAG is expressed on non-immunogenic TILs, indicating that they contribute to tumor-mediated immune suppression. Inhibition of LAG3 may result in enhanced activation of antigen-specific T cells, from which therapeutic benefits may be derived. There is a need in the art for highly effective therapeutic antibodies that antagonize LAG3 activity that can be used to achieve a robust immune response to tumors.

The essence of the invention

The present invention relates to an antibody or antigen-binding fragment thereof (e.g., antibody, antigen-binding fragment, monoclonal antibodies, polyclonal antibodies, multispecific antibody, humanized antibody, isolated antibody or antigen-binding fragment thereof, humanized antagonist antibody, fully human antibody, chimeric antibody, and camelized single-domain antibody ) that specifically bind LAG3 (e.g., human and/or Macaqa fascicularis, e.g., SEQ ID NO: 443 or 445) containing: an immunoglobulin heavy chain variable region having at least 78.99% amino acid sequence identity with amino acids 1- 119 SEQ ID NO: 106; and/or an immunoglobulin light chain variable region having at least 78.38% amino acid sequence identity with amino acids 1-111 of SEQ ID NO: 224.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention comprises an immunoglobulin light and heavy chain (e.g., heavy and light chain variable domains or heavy and light chain CDRs) Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7 , Ab8 or Ab9 as described herein.

The present invention relates to an antibody or antigen-binding fragment thereof that specifically binds LAG3 (e.g., human and cynomolgus monkey LAG3) containing (a) CDR1, CDR2 and CDR3 of an immunoglobulin VL chain domain, which contains the amino acid sequence specified in

SEQ ID NO: 7, 17, 27, 37, 57, 59, 61, 63, 65, 101, 126, 130, 132, 136,

138 208 210 224 226 228 230 232 241 257 259 261 263 351 369 371 373 375 401 403 405 426 427 450-453 or 459-461;

and/or (b) CDR1, CDR2 and CDR3 of the V _n- domain chain of an immunoglobulin that contains the amino acid sequence specified in

- 1 039293

SEQ ID NO: 2, 12, 22, 32, 45, 47, 49, 51, 53, 55, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 128, 134, 140, 142, 144, 146, 148, 150, 152, 154,

156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178,180,

182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204,206,

212, 214, 216, 218, 220, 222, 234, 235, 237, 239, 243, 245,247,

249, 251, 253, 255, 265, 267, 269, 271, 273, 275, 277, 279.281,

283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307,

309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331,333,

335, 337, 339, 341, 343, 345, 347, 349, 353, 355, 357, 359,361,

363, 365, 367, 377, 379, 381, 383, 385, 387, 389, 391, 393,395,

397, 399, 406-419, 434-442, 448, 449, 462, 463 or 464.

The present invention also relates to an antibody or antigen-binding fragment thereof that specifically binds LAG3 (e.g., human or cynomolgus monkey) containing (1) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQID NO: 38) ; CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence: dynvd (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence: DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (2) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQid no: 39); and CDR-L3, which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: Loy chain containing CDR-H1, which contains dynvd (seq id NO: 33); CDR-H2, which contains

0); and/or variable domain heavy amino acid sequence: amino acid sequence:

dinpnsggtiyaqkfqe (seq id NO: 456); and CDR-H3 which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35) _; or (3) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence: dynvd (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence: DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (4) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 which contains the amino acid sequence: DINPNQGGTIYAQKFQE (SEQ ID NO: 455). and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35) - or (5) light chain variable domain containing CDR-L1, which contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence: gasnles (seq id NO: 39); and CDR-L3 which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO:33); CDR-H2, which contains the amino acid sequence: DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3 which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35).

In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds LAG3 (e.g., human or cynomolgus LAG3) contains the CDRs of various light and/or heavy chain variable regions and possesses at least 90%

- 2 039293 common amino acid sequence identity with the variable region, i.e. chain variability occurs outside of CDRs, for example, (1) an immunoglobulin light chain containing CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence shown in SEQ ID NO: 126 or amino acids 21-131 thereof and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 106 or its amino acids 1-119, and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 106, or its amino acids 1-119; (2) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 108, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 108, or its amino acids 1-119; (3) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 110, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 110, or its amino acids 1-119; (4) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 112, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 112, or its amino acids 1-119; (5) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDRH1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 114, or amino acids 1-119 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 114, or its amino acids 1-119; (6) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 116, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 116, or its amino acids 1-119; (7) an immunoglobulin light chain comprising CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 118, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 118, or its amino acids 1-119; (8) an immunoglobulin light chain containing CDR-L1, CDRL2 and CDR-L3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain comprising CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin comprising the amino acid sequence of SEQ ID NO: 120, or amino acids 1-119 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 120, or its amino acids 1-119; and/or (9)

- 3 039293 an immunoglobulin light chain containing CDR-L1, CDR-L2 and CDR-L3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof, and having at least 90% amino acid sequence identity with the amino acid sequence, specified in SEQ ID NO: 126, or amino acids 21-131 thereof, and/or an immunoglobulin heavy chain containing CDR-H1, CDR-H2 and CDR-H3 of an immunoglobulin containing the amino acid sequence of SEQ ID NO: 122, or amino acids 1-119 thereof , and having at least 90% amino acid sequence identity with the amino acid sequence specified in SEQ ID NO: 122, or its amino acids 1-119. In one embodiment, the antibody or antigen-binding fragment thereof comprises distinct mature immunoglobulin light chain variable regions and/or mature immunoglobulin heavy chain variable regions and shares at least 90% amino acid sequence identity with the full length immunoglobulin variable region (including the signal sequence), i.e. e. variability occurs outside the sequences of mature immunoglobulin chains.

The present invention relates to an antibody or antigen-binding fragment specifically binding to LAG3 (for example, human or cynomolgus monkey LAG3) that contains a mature or full-length V _L domain or an immunoglobulin light chain.

SEQ ID NO: 7, 17, 27, 37, 57, 59, 61, 63, 65, 101,

126, 130, 132, 136, 138, 208, 210, 224, 226, 228, 230, 232, 241,

257, 259, 261, 263, 351, 369, 371, 373, 375, 401, 403, 405, 426, 427, 451-453 or 459-461;

and/or mature or full length VH domain or immunoglobulin heavy chain SEQ ID NOs: 2, 12, 22,

32, 45, 47, 49, 51, 53, 55, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 128, 134, 140, 142, 144, 146, 148,150,

152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174,176,

178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200.202,

204, 206, 212, 214, 216, 218, 220, 222, 234, 235, 237, 239.243,

245, 247, 249, 251, 253, 255, 265, 267, 269, 271, 273, 275,277,

279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301,303,

305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327,329,

331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 353, 355,357,

359, 361, 363, 365, 367, 377, 379, 381, 383, 385, 387, 389,391,

393, 395, 397, 399, 406-419, 434-442, 448, 449, 462, 463 or 464.

In one embodiment, the antibody or antigen-binding fragment thereof comprises: (1) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ lot sequence: GASNLES (SEQ id lot sequence: QQSTEDPRT (loose chain seq containing CDR-H1, which is DYNVD (SEQ ID NO:33); CDR-H2, which

ID NO: 38); CDR-L2, which contains amino acids NO: 39); and CDR-L3, which contains an amino acid ^{ID N0:} 40); and/or variable domain heavy contains an amino acid sequence: contains an amino acid sequence:

dinpnnggtiyaqkfqe (seq id NO: 458); and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (2) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence: DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (3) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence: QQSTEDPRT (SEQ ID no: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence:

- 4 039293

DINPNDGGTIYAQKFQE (SEQ ID NO: 457); _and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (4) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ id no: 39); and CDR-L3 which contains the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence: DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and CDR-H3, which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (5) a light chain variable domain containing a CDR-L1 that contains the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 which contains the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence: QQSTEDPRT (SEQ id no: 40); and/or a heavy chain variable domain containing a CDR-H1 which contains the amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence: DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3 which contains the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35).

In one embodiment, the antibody or fragment is glycosylated with yeast engineered N-linked glycans or CHO N-linked glycans. Optionally, any of the anti-LAG3 antibodies or antigen-binding fragments thereof of the invention has one or more of the following properties: inhibits LAG3 binding to class II MHC molecules; competes with MHC class II molecules for LAG3 binding; connects an additional loop of LAG3; binds LAG3 with K _D from about 10 ^-9 M to about 2x10' ¹² M; binds to native LAG3 on the surface of activated CD4+ and/or CD8+ T cells; binds to human and/or cynomolgus monkey LAG3; inhibits LAG3 homodimerization; stimulates the production of IL-2 by antigen-specific T cells; carries out labeling of tissue of the tonsil; does not label brain, heart, kidney, liver, lung, pancreas and/or pituitary tissue; binds to human LAG3 by contact with residues QEGAPAQL (amino acids 35-42 SEQ ID NO: 443) and RPARRADAGEYRAAVH (amino acids 137-152 SEQ ID NO: 443) and optionally residues DERGRQRGDFSLW (amino acids 123-135 SEQ ID NO: 443) LAG3; or residues SPTIPLQDL (amino acids 45-53 SEQ ID NO: 443) and optionally DERGRQRGDFSL (amino acids 123-134 SEQ ID NO: 443) LAG3; or residues HPLAPGPHPAAPSSWGPRPRRYTVL (amino acids 78-102 SEQ ID NO: 443) LAG3; and/or by protecting the hydrogen atoms on the amide backbone of such residues from being replaced by deuterium. The present invention also relates to any such antibody or fragment in a pharmaceutically acceptable carrier or diluent. In one embodiment, the anti-LAG3 antibody or fragment is immobilized on a solid support. In one embodiment, the LAG3 antibody, or antigen-binding fragment thereof, is Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, or Ab9 (as defined herein).

The present invention also relates to a complex containing an anti-LAG3 antibody or a fragment described in the present description (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/ or Ab9), in complex with LAG3 (eg, human or cynomolgus monkey) or a fragment thereof, or with a secondary antibody (eg, a detectably labeled secondary antibody) that specifically binds to an anti-LAG3 antibody or fragment thereof. In one embodiment, the antibody or fragment thereof is present in vitro (eg, immobilized on a solid support) or is in the body of an individual. In one embodiment of the invention, LAG3 is present in vitro (eg, immobilized on a solid support), or is on the surface of a cell, or is in the body of an individual.

In addition, the present invention relates to a composition containing a plurality of anti-LAG3 antibodies or antigen-binding fragments of the present invention, which are isolated monoclonal antibodies or antigen-binding fragments thereof, for example, which are humanized, for example, humanized antagonistic antibodies and antigen-binding fragments thereof.

The present invention also relates to a composition comprising anti-LAG3 antibodies or antigen-binding fragments thereof, as described herein, associated with an additional drug (for example, a monoclonal antibody or an antigen-binding fragment, or an organic small molecular weight compound), such as an immunomodulatory receptor inhibitor, an antiemetic agent, MTOR inhibitor (mammalian target of rapamycin), cytotoxic agent, platinum-based agent, EGFR (epidermal growth factor receptor) inhibitor, VEGF (vascular epidermal growth factor) inhibitor, microtubule stabilizer, taxane,

- 5 039293 CD20 inhibitor, CD52 inhibitor, CD30 inhibitor, RANK inhibitor (nuclear factor kappa B receptor activator), RANKL inhibitor (nuclear factor kappa B receptor activator ligand), ERK inhibitor, MAP kinase inhibitor, AKT inhibitor, MEK inhibitor , PI3K inhibitor, HER1 inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, Bcl2 inhibitor, CD22 inhibitor, CD79b inhibitor, ErbB2 inhibitor, and/or farnesyl protein transferase inhibitor. In one embodiment, the additional drug is an anti-PD1 antibody or antigen-binding fragment thereof, pembrolizumab, nivolumab, CT-011, anti-CTLA4 antibody or antigen-binding fragment thereof, anti-TIM3 antibody or antigen-binding fragment thereof, anti-CS1 antibody or antigen-binding fragment thereof, elotuzumab, anti-KIR2DL1/2/3 antibody or antigen-binding fragment thereof, lirilumab, anti-CD137 antibody or antigen-binding fragment thereof, urelumab, anti-GITR antibody or antigen-binding fragment thereof, TRX518, anti-PD-L1 antibody or antigen-binding fragment thereof, BMS-936559, MSB0010718C, MPDL3280A, anti-PD-L2 antibody or antigen binding fragment thereof, anti ILT1 antibody or antigen binding fragment thereof, anti ILT2 antibody or antigen binding fragment thereof, anti ILT3 antibody or antigen binding fragment thereof, anti ILT4 antibody or antigen binding fragment thereof, anti ILT5 antibody or e antigen-binding fragment, anti-ILT6 antibody or antigen-binding fragment thereof, anti-ILT7 antibody or antigen-binding fragment thereof, anti-ILT8 antibody or antigen-binding fragment thereof, anti-CD40 antibody or antigen-binding fragment thereof, anti-OX40 antibody or antigen-binding fragment thereof, anti-CD137 antibody or antigen-binding fragment thereof fragment, anti-KIR2DL1 antibody or antigen-binding fragment thereof, anti-KIR2DL2/3 antibody or antigen-binding fragment thereof, anti-KIR2DL4 antibody or antigen-binding fragment thereof, anti-KIR2DL5A antibody or antigen-binding fragment thereof, anti-KIR2DL5B antibody or antigen-binding fragment thereof, anti-KIR3DL1 antibody or antigen-binding fragment thereof fragment, anti-KIR3DL2 antibody or antigen-binding fragment thereof, anti-KIR3DL3 antibody or antigen-binding fragment thereof, anti-NKG2A antibody or antigen-binding fragment thereof, anti-NKG2C antibody or antigen-binding fragment thereof, and/or ant an anti-NKG2E body or antigen-binding fragment thereof, or any low molecular weight organic inhibitor of such targets; IL-10, anti-IL10 agent, anti-TSLP agent and/or pegylated IL-10. In one embodiment, the additional drug is 13-cis-retinoic acid, 3-[5(methylsulfonylpiperadinemethyl)indolyl]quinolone, 4-hydroxytamoxifen, 5-deoxyuridine, 5'deoxy-5-fluorouridine, 5-fluorouracil, 6- mercaptopurine, 7-hydroxystaurosporine, A-443654, abiraterone acetate, abraxane, ABT-578, acolbifen, ADS-100380, aflibercept, ALT-110, altretamine, amifostine, aminoglutethimide, amrubicin, amsacrine, anagrelide, anastrozole, angiostatin, AP-235 ARQ-197, arzoxifene, AS-252424, AS-605240, asparaginase, AT-9263, ATI3387, atrasentan, axitinib, AZD1152, Bacillus Calmette-Guerin (BCG) vaccine, batabulin, BC-210, BGJ398, bezodutox, bevacizumab Bicalutamide Bio111 BIO140 BKM120 Bleomycin , CG-781, chlamydocin, chlorambucil, chlorotoxin, cilengitide, cymiti din, cisplatin, cladribine, clodronate, cobimetnib, COL-3, CP-724714, cyclophosphamide, cyproterone, cyproterone acetate, cytarabine, cytosine arabinoside, dabrafenib, dacarbazine, dacinostat, dactinomycin, daltouzumab, danusertib, dasatinib, daunorubicin, decatinib, deguelin, deoxycoformycin, depsipeptide, diarylpropionitrile, diethylstilbestrol, diftitox, DNE03, docetaxel, dovitinib, doxorubicin, droloxifene, etothecarin, 90 yttrium-labeled edotreotide, edotreotide, EKB-569, EMD121974, encorafenib, endostatin, enzalutamide, E-rubiscin, RA, Viticine 923, erbitux, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, ficlatuzumab, finasteride, flavopiridol, floxuridine, fludarabine, fludrocortisone, fluoxymesterone, flutamide, FOLFOX regimen, fulvestrant, galeterone, ganetespib, gefitinib, himatelinib, gematelinib, gemcitabine acetate, gossypol, GSK461364, GSK690693, HMR-3339, hydroxyprogesterone caproate, hydroxyurea, IC87114, idarubicin, idoxifen, ifos famide, IM862, imatinib, IMC-1C11, INCB24360, INC280, INO1001, interferon, interleukin-12, ipilimumab, irinotecan, JNJ-16241199, ketoconazole, KRX-0402, lapatinib, lasofoxifene, LEE011, lectrozole, leucovorin, leuprolide, leuprolide , левамизол, заключенный в липосомы паклитаксел, ломустин, лонафарниб, люкантон, LY292223, LY292696, LY293646, LY293684, LY294002, LY3009120, LY317615, маримастат, мехлорэтамин, медроксипрогестеронацетат, магестролацетат, MEK162, мелфалан, меркаптопурин, месну, метотрексат, митрамицин, митомицин, mitotane, mitoxantrone, tosasertib, MLN8054, natitoclax, neovastat, neratinib, neuradiab, nilotinib, niluthimide, nolatrexed, NVP-BEZ235, oblimersen, octreotide, ofatumumab, oregovomab, ornatuzumab, orteronel, oxaliplatin, paclitaxel, palbociclib, pazopanumibum, pamidronate, PD0325901, PD184352, PEG interferon, pemetrexed, pentostatin, perifosine, phenylalanine mustard, PI-103, pictilisib, PIK-75, pipendoxifene, PKI-166, plicamycin, PLX8394, porfimer, predn ison, procarbazine, progestins, PX-866, R-763, raloxifene, raltitrexed, razoxin, ridaphorolimus, rituximab, romidepsin, RTA744, rubitecan, scriptaid, Sdx102, seliciclib, selume

- 6 039293 tinib, semaxinib, SF1126, sirolimus, SN36093, sorafenib, spironolactone, squalamine, SR13668, streptozocin, SU6668, suberoylanilide hydroxamic acid, sunitinib, synthetic estrogen, talampanel, talimogen lagerparevec, tamoxifen, temozolomide, shadow, tetrandrin, TGX-221, thalidomide, 6-thioguanine, thiotepa, ticilimumab, tipifarnib, tivozanib, TKI-258, TLK286, topotecan, toremifene citrate, trabectedin, tremetinib, trastuzumab, tretinoin, trichostatin A, triciribine phosphate monohydrate, triptorelin pamoate, TSE- 424, uracilmustard, valproic acid, valrubicin, vandetanib, vatalanib, VEGF decoy, vemurafenib, vinblastine, vincristine, vindesin, vinorelbine, vitaxin, vitespan, vorinostat, VX-745, wortmannin, Xr311, zanolimumab, ZK186619, ZK-303309, Z2 , ZSTK474, casopitant, netupitant, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylp rednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, GCSF, PEG-GCSF, erythropoietin, epoetin alfa and darbepoetin alfa. In one embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, is linked to pembrolizumab. In one embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, is linked to novolumab. In one embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, is linked to CT-011. In one embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, is linked to BMS936559.

The present invention also relates to a polypeptide containing an amino acid sequence selected from the group consisting of

SEQ ID NO: 2, 3, 4, 5, 7, 8, 9, 10, 12,13,

14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 27, 28, 29, 30, 32.33,

34, 35, 37, 38, 39, 40, 45, 47, 49, 51, 53, 55, 57, 59, 61.63,

65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93.95,

97, 99, 101, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146,148,

150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172.174,

176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198.200,

202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224,226,

228, 230, 232, 234, 235, 237, 239, 241, 243, 245, 247, 249.251,

253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275,277,

279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301,303,

305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327,329,

331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353,355,

357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379,381,

383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405,406,

407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418,419,

426, 427, 434, 435, 436, 437, 438, 439, 440, 441, 442, 446,448,

449, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461,462,

463 and 464, for example, SEQ ID NOs: 2, 7, 12, 17, 22, 27, 32 and 37;

or to its mature fragment. The present invention also relates to a polynucleotide encoding such polypeptides, for example, containing a nucleotide sequence selected from the group consisting of

SEQ ID NO: 1, 6, 11, 16, 21, 26, 31.36,

46, 48, 50, 52, 54, 56, 56, 58, 60, 62, 64, 66, 68, 70, 72.74,

76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127.129,

131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153.155,

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157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179.181,

183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205,207,

209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231,233,

236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258,260,

262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284,286,

288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310,312,

314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336,338,

340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362,364,

366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388,390,

392, 394, 396, 398, 400, 402 and 404;

or to its mature fragment.

Also included in the present invention is a vector (eg, plasmid) containing a polynucleotide. Also part of the present invention is a host cell (e.g., a mammalian cell, bacteria, Chinese hamster ovary (CHO), lower eukaryotes, fungi, yeast, Pichia, Pichia pastoris), where the host cell contains an antibody, a fragment, a polypeptide, a polynucleotide and/ or a vector described in the present description.

The present invention also relates to vaccines containing an antibody or fragment as described herein, an antigen (eg, viral peptide antigen, virus-like particle, tumor peptide antigen) and a pharmaceutically acceptable carrier.

Also, the present invention relates to a container (for example, a plastic or glass vial) or injection device (for example, a syringe, such as a pre-filled syringe or pen) containing any antibody, fragment, polypeptide, polynucleotide, vector, composition or vaccine described in this description.

The present invention also relates to a method of treating a malignant tumor (e.g., osteosarcoma, rhabdomyosarcoma, neuroblastoma, kidney cancer, leukemia, kidney transition cell cancer, bladder cancer, Wilms tumor, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer). , bone cancer, lung cancer, gastric cancer, colon and rectal cancer, cervical cancer, synovial sarcoma, head and neck cancer, squamous cell carcinoma, multiple myeloma, renal cell carcinoma, retinoblastoma, hepatoblastoma, hepatocellular carcinoma, melanoma, rhabdoid tumor of the kidney, Ewing's sarcoma, chondrosarcoma, malignant brain tumor, glioblastoma, meningioma, pituitary adenoma, vestibular schwannoma, primitive neuroectodermal tumor, medulloblastoma, astrocytoma, anaplastic astrocytoma, oligodenroglioma, ependymoma, papilloma of the choroid plexus, thrombocytomiopiaemia vera , sarcoma tissue, thyroid cancer, endometrial cancer, carcinoid or liver cancer, breast cancer, or gastric cancer) in an individual, comprising administering to the individual an effective amount of an anti-LAG3 antibody or antigen-binding fragment or vaccine described herein, optionally in conjunction with an additional drug or therapeutic procedure (for example, surgical tumorectomy or radiation therapy for cancer).

The present invention also relates to a method of administering an anti-LAG3 antibody, fragment, composition, polypeptide, vaccine, or polynucleotide described in the present invention, or a pharmaceutical composition thereof, to an individual, comprising injecting the antibody, fragment, polypeptide, vaccine, or polynucleotide into the body of the individual using an injecting device ; and optionally also administering an additional drug to the individual.

The present invention also relates to a method for producing an anti-LAG3 antibody, or an antigen-binding fragment thereof, or a polypeptide described in the present invention, comprising a) culturing a host cell containing a polynucleotide encoding a polypeptide or immunoglobulin chain of the antibody or fragment, in a culture medium under conditions contributing to the expression of the polynucleotide; and b) optionally, isolating the antibody, fragment, or polypeptide from the host cell and/or culture medium. In one embodiment of the invention, the method includes the step of introducing the polynucleotide into the host cell, for example, by transformation or transfection.

The present invention also relates to a method for detecting the presence of a LAG3 peptide or fragment thereof in a sample, comprising contacting the sample with an anti-LAG3 antibody or fragment described in the present invention and detecting the presence of a complex between the antibody or fragment and the peptide; where the detection of the complex indicates the presence of the LAG3 peptide.

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Brief description of the drawings

Fig. 1. Binding of anti-human LAG3 clones to CHO-K1 cells expressing human and cynomolgus LAG3.

Fig. 2. Binding of LAG3 clones to primary cynomolgus and human T cells.

Fig. 3. Three-dimensional structure of human LAG3. The position of the epitopes of antibodies 22D2, 11C9 and 4A10 is indicated.

Fig. 4 (A-C). Heatmap indicating regions in human LAG3 that are strongly or weakly protected from deuteration upon antibody binding. (A) heat map of differences for human LAG3/22D2; (B) heat map of differences for human LAG3/11C9; (C) Heat map of differences for human LAG3/4A10.

Fig. 5 (A-B). Combined heat maps indicating (A) regions of LAG3 that are protected from deuteration by binding to 22D2, 11C9 and 4A10, and (B) a position in human LAG3 mediating binding to MHC2.

Fig. 6. Predominant N-linked glycans for monoclonal antibodies produced in Chinese hamster ovary cells (CHO N-linked glycans) and engineered yeast cells (engineered yeast N-linked glycans): squares: N-acetylglucosamine (GlcNac) ; circles: mannose (Man); diamonds: galactose (Gal); triangles: fucose (Fuc).

Fig. 7. Effect of anti-human LAG-3 antibody treatment +/- anti-PD-1 antibody treatment on IL-2 production in SEB stimulated human PBMCs. PBMCs were activated with SEB for 3 days and the concentration of IL-2 in the culture supernatants was determined by MSD. Donor 090: SEB 60 ng/ml, Donor 089: SEB 30 ng/ml. The anti-PD1 agent was used at a concentration of 10 μg/ml.

Fig. 8. Effect of hu22D2 +/- treatment with anti-PD-1 antibody on IFN-γ and TNFa production in MLR stimulated human PBMCs. PBMCs were activated with allogeneic monocyte-derived DC for 7 days and the concentration of IFN-γ and TNF-α in culture supernatants was determined by MSD. The anti-PD-1 antibody was used at a concentration of 3 μg/ml. The isotype antibody was used at a concentration of 200 nM.

Fig. 9. Antibody capture assay to determine the concentration of unbound and partially bound anti-LAG3 Ab6 antibody in cynomolgus monkeys over time at various doses (0.03; 0.3; 1; 10; 30 mg/kg).

Fig. 10. Universal analysis of anti-LAG3 Ab6 total antibody concentration in cynomolgus monkeys over time at various doses (0.03; 0.3; 1; 10; 30 mg/kg).

Fig. 11. Dose-normalized Ab6 concentration over time in cynomolgus monkeys at various doses (0.03; 0.3; 1; 10; 30 mg/kg).

Fig. 12. Assessment of target-related clearance (Vmax, Km) of various doses of anti-LAG3 antibody Ab6.

Detailed description

The present invention relates to antibodies and antigen-binding fragments thereof that have exceptionally high affinity for human LAG3 and cynomolgus monkey LAG3, for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9, as well as their use and methods of obtaining, as described in the present description. For example, the measured affinity (KD) for human LAG3 by KinExA assay was as high as 2 pM and the affinity for cynomolgus monkey LAG3 was in the low two-digit pM range. In particular, the low isoelectric point (eg, approximately 6.3) makes some of these antibodies unique. Moreover, although some antibodies bind LAG3 primarily outside the region of the accessory loop, they demonstrate the ability to block class II LAG3/MCH binding. Moreover, the anti-LAG3 antibodies of the present invention have a high degree of specificity for LAG3 binding relative to other related proteins. Such antibodies and fragments described herein may be useful, for example, in the treatment of various cancers and infectious diseases.

Abbreviations.

In the detailed description and examples of the invention, the following abbreviations are used:

ADCC - antibody-dependent cellular cytotoxicity;

CDC - complement dependent cytotoxicity;

CDR - complementarity determining region in the variable regions of immunoglobulins, defined using the Kabat numbering system;

CHO - Chinese hamster ovary;

EC ₅₀ - concentration resulting in 50% efficiency or binding;

ELISA - enzyme-linked immunosorbent assay;

- 9 039293

FR - antibody framework region: variable regions of immunoglobulins, excluding CDR regions;

HRP - horseradish peroxidase;

IC50 - concentration leading to 50% inhibition;

IgG - immunoglobulin G;

Kabat is an immunoglobulin alignment and numbering system developed by Elvin A. Kabat ((1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.);

mAb or Mab or Mab is a monoclonal antibody;

PCR - polymerase chain reaction;

V-region - a segment of IgG chains, the sequence of which is variable between different antibodies. It continues to residue 109 in the light chain and 113 in the heavy chain according to Kabat;

VH - variable region of the heavy chain of immunoglobulin;

VK - immunoglobulin kappa light chain variable region.

Definitions.

To make the invention more understandable, certain technical and scientific terms are specifically defined below. Unless otherwise specifically indicated in this document, all other technical and scientific terms used in the present description have the meaning usually implied by a person skilled in the field to which the present invention pertains.

As used within the scope of the invention, including the appended claims, the singular form includes the plural of the respective entities referred to, unless the context clearly indicates otherwise.

LAG3.

The term LAG3, in relation to the polypeptide to which the antibodies and antigen-binding fragments of the present invention bind, refers to human and cynomolgus macaque LAG3, e.g., Macaca fascicularis or Macaca mulatta, as well as fragments thereof, such as its mature fragment lacking the signal peptide.

In one embodiment of the invention, the amino acid sequence of human LAG3 (lymphocyte activation gene 3) contains the amino acid sequence:

MWEAQFLGL FLQPLWVAPV KPLQPGAEVP WWAQEGAPA QLPCSPTIPL QDLSLLRRAG VTWQHQPDSG PPAAAPGHPL APGPHPAAPS SWGPRPRRYT VLSVGPGGLR SGRLPLQPRV QLDERGRQRG DFSLWLRPAR RADAGEYRAA VHLRDRALSC RLRLRLGQAS MTASPPGSLR ASDWVILNCS FSRPDRPASV HWFRRNRGQGR VPVRESPHHH LAESFLFLPQ VSPMDSGPWG CILTYRDGFN VSIMYNLTVL GLEPPTPLTV YAGAGSRVGL PCRLPAGVGT RSFLTAKWTP PGGGPDLLVT GDNGDFTLRL EDVSQAQAGT YTCHIHLQEQ QLNATVTLAI ITVTPKSFGS PGSLGKLLCE VTPVSGQERF VWSSLDTPSQ RSFSGPWLEA QEAQLLSQPW QCQLYQGERL LGAAVYFTEL SSPGAQRSGR APGALPAGHL LLFLILGVLS LLLLVTGAFG

FHLWRRQWRP

RRFSALEQGI HPPQAQSKIE ELEQEPEPEP ERERERER EPEQL (SEQ ID NO: 443); also see Uniprot accession number P18627.

In one embodiment, the mouse LAG3 amino acid sequence contains the amino acid sequence:

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MREDLLLGFL LLGLLWEAPV VSSGPGKELP WWAQEGAPV HLPCSLKSPN

LDPNFLRRGG

VIWQHQPDSG QPTPIPALDL HQGMPSPRQP APGRYTVLSV APGGLRSGRQ

PLHPHVQLEE

RGLQRGDFSL WLRPALRTDA GEYHATVRLP NRALSCSLRL RVGQASMIAS

PSGVLKLSDW

VLLNCSFSRP DRPVSVHWFQ GQNRVPVYNS PRHFLAETFL LLPQVSPLDS

GTWGCVLTYR

DGFNVSITYN LKVLGLEPVA PLTVYAAEGS RVELPCHLPP GVGTPSLLIA

KWTPPGGGPE

LPVAGKSGNF TLHLEAVGLA QAGTYTCSIH LQGQQLNATV TLAVITVTPK

SFGLPGSRGK

LLCEVTPASG KERFVWRPLN NLSRSCPGPV LEIQEARLLA ERWQCQLYEG

QRLLGATVYA

AESSSGAHSA RRISGDLKGG HLVLVLILGA LSLFLLVAGA FGFHWWRKQL

LLRRFSALEH

GIQPFPAQRK IEELERELET EMGQEPEPEP EPQLEPEPRQ L (SEQ ID NO: 444); also see Uniprot accession number Q61790. In one embodiment, the cynomolgus monkey LAG3 amino acid sequence comprises the amino acid sequence:

MWEAQFLGLL FLQPLWVAPV KPPQPGAEIS WWAQEGAPA QLPCSPTIPL

QDLSLLRRAG

VTWQHQPDSG PPAXAPGHPP VPGHRPAAPY SWGPRPRRYT VLSVGPGGLR

SGRLPLQPRV

QLDERGRQRG DFSLWLRPAR RADAGEYRAT VHLRDRALSC RLRLRVGQAS

MTASPPGSLR

TSDWVILNCS FSRPDRPASV HWFRSRGQGR VPVQGSPHHH LAESFLFLPH

VGPMDSGLWG

CILTYRDGFN VSIMYNLTVL GLEPATPLTV YAGAGSRVEL PCRLPPAVGT

QSFLTAKWAP

PGGGPDLLVA GDNGDFTLRL EDVSQAQAGT YICHIRLQGQ QLNATVTLAI

ITVTPKSFGS

PGSLGKLLCE VTPASGQEHF VWSPLNTPSQ RSFSGPWLEA QEAQLLSQPW

QCQLHQGERL

LGAAVYFTEL SSPGAQRSGR APGALRAGHL PLFLILGVLF LLLLVTGAFG

FHLWRRQWRP

RRFSALEQGI HPPQAQSKIE ELEQEPELEP EPELERELGP EPEPGPEPEP EQL (SEQ ID NO: 445); also see NCBI reference XP_005570011.1.

Mature human, mouse, and cynomolgus monkey LAG3 sequence, i.e. the sequence after removal of the signal peptide, contains amino acids 1-28 of SEQ ID NO: 443, 444 or 445.

The sequences of LAG3 may differ, for example, due to the presence of, for example, conserved mutations or mutations in non-conserved regions, for example, where LAG3 has essentially the same biological function as LAG3 SEQ ID NO: 443 or 445. For example, the biological functions of LAG3 are binding molecules of the major histocompatibility complex (MHC) class II and in the formation of homodimers.

The specific sequence of LAG3, as a rule, is at least 90% identical to the amino acid sequence of LAG3 SEQ ID NO: 443 or 445, or other isoforms. In certain instances, LAG3 may be at least 95%, or even at least 96%, 97%, 98%, or 99% identical to LAG3 of SEQ ID NO: 443 or 445, or other isoforms or variants. In certain embodiments, the LAG3 sequence shows no more than 10 amino acid differences from LAG3 according to any of SEQ ID NO: 443 or 445, or other isoforms or variants. In certain embodiments, LAG3 may show no more than 5, or even no more than 4, 3, 2, or 1 amino acid differences from LAG3 of SEQ ID NO: 443 or 445, or other isoforms or variants.

Percent identity can be determined as described herein.

Anti-LAG3 antibodies and their antigen-binding fragments.

The present invention relates to antibodies or antigen-binding fragments thereof which specifically bind LAG3 (e.g. human and/or cynomolgus monkey LAG3, e.g. Macaca fascicularis or Macaca mulatta), e.g. 4A10, 19E8, 11C9, 22D2, e.g. Ab1, Ab2, Ab3, Ab4, Ab5,

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Ab6, Ab7, Ab8 or Ab9, and the use of such antibodies or fragments. In one embodiment, the antibody or fragment is an antibody.

As used herein, an anti-LAG3 antibody, or antigen-binding fragment thereof, refers to an antibody, or antigen-binding fragment thereof, that specifically binds to human or cynomolgus monkey LAG3. An antibody specifically binds to a polypeptide containing the sequence (in this case, a human or cynomolgus LAG3 epitope) if it binds to polypeptides containing the LAG3 sequence with a K _D of approximately 1 nM or higher affinity (e.g., 1 nM-2 pM, 1 nM, 100 pM, 10 or 2 pM), but does not bind to proteins lacking this sequence. For example, an antibody or antigen-binding fragment that specifically binds a polypeptide containing human or cynomolgus macaque LAG3 can bind to a FLAG®-tagged form of human or cynomolgus monkey LAG3, but does not bind to other FLAG®-tagged proteins that lack LAG3 epitopes.

The present invention relates to anti-LAG3 antibodies and methods for their use. As used herein, the term antibody refers to any form of an antibody that exhibits the desired biological activity. Thus, it is used in its broadest sense and specifically includes, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, polyspecific antibodies (e.g., bispecific antibodies), humanized antibodies, fully human antibodies, chimeric antibodies and camelized single domain antibodies.

The present invention relates to parental antibodies against LAG3 and their antigen-binding fragments, and to methods of their use. Parental antibodies and antigen-binding fragments thereof are antibodies and fragments that can be modified for their intended use, such as humanizing the antibody for use as a therapeutic antibody or fragment in humans.

The present invention relates to antigen-binding fragments of LAG3 and to methods of their use. As used herein, unless otherwise indicated, an antibody fragment or antigen-binding fragment refers to antigen-binding antibody fragments, i.e. fragments of an antibody that retain the ability to specifically bind to an antigen bound by a full-length antibody, eg, fragments that retain one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2 and Fv fragments; diantibodies; single chain antibody molecules, eg sc-Fv; nanoantibodies and polyspecific antibodies formed from antibody fragments.

The present invention relates to Fab fragments against LAG3 and to methods of their use. The Fab fragment consists of one light chain and CH1 and one heavy chain variable regions. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule. The Fab fragment may be a cleavage product of an antibody with papain.

The present invention relates to antibodies against LAG3 and their antigennegative fragments, which contain the Fc region, and to methods of their use. The Fc'' region contains two heavy chain fragments containing the CH1 and CH2 domains of the antibody. The two heavy chain fragments are held together by two or more disulfide bonds and hydrophobic interactions of the CH3 domains.

The present invention relates to Fab'-fragments against LAG3 and to methods of their use. A Fab' fragment contains one light chain and a portion or fragment of one heavy chain that contains a VH domain and a CH1 domain, as well as a region between the CH1 and CH2 domains, so that an interchain disulfide bond can form between the two heavy chains of the two Fab' fragments with the formation of the F(ab') ₂ molecule.

The present invention relates to F(ab') ₂ fragments against LAG3 and to methods of their use. The F(ab') ₂ fragment contains two light chains and two heavy chains containing part of the constant region between the CH1 and CH ₂ domains so that a disulfide bond is formed between the two heavy chains. Thus, the F(ab') ₂ fragment consists of two Fab' fragments that are held together by a disulfide bond between the two heavy chains. The F(ab')2 fragment may be the product of cleavage of the antibody with pepsin.

The present invention relates to anti-LAG3 Fv fragments and methods of using them. The Fv region contains both heavy and light chain variable regions, but lacks constant regions.

The present invention relates to anti-LAG3 scFv fragments and methods for their use. The term single chain Fv or scFv antibody refers to antibody fragments containing the VH and VL domains of an antibody, these domains being in the same polypeptide chain. Typically, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that allows the scFv to form the desired structure for antigen binding. For a review of scFv, see Pluckthun (1994) THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and Moore eds. SpringerVerlag, New York, p. 269-315. See also International Patent Application Publication No. WO 88/01649 and

- 12 039293 US patents No. 4946778 and 5260203.

The present invention relates to domain antibodies against LAG3 and to methods of their use. A domain antibody is an immunologically functional fragment of an immunoglobulin containing only a heavy chain variable region or a light chain variable region. In some cases, two or more VH regions are covalently linked by a peptide linker to form a bivalent domain antibody. The two VH regions of a bivalent domain antibody may be directed to the same or to different antigens.

The present invention relates to bivalent antibodies against LAG3 and to methods of their use. A bivalent antibody contains two antigen-binding centers. In some cases, two binding centers have the same antigenic specificity. However, bivalent antibodies can be bispecific (see below).

The present invention relates to camelized anti-LAG3 single domain antibodies and methods of using the same. In certain embodiments, the antibodies described herein also include camelized single domain antibodies. See, for example, Muyldermans et al. (2001) Trends Biochem. sci. 26:230; Reichmann et al. (1999) J. Immunol. Methods 231:25; W094/04678; W094/25591; US patent No. 6005079). In one embodiment, the present invention relates to single domain antibodies containing two VH domains with modifications to form single domain antibodies.

The present invention relates to anti-LAG3 diabodies and methods for their use. As used herein, the term diabody refers to small antibody fragments with two antigen-binding sites that contain a heavy chain variable domain (VH) linked to a light chain variable domain (VL) in the same polypeptide chain (VH-V _L or VLVH). Using a linker that is too short to allow pairing between two domains on the same strand, the domains are forced to pair with complementary domains on the other strand and form two antigen binding sites. Diabodies are described in more detail, for example, in EP 404097; WO 93/11161; and Holliger et al. (1993) Proc. Natl. Acad. sci. USA 90: 64446448. For a review of engineered antibody variants, especially see Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

Typically, an antibody or antigen-binding fragment of the invention that is modified in some way retains at least 10% of its LAG3 binding activity (e.g., human and/or cynomolgus monkey LAG3, e.g., Macaca fascicularis or Macaca mulatta) (compared to the parent antibody) when this activity is expressed on a molar basis. Preferably, the antibody or antigen-binding fragment of the invention retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the parent antibody's binding affinity for LAG3 (e.g., human and/or cynomolgus monkey LAG3, e.g., Macaca fascicularis or Masa mulatta). It is also contemplated that an antibody or antigen-binding fragment of the invention may include conservative or non-conservative amino acid substitutions (referred to as conservative variants or functionally conservative variants of an antibody) that do not substantially alter its biological activity.

The present invention provides isolated anti-LAG3 antibodies and antigen-binding fragments thereof (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, and/or Ab9) and methods their uses, as well as their isolated polypeptide immunoglobulin chains and isolated polynucleotides encoding such polypeptides, and isolated vectors comprising such polynucleotides. The isolated antibodies or their antigen-binding fragments, polypeptides, polynucleotides and vectors are at least partially free from other biological molecules of the cell or cell culture from which they are derived. Such biological molecules include nucleic acids, proteins, lipids, carbohydrates, or other material such as cell debris and growth media. The isolated antibody, or antigen-binding fragment thereof, may also be at least partially free of components of the expression system, such as biological molecules from the host cell or their growth media. Generally, the term isolated does not refer to the complete absence of such biological molecules, or to the absence of water, buffers or salts, or components of a pharmaceutical formulation that includes antibodies or fragments.

The present invention relates to anti-LAG3 monoclonal antibodies and antigen-binding fragments thereof, as well as to monoclonal compositions containing a plurality of isolated monoclonal antibodies. The term monoclonal antibody, as used herein, refers to a population of substantially homogeneous antibodies, i.e. the antibody molecules that make up the population are identical in amino acid sequence, with the exception of possible naturally occurring mutations, which may be present in small quantities. The plurality of such monoclonal antibodies and fragments in a composition refers to the concentration of identical (i.e., as discussed above, in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts) antibodies and

- 13 039293 fragments, which exceeds the concentration normally found in nature, for example, in the blood of a host organism such as a mouse or a human. In contrast, conventional preparations of (polyclonal) antibodies typically include many different antibodies having different amino acid sequences in their variable domains, in particular their CDRs, which are often specific for different epitopes. The definition of monoclonal indicates that the characteristic of an antibody is that it is derived from a substantially homogeneous population of antibodies and should not be construed as requiring the production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention can be obtained by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or they can be obtained by recombinant DNA methods (see, for example, US patent No. 4816567). Monoclonal antibodies can also be isolated from phage antibody libraries using the methods described, for example, in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222:581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

The present invention relates to chimeric anti-LAG3 antibodies (eg, human constant domain/mouse variable domain) and methods of using the same. As used herein, a chimeric antibody is an antibody having a variable domain from a first antibody and a constant domain from a second antibody, wherein the first and second antibodies are from different species (US Pat. No. 4,816,567; and Morrison et al. (1984) Proc. Natl Acad Sci USA 81: 6851-6855). Typically, variable domains are derived from an experimental animal antibody (parent antibody), such as a rodent, and constant domain sequences are derived from human antibodies, such that the resulting chimeric antibody is less likely to induce an adverse immune response in humans than the parent antibody (e.g., mice).

The present invention relates to humanized anti-LAG3 antibodies and antigen-binding fragments thereof (eg, mouse antibodies that have been humanized) and methods of using them. As used herein, the term humanized antibody refers to forms of antibodies that contain sequences from both human and non-human (eg, mouse or rat) antibodies. Typically, a humanized antibody contains substantially all of at least one, and typically two, variable domains, wherein all or substantially all of the hypervariable loops correspond to non-human immunoglobulin hypervariable loops, and all or substantially all of the framework (FR) regions are the framework regions of a human immunoglobulin sequence. The humanized antibody may optionally comprise at least a portion of a human immunoglobulin constant region (Fc).

The present invention relates to fully human anti-LAG3 antibodies and antigen-binding fragments thereof, and methods of using them. The term fully human antibody refers to an antibody that contains only human immunoglobulin protein sequences. A fully human antibody may contain mouse carbohydrate chains if it is produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a mouse antibody refers to an antibody that contains only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain rat carbohydrate chains if produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, a rat antibody refers to an antibody that contains only rat immunoglobulin sequences. In one embodiment, the fully human anti-LAG3 antibody or antigen-binding fragment thereof is a product isolated from a transgenic animal, e.g., a mouse (e.g., a HUMAB mouse, see e.g., US Pat. Nos. 5,545,806; 5,569,825; 5,625,126; ; 5789650; 5814318; 5874299 and 5877397; and Harding, et al. (1995) Ann. NY Acad. Sci. 764:536 546; or XENOMOUSE, see, for example, Green et al., 1999, J. Immunol. Methods 231:11-23) that is genetically modified to have fully human immunoglobulin genes; or a product isolated from a phage or virus that expresses the immunoglobulin chains of a fully human anti-LAG3 antibody or antigen-binding fragment thereof.

As a rule, the main structural element of the antibody contains a tetramer. Each tetramer includes two identical pairs of polypeptide chains, with each pair having one light (approximately 25 kDa) and one heavy chain (approximately 50-70 kDa). The N-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids, primarily responsible for antigen recognition. The C-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Generally, human light chains are classified as kappa and lambda light chains. Moreover, human heavy chains are generally classified as mu, delta, gamma, alpha, or epsilon, and they define the antibody isotype as IgM, IgD, IgG, IgA, and IgE, respectively. In the light and heavy chains, the variable and constant regions are linked by a J region of about 12 or more amino acids, with the heavy chain also including a D region of about 10 or more amino acids. See mainly Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989).

- 14 039293

In one embodiment, the anti-LAG3 antibodies of the present invention comprise a complete tetrameric structure having two light chains and two heavy chains, including constant regions.

The variable regions of each light/heavy chain pair form the antibody binding site. Thus, typically, an intact antibody has two binding sites. With the exception of bifunctional or bispecific antibodies, the two binding sites are usually the same.

Typically, both heavy and light chain variable domains contain three hypervariable regions, also referred to as complementarity determining regions (CDRs), located in relatively conserved framework regions (FRs). The CDRs are usually arranged parallel to the framework regions to allow binding to a specific epitope. Typically, from the N-terminus to the C-terminus, the variable domains of both the light and heavy chains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each domain is generally carried out in accordance with the definitions of Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5th ed ^.; NIH Publ. no. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat, et al, (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al, (1987) J Mol. Biol. 196:901-917 or Chothia, et al. (1989) Nature 342:878-883.

As used herein, the term hypervariable region refers to the amino acid residues of an antibody, or antigen-binding fragment thereof, that are responsible for antigen binding. The hypervariable region contains amino acid residues from the complementarity determining region or CDR (ie CDRL1, CDRL2 and CDRL3 in the light chain variable domain and CDRH1, CDRH2 and CDRH3 in the heavy chain variable domain). See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th ^Ed . Public Health Service, National Institutes of Health, Bethesda, Md.; Johnson et al. (2001) Nucleic Acids Res. 2001; 29(1): 205-206 (which defines antibody CDR regions by sequence); also see Chothia and Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al., Nature 342, 877 (1989), and Tramontano et al., J. Mol. Biol. 215, 175 (1990) (which defines antibody CDR regions by structure); also see Macallum et al., J Mol Biol. 1996 Oct. eleven; 262(5):732-45. As used herein, the term framework residues or FR residues refers to variable domain residues other than hypervariable region residues, defined herein as CDR residues.

The scope of the present invention includes anti-LAG3 antibodies and antigen-binding fragments thereof that specifically bind LAG3, which may have any combination of CDRs from immunoglobulin light chains of SEQ ID NOs: 7, 17, 27 and/or 37, and/or which have any combination of CDRs from immunoglobulin heavy chains SEQ ID NOs: 2, 12, 22 and 32, wherein the CDRs are as defined by Kabat and Chothia (see above).

Homology refers to the sequence similarity between two polynucleotide sequences or between two polypeptide sequences when they are optimally aligned. If a position in both of the two compared sequences is occupied by the same base or monomeric amino acid subunit, for example if a position in each of the two DNA molecules is occupied by adenine, then the molecules are homologous at that position. Percent homology is the number of positions that are common to two sequences divided by the total number of compared positions x100. For example, if 6 out of 10 positions in two sequences match or are homologous, when the sequences are optimally aligned, then the two sequences are 60% homologous. Typically, the comparison is made when the two sequences are aligned to obtain the maximum percent homology.

The isolated nucleic acid molecules or isolated polynucleotides (eg, DNA or RNA) are also not associated with all or part of the polynucleotide in which the isolated polynucleotide occurs naturally, or associated with a polynucleotide with which they are not naturally associated. For the purposes of the present invention, it should be understood that a nucleic acid molecule containing a particular nucleotide sequence does not span intact chromosomes.

Isolated nucleic acid molecules containing said nucleic acid sequences may include, in addition to said sequences, coding sequences for up to ten or even up to twenty or more other proteins or regions or fragments thereof, or they may include operably linked regulatory sequences, which control the expression of the coding region of said nucleic acid sequences, and/or they may include vector sequences. As discussed below, the present invention includes isolated polynucleotides encoding any of the immunoglobulin chains described in the present invention.

Expression control sequences refer to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. Control sequences that are useful in prokaryotes include, for example, a promoter, optionally an operator sequence, and a ribosome binding site. It is known

- 15 039293 that eukaryotic cells use promoters, polyadenylation signals and enhancers.

A nucleic acid or polynucleotide is operably linked when it is in a functional relationship with another nucleic acid sequence. For example, the DNA for a presequence or secretory leader is operably linked to the DNA of a polypeptide if it is expressed as a preprotein that is involved in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned to promote translation. In general, operably linked means that the linked DNA sequences are adjacent, and in the case of a secretory leader sequence, adjacent and in reading frame. However, enhancers do not need to be adjacent. Binding is carried out by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers are used in accordance with common practice.

As used herein, the terms cell, cell line, and cell culture are used interchangeably, and all such designations include progeny. Thus, the words transformants and transformed cells include the primary cells of the individual and the cultures derived from them, regardless of the number of transfers. It should also be understood that not all offspring have exactly identical DNA content due to intentional or accidental mutations. It includes mutant progeny that have the same function or biological activity as screened in the originally transformed cell. When certain designations are intended, this will be apparent from the context.

As used herein, polymerase chain reaction or PCR refers to a technique or process in which certain nucleic acid sequences, RNA and/or DNA, are amplified as described, for example, in US Pat. No. 4,683,195. Generally, to design oligonucleotide primers use sequence information from the ends of the region of interest or beyond. These primers are identical or similar in sequence to the opposite strands of the template to be amplified. The 5'-terminal nucleotides of the two primers may coincide with the ends of the amplified material. PCR can be used to amplify specific RNA sequences, specific DNA sequences from total genomic DNA and cDNA transcribed from total cellular RNA, bacterophage or plasmid sequences, and so on. See especially Mullis et al. (1987) Cold Spring Harbor Symp. quant. Biol. 51:263; Erlich, ed. (1989) PCR TECHNOLOGY (Stockton Press, N.Y.). As used herein, PCR is considered one, but not the only, example of a nucleic acid polymerase reaction method for amplifying a nucleic acid sample of interest, comprising using a known nucleic acid as a primer and a polymerase to amplify the nucleic acid or obtain a specific nucleic acid fragment.

As used herein, a germline sequence refers to unrearranged immunoglobulin DNA sequences. Any suitable source of unrearranged immunoglobulin sequences may be used. Human germline sequences can be obtained, for example, from the JOINSOLVER germline sequence databases at the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the United States National Institutes of Health website. Mouse germline sequences can be obtained, for example, as described in Giudicelli et al. (2005) Nucleic Acids Res. 33:D256-D261.

Physical and Functional Properties of Exemplary Anti-LAG3 Antibodies

The present invention provides anti-LAG3 antibodies and antigen-binding fragments thereof (eg, humanized antibodies, such as humanized antagonist antibodies) and methods of using antibodies or antigen-binding fragments thereof to treat or prevent a disease. In one embodiment, the invention relates to anti-mouse LAG3 antibodies or humanized anti-LAG3 antibodies and antigen-binding fragments thereof, and methods of using the antibodies or antigen-binding fragments thereof to treat or prevent a disease. In one embodiment, the invention relates to anti-LAG3 antagonist antibodies and methods of using the antibodies, or antigen-binding fragments thereof, to treat or prevent a disease.

Within the scope of the present invention, an anti-LAG3 antibody or antigen-binding fragment thereof comprising a particular light chain and a particular heavy chain may be referred to as light chain/heavy chain; for example, an antibody containing the light chain 45AGX_22D2_VL3 and the heavy chain Humashzedχ[LAG3_H] mAb.22D2 VH6 N54D can be designated as

45AGX_22D2_VL3/Humanizedx[LAG3_H] mAb.22D2 VH6 N54D.

An anti-LAG3 antibody or antigen-binding fragment thereof of the present invention includes any antibody or antigen-binding fragment thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8

- 16 039293 and/or Ab9) that specifically bind to LAG3 (eg human or cynomolgus monkey LAG3). Such antibodies and fragments include humanized antibodies and fragments having any combination of mouse light and heavy chains and humanized light and heavy chains as described herein, or variants of such chains that specifically bind LAG3. Such antibodies and fragments include any antibody or fragment containing any one or more of the CDRs (e.g., CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) of murine or humanized chains, described in the present invention, or variants of such CDRs that specifically bind LAG3. Moreover, an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention includes any antibody or antigen-binding fragment thereof that binds to the same epitope in LAG3 to which the antibodies and fragments described herein bind, and any antibody or antigen-binding fragment that cross-block (partially or completely) or cross-blocked (partially or completely) with an antibody or fragment described in the present invention, in relation to the binding of LAG3; as well as to any of their options. A specific embodiment of the invention includes antibodies and fragments containing only mouse immunoglobulin chains or only humanized immunoglobulin chains and/or where all of the immunoglobulin chains or CDRs are derived, directly or indirectly, from the same parent mouse clone, i. humanized 4A10 light chains paired with humanized 4A10 heavy chains; humanized 19E8 light chains paired with humanized 19E8 heavy chains; humanized 11C9 light chains paired with humanized 11C9 heavy chains; or humanized 22D2 light chains paired with humanized 22D2 heavy chains; or 4A10 mouse light chains paired with 4A10 mouse heavy chains; 19E8 mouse light chains paired with 19E8 mouse heavy chains; 11C9 mouse light chains paired with 11C9 mouse heavy chains; or mouse 22D2 light chains paired with 22D2 mouse heavy chains. These antibodies and fragments are part of the present invention, as well as their use, for example, as described in the present description.

Cross-blocking antibodies and their antigen-binding fragments described herein can be identified based on their ability to block any of the antibodies or fragments specifically mentioned herein, e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7 , Ab8, Ab9, 4A10, 19E8, 11C9, and/or 22D2, for binding to LAG3, in binding assays (e.g., biolayer interferometry (BLI; e.g., ForteBio Octet binding assay; Pall ForteBio Corp; Menlo Park, CA), analysis using surface plasmon resonance (SPR), BIACore, ELISA, flow cytometry). For example, in one embodiment of the invention using BLI, the fiber optic probe tip is coated with a ligand (e.g., LAG3) and acts as a biosensor where binding of an anti-LAG3 antibody or antigen-binding fragment thereof to LAG3 changes the white light interference pattern reflected from the layer probe associated with LAG3 and an internal reference layer. The shift indicates binding of LAG3/Anti-LAG3 antibody. In one embodiment, the LAG3-coated tip is immersed in a solution of the test material containing the antibody or antigen-binding fragment, for example, in a well of either a 96-well or 384-well plate. In one embodiment of the invention, the tablet is shaken during reading to create an orbital flow. For reading in the assay, white light is directed down the length of the fiber. As mentioned above, interference between light reflected from the reference layer and immobilized surfaces containing LAG3 at the tip creates a distinct pattern of light returning upward along the fiber. When molecules bind to an immobilized sensory surface, this pattern changes in proportion to the degree of binding. For example, assays can be used in which a LAG3 protein (e.g., human LAG3) is immobilized on a BLI probe or plate, a reference anti-LAG3 antibody or fragment is coupled to LAG3 (e.g., at saturating concentration), and the anti-LAG3 antibody or fragment of interest is added. The ability of the test antibody to compete with the reference antibody for LAG3 binding is then determined. The BLI format monitors light interference for the LAG3 complex to determine if the antibody of interest competes effectively with the reference antibody, e.g. nanometer shift in light wavelength over time, where the shift indicates additional binding of the antibody of interest and no cross-blocking. In one embodiment of the invention in the BLI format, the presence of cross-blocking between antibodies is qualitatively determined if no additional binding of the test antibody is observed. In one embodiment of the invention, as a control, cross-blocking of the reference antibody against itself is confirmed; where the assay is considered to be functioning properly if the reference antibody can cross-block its own binding to LAG3. The ability of the test antibody to inhibit the binding of, for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, 4A10, 19E8, 11C9, and/or 22D2 to LAG3 (e.g., human LAG3) demonstrates that the test antibody can cross-block Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, 4A10, 19E8, 11C9, and/or 22D2 for binding to LAG3 (e.g., human LAG3) and thus, in some cases, may bind to the same epitopes on LAG3 (eg, human LAG3) as Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, 4A10, 19E8, 11C9 and/or

- 17 039293

22D2. As indicated above, antibodies and fragments that bind to the same epitope as any of the anti-LAG3 antibodies or fragments of the present invention are also part of the present invention. In one embodiment, BLI is carried out in a sandwich format, where a reference anti-LAG3 antibody or antigen-binding fragment is immobilized on a probe and then bound to LAG3. The test LAG3 antibody or antigen-binding fragment thereof is then tested for its ability to block the binding of the reference antibody or fragment.

Anti-LAG3 antibodies 4A10, 19E8, 11C9, and 22D2 and antigen-binding fragments thereof, as used herein, include CDR-L1, CDR-L2, and CDR-L3 of mouse immunoglobulin light chains 4A10, 19E8, 11C9, or 22D2, and variants thereof, respectively; as well as CDR-H1, CDR-H2 and CDR-H3 of heavy chains of mouse immunoglobulins 4A10, 19E8, 11C9 or 22D2, and their variants, respectively. Such 4A10, 19E8, 11C9 and 22D2 antibodies and fragments thereof may be humanized antibodies or antigen-binding fragments thereof such as Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9.

Examples of immunoglobulin chains in anti-LAG3 antibodies, as well as their CDRs, include, but are not limited to:

4A10 - VH sequence

ATGAAATGCAGCTGGGTCATCTTCTTCCTGATGGCAGTGGTTATAGGAATCAATTCAGA

GGTTCAGCTGCTCCAGTCTGGGGCAGAACTTGTGAGGTCAGGGGCCTCAGTCAAGTTGTCCTGC

ACAGC C T C T GGC T T CAACAT T GAAGAC TAC TATAT GCAC TGGAT GAAACAGAGGCC T GAACAGG

GCCTGGAGTGGATTG GATGGATTGATCCTGTGAATGGTGATACTGAATATGCCCCGAAGTTCCA GGGCAAGGCCACTATGACTGCAGACACATCCTCCAACACAGCCTACCTACACCTCAACAGCCTG АСАТ С Т GAGGACAC Т GCCGT С ТАТ ТАС Т G ТААТ Т Т CTATGATGGTTACCTCTTTGCTTTCTGGG GCCAAGGGACCCTGGTCACTGTCTCTGCA (SEQ ID NO: 1; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом)

MKCSWVIFFLMAWIGINSE VQLLQS GAE LVRS GAS VKLS WITH TAS GFNIEDYYMHWMKOR

PEOGLEWIGWIDPVNGDTEYAPKFQGKATMTADTSSNTAYLHLNSLTSEDTAVYYCNFYDGYLF

AFWGOGTLVTVSA (SEQ ID NO: 2; where CDRs are underlined and where the signal sequence is in bold)

CDR-H1: GFNIEDYYMH (SEQ ID NO: 3)

CDR-H2: WIDPVNGDTEYAPKFQG (SEQ ID NO: 4)

CDR-H3: YDGYLFAF (SEQ ID NO: 5)

4A10 - sequence V _L

ATGAGGTGCCTAGCTGAGTTCCTGGGGCTGCTTGTGCTCTGGATCCCTGGAGCCATTGG

GGATATTGTGCTGACTCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTGTCCATC

T CC T GCAGGTCTAGTAAGAGTCTCCTGCATAGTGATGGCAACACTTATCTGTATT GGCTCCTGC AGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGGGTCCC AGACAGGTTCAGCGGCAGTGGGTCAGGAACTGTTTTCACACTGAGAATCAGCAGACTGGAGGCT GAGGAT GT GGG TAT T TAT ТАС T G T AT GCAACAT С T AGAAT AT С С T T T CAC G T T T GGAGGGGGGA CCAAGCTGGAAATAAAA (SEQ ID NO: 6; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом)

MRCLAE FLGLLVLWIPGAIGDIVL TQAAP SVPVT PGE SVSIS CRSSKSLLHSDGNTYLY

WL LORPGOSPOLLIYRMSNLASGVPDRFSGSGSGTVFTLRISRLEAEDVGIYYCMQHLEYPFTF GGGTKLEIK (SEQ ID NO: 7; where CDRs are underlined and where the signal sequence is in bold)

CDR-L1: RSSKSLLHSDGNTYLY (SEQ ID NO: 8)

CDR-L2: YRMSNLAS (SEQ ID NO: 9)

CDR-L3: MQHLEYPFT (SEQ ID NO: 10)

19E8 - VH sequence

- 18 039293

ATGGGATGGAGCTGGATCTTTCTTTTCCTCCTGTCAGGAACTGCAGGTGTCCGTTGCCA

GATCCGACTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGC AAGGC T T С TGGGTCCTCCTTCACTGACTACTATATAAACT GGGT GAAGCAGAAGCC T GGACAGG GAC T T GAGT GGAT TGGATGGATTTATCCTGGAAGCGGTAATTCTATCTACAATGAGAACTTCAA GGCCAAGGCCACATTGACTGTAGACACATCCTCCAGCACAGCCTACATGCATCTCAGCAGCCTG ACAT С T GAGGACAC T GC T GT C TAT T T C TGTGCAAGAGAGGCTGATTACGACGATGCTTTGGACT ACTGGGGTCAAGGAACCTCGGTCACCGTCTCCTCA (SEQ ID NO: 11; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом)

MGWSWIFLFLLSGTAGVRCQIRLQQS G PE LVKPGASVKIS CKASGSSFTDYYINWVKQK PGQGLEWIGWIYPGSGNSIYNENFKAKATLTVDTSSS TAYMHLSSLTSEDTAVYFCAREADYDD ALDYWGOGTSVTVSS (SEQ ID NO: 12; where the CDRs are underlined and where the signal sequence is in bold)

CDR-H1: GSSFTDYYIN (SEQ ID NO: 13)

CDR-H2: WIYPGSGNSIYNENFKA (SEQ ID NO: 14)

CDR-H3: EADYDDALDY (SEQ ID NO: 15)

19E8 - sequence V _L

ATGGTATCCACACCTCAGTTCCTTGTATTTTTGCTTTTCTGGATTCCAGCCTCCAGAGG

TCACATCTTGCTGACTCAGTCTCCAGCCATTCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTC T С С T G CAGGGCCAGTCAGAGCATTGGCACAAGCATACACTGGTAT C AGCAAAGAACAAAT GG T T CTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATCCCTTCCAGGTTTAGTGG CAG T GGAT CAG G GACAGAT T T TAC T С T TAG CAT СAACAG T G T GGAG T CAGAAGATAT T GCAGAT TATTACTGTCAACAAAGTAATAGCTGGCCAACGTACACGTTCGGAGGGGGGACCAAGCTGGAAA TAAAA (SEQ ID NO: 16; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом )

MVSTPQFLVFLLFWIPASRGHILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQR

TNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQOSNSWPTYTFGGGT KLEIK (SEQ ID NO: 17; where CDRs are underlined and where the signal sequence is in bold)

CDR-L1: RASQSIGTSIH (SEQ ID NO: 18)

CDR-L2: YASESIS (SEQ ID NO: 19)

CDR-L3: QQSNSWPTYT (SEQ ID NO: 20)

11C9 - sequence V _H

ATGAGATGGAGC TGTATCATCC TCTTC T TGGTAGCAACAGCTACAGGTGTCAAC TCCC A

GGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGATGCCTGGGGCTTCAGCGAAGATGTCCTGC AAGGCTTCTGGCTACACACTCACTGACTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAG GCCTTGAGTGGATCG GAG C GAT T GATAT T T С T GATAG T TAT T С TAGC TACAAT CAAAAG T T СAA GGGCAAGGCCACATT GAC T G TAGAC GAAT CC T C CAGCACAGC С TACAT GCAGC T CAC CAGCC T G ACAT С T GAGGAC TCTGCGGTC TAT ТАС T GT GCAAGATCCCCTTTCTACAATAGTAGAGGGGGGA AC TAC T T T GAC TAC TGGGGC CAAGGCAC CAC T C T CACAGT C T C C T CA (SEQ ID NO: 21; where CDRs are underlined and where the signal sequence is in bold)

MRWSC11LFLVATATGVNSQVQL QQ P GAE LVHP GASAKMS CKASGYTLTDYWMHWVKQR PGQGLEWIGAIDISDSYSSYNQKFKGKATLTVDES S S TAYMOLTSLTSEDSAVYYCARSPFYNS RGGNYFDYWGOGTTLTVSS (SEQ ID NO: 22; where the CDRs are underlined and where the signal sequence is in bold)

CDR-Hl: GYTLTDYWMH (SEQ ID NO: 23)

CDR-H2: AIDISDSYSSYNQKFKG (SEQ ID NO: 24)

CDR-H3: SPFYNSRGGNYFDY (SEQ ID NO: 25)

- 19 039293

11C9 - VL sequence

ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGGTACCAGATG

T GATAT С CAGAT GAGACAGAC TAGAT CCTCCCTGTCTGCCTCTCTGG GAGACAGAG T GAG CATC AGTTGCAGGGCAAGTCAGGACATTAGCAATTATTTAAACTGGTAT CAGCAGAAACCAGAT GGAA С T G T TAAAC T С С T GAT С ТАС TACACAT CAAGAT TACAC T CAGGAG T С С CAT CAAGG T T CAG T GG CAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACT ТАС T T T TGCCAACAGGGTGATACGCTTCCTCCGTGGACGTT CGGT GGAGGCACCAAGC T GGAAA TCAAA (SEQ ID NO: 26; where CDRs are underlined and where the signal sequence is in bold)

MMSSAQFLGLLLLCFQGTRCDIQMT QT S S L SAS LGDRVTIS CRASQDISNYLNWYQQK PDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSIjTISNLEOEDIATYFCQQGDTLPPWTFGGGT KLEI K (SEQ ID NO: 27; where CDRs are underlined and where the signal sequence is in bold)

CDR-L1: RASQDISNYLN (SEQ ID NO: 28)

CDR-L2: YTSRLHS (SEQ ID NO: 29)

CDR-L3: QQGDTLPPWT (SEQ ID NO: 30)

22D2 - sequence V _H

ATGGGATGGACCTGGATCTTTCTCTTCTTCCTGTCAGGAACTGCAGGTGTCCTCTCTGA

GGTCCTGCTGCTACAGTCTGGACCTGAACTGGTGAAGCCTGGGACTTCAGTGAAAATCCCCTGC AAGGC T T C TGGATACACATTCACTGACTACAACGTGGACTGGGTGAAGCAGCGCCATGGAAAGG GC С T T GAGT GGAT T G GAGATAT TAAT С CAAACAAT GGTGGTACTATC TAGAG T CAGAAAT T СAA GGGCAAGGCCACATTGACTGTTGACAAGTCCTCCAGCACAGCCTTCATGGAGCTCCGCAGCCTG ACAT С T GAGGACAC T GCAG T C TAT T T C TGTGCAAGGAACTATAGGTGGTTTGGTGCTATGGACC ACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAACAGCCCCATCGGTCTATCC ACTG (SEQ ID NO: 31; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом )

MGWTWIFLFFLSGTAGVLSEVLLLQSGPELVKPGTSVKIPCKASGYTFTDYNVDWVKQR

HGKGLEWIGDINPNNGGTIYSQKFKGKATL TVDKS S S TAFME LRSL T S E DTAVY FCARNYRWFG

AMDHWGOGTSVTVSS (SEQ ID NO: 32; where CDRs are underlined and where the signal sequence is in bold)

CDR-Hl: DYNVD (SEQ ID NO: 33)

CDR-H2: DINPNNGGTIYSQKFKG (SEQ ID NO: 34)

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35)

22D2 - VL sequence

ATGGAGACAGACACAATCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGGTTCCACTGG

TGACATTGTGTTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCCAGGGCAGAGGGCCACCATT T С С T G CAAG G C CAG T CAAAG TCTTGATTAT GAAG G T GATAG T GATAT GAAT TGGTACСAACAGA AACCAGGACAGCCACCCAGACTCCTCATCTCTGGTGCATCCAATCTAGAGTCTGGGATCCCAGC CAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTGTTAACATCCATCCTGTGGAGGAGGAG GATGCTGCAACCTATTACTGTCAGCAAAGTACTGAGGATCCTCGGACGTTCGGTGGAGGCACCA AGCTGGAAATCAAACGGGCTGATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCA GTTAACATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAAAGACATCAAT GTCAAGTGGAAGATTGATGGCAGTGAACGACAAAATGGCG (SEQ ID NO: 36; где CDR подчеркнуты и где сигнальная последовательность выделена полужирным шрифтом)

ME TD TILLWVLLLWVPGS TGDIVL T QS PAS LAVS P GQRATIS CKASQSLDYEGDSDMNW

YOQKPGOPPRLLISGASNLESGIPARFSGSGSGTDFTVNIHPVEEEDAATYYCQQSTEDPRTFG

GGTKLEIK

- 20 039293

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38)

CDR-L2: GASNLES (SEQ ID NO: 39)

CDR-L3: QQSTEDPRT (SEQ ID NO: 40)

The parent 22D2 mouse heavy or light chains may be referred to herein as LB145.22D2.E1.1D1. The mouse 19E8 parent heavy or light chains may be referred to herein as LB148.19E8.G1.1A1. The mouse 4A10 parental heavy or light chains may be referred to herein as LB148.4A1O.1H1. The parent murine 11C9 heavy or light chains may be referred to as LB148.11C9.1C1.

The present invention also includes an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., humanized antibodies, such as humanized antagonist antibodies) containing one or more of heavy and/or light chains (or variants) or CDRs (or variants thereof) or mature fragments. such chains (or variants thereof) or variable domains of such chains (or variants thereof) as indicated below. Light chains may be referred to as VL or VK and heavy chains may be referred to as VH.

Mouse immunoglobulin chains.

As such, in the present description there may be mentioned the chains indicated below, having the designation 4A10, 19E8, 11C9 or 22D2. As described herein, the scope of the present invention includes anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments thereof, comprising any one or more (e.g., 3) light chain CDRs and/or any one or several (eg 3) heavy chain CDRs from the immunoglobulin chains listed below; or any mature immunoglobulin light chain variable domain and/or mature immunoglobulin heavy chain variable domain as set forth in SEQ ID NOs: 45-104.

Humanized chains.

In one embodiment, the humanized (eg, humanized antagonist antibody) against LAG3 or antigen-binding fragment of the invention comprises any combination of heavy and light mature variable domains of the following immunoglobulin chains. In one embodiment, the humanized 11C9 light chains are paired with humanized 11C9 heavy chains; humanized 19E8 light chains are paired with humanized 19E8 heavy chains; humanized 4A10 light chains are paired with humanized 4A10 heavy chains; and humanized 22D2 light chains are paired with humanized 22D2 heavy chains. In one embodiment, the humanized anti-LAG3 antibody, or antigen-binding fragment thereof, comprises an immunoglobulin 45AGX 22D2 VL3 light chain variable domain, e.g. ); and an immunoglobulin heavy chain variable domain of Humanizedx[LAG3_H] mAb.22D2 VH6 N54D or Humanizedx[LAG3_H] mAb.22D2 VH6 N54G containing the amino acid sequence of SEQ ID NO: 426 or SEQ ID NO: 427, respectively.

As such, there may be mentioned the circuits indicated below, having the designation 4A10, 19E8, 11C9 or 22D2. As discussed below, the scope of the present invention includes anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antibodies antagonists) and antigen-binding fragments thereof, containing any one or more (e.g., 3) light chain CDRs and/or any one or more (e.g., 3) heavy chain CDRs from the immunoglobulin chains described herein; or any mature or full length VL domain or immunoglobulin light chain and/or mature or full length VH domain or immunoglobulin heavy chain as defined herein. The scope of the present invention also includes any of the humanized polypeptides or polynucleotides, or variable domains thereof, having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations or point deletions.

Specific embodiments of the invention include any humanized anti-LAG3 antibodies and antigen-binding fragments thereof comprising the immunoglobulin light and heavy chains listed below, or any antibody or fragment having their light and heavy chain CDRs (eg, IgG1 or IgG4). Such antibodies and fragments may be referred to as any of Abi, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9 as follows:

Abi: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 53AHH Humanizedx[LAG3_H] mAb (LB145.22D2.E1. D1 VH6) IgGl/Kappa (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

- 21 039293

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLS S PVTKS FNRGEC (аминокислоты 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQS GPEVKKPGT SVKVS CKAS GYT FT DYNVDWVRQARGQRLEWIGDINPNNGGTI

YAQKFQERVTITVDKS Т S TAYMELS S LRS Е DTAVYYCARNYRW FGAMDHWGQGT TVTVS SAS ТК GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK (SEQ ID NO: 106) ;

or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID N0:126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTI

YAQKFQERVTITVDKS T S TAYME L S S LRSE DTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ ID NO: 106 (CDRs underlined)); or containing CDR: CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35)

Ab2: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 56AH Humanizedx[LAG3_H] mAb (LB145.22D2.E1. D1 VH6 N55S) IgG1/Kappa (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLS S PVTKS FNRGEC (amino acids 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQS GPEVKKPGT SVKVSCKAS GYT FT DYNVDWVRQARGQRLEWIGDINPNS GGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK (SEQ ID NO: 108); or an immunoglobulin light chain variable domain containing the amino acid sequence:

- 22 039293

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGOPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCOQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID N0:126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

OMOLVOSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGORLEWIGDINPNSGGTI

YAQKFQERVTITVDKS TS TAYMELS SLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVS S (amino acids 1-119 of SEQ ID NO: 108 (CDR underlined)); or containing a CDR:

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35).

Ab3: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 54AH Humanizedx[LAG3_H] mAb (LB145.22D2.E1. D1 VH6 N55D) IgG1/Kappa (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (аминокислоты 21-238 SEQ ID NO: 126);

an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASТК GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK (SEQ ID NO: 110); or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTOTPLSLSVTPGOPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCOQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID N0:126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

OMOLVOS GPEVKKPGT SVKVS CKASGYT FT DYNVDWVROARGORLEWIGDINPNDGGTI

YAQKFQERVTITVDKS T S TAYMELS S LRSEDTAVYYCARNYRWFGAMDHWGOGTTVTVSS (amino acids 1-119 SEQ ID NO: 110 (CDR underlined)); or containing a CDR:

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35).

Ab4: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3))

Kappa (PX) (or its variable domain) and humanized heavy chain 52AH

Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 VH6 N55Q) IgG1/Kappa (PX) (or its variable to

- 23 039293 men); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (аминокислоты 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence: QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK (SEQ ID NO: 112); or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLOKPGOPPOLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCOQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID N0:126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSTALEGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTI

YAQKFQERVTITVDKS T 3 TAYMEL 3 S LRS EDTAVYYCARNYRWFGAMDHWGOGTTVTVS3 (amino acids 1-119 SEQ ID NO: 112 (CDR underlined)); or containing a CDR:

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35)

Ab5: Humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 57AH Humanized* [LAG3_H] mAb (LB145.22D2.E1 .D1 VH6) IgG4 S228P (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP

PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (amino acids 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTK

GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV

VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLM

ISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG

KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLS LGK (SEQ ID NO: 114); or

- 24 039293 immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKRGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDR underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGORLEWIGDINPNNGGTI

YAQKFQERVTITVDKS T 3 TAYME L S S LRS E DTAVYYCARNYRW FGAMDHWGOG T T T UT VS S (amino acids 1-119 SEQ ID NO: 114 (CDR underlined)); or containing a CDR:

CDR-L1: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35).

Ab6: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3))

Kappa (PX) (or its variable domain) and humanized heavy chain 73AHD Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 VH6 N55D/VL3) IgG4 S228P/Kappa (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP

PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLS S PVTKS FNRGE C (amino acids 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTI

YAQKFQERVTITVDKS TS TAYMELS 3 LRSEDTAVYYCARNYRWFGAMDHWGQGT TVTVS SAS TK

GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV

VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLM

ISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG

KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS

LGK (SEQ ID NO: 116) ; or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTOTPLSLSVTPGOPASISCKASQSLDYEGDSDMNWYLQKPGOPPOLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDR underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

OMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTI

YAQKFQERVTITVDKS T S TAYME L S S LRSEDTAVYYCARNYRWFGAMDHWGOGTTVTVSS (amino acids 1-119 SEQ ID NO: 116 (CDR underlined)); or containing a CDR:

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35).

Ab7: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3))

Kappa (PX) (or its variable domain) and humanized heavy chain 21AHG

- 25 039293

Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 VH6 N55S/VL3) IgG4 S228P/Kappa (PX) (or variable domain thereof); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (amino acids 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTI

YAQKFQERVTITVDKS T S TAYMELS S LRSEDTAVYYCARNYRW FGAMDHWGQGT TVTVS SAS TK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLM

ISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQSEQLS8 (ID: NO: LGK 1); or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCOQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

QMQLVQSGPEVKKPGТSVKVSCKASGYТFTDYNVDWVRQARGQRLEWIGDINPNSGGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGOGTTVTVSS (amino acids 1-119 SEQ ID NO: 118 (CDR underlined)); or containing a CDR:

CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35).

Ab8: humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 80AHG Humanizedx[LAG3_H] mAb (LB145.22D2.E1. D1 VH6 N55Q/VL3) IgG4 S228P/Kappa (PX) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP

PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLS S PVTKS FNRGE C (amino acids 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

- 26 039293

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTI YAQKFQERVTITVDKS T S TAYME LSSLRSED TAVYYCARNYRW FGAMDHWGQGT TVTVSSAS TK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLM ISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO: 120); or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTOTPLSLSVTPGOPASISCKASQSLDYEGDSDMNWYLQKPGOPPOLLIYGASNLE

SGVPDRFSGSGSGТDFTLKISRVEАEDVGVYYCQQSTEDPRTFGGGТKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSOKASGYTFTDYNVDWVRQARGORLEWIGDINPNQGGTI

YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGOGTTVTVSS (amino acids 1-119 SEQ ID NO: 120 (CDR underlined)); or containing a CDR:

CDR-L1: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35);

or

Ab9: Humanized light chain 45AGX Humanizedx[LAG3_H] mAb (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or variable domain thereof) and humanized heavy chain 72AHD Humanized* [LAG3_H] mAb (LB145.22D2.E1 .D1 VH6 N55G/VL3) IgG4 S228P/Kappa (PX)) (or its variable domain); for example, containing an immunoglobulin light chain containing the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLE SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLS S PVTKS FNRGEC (аминокислоты 21-238 SEQ ID NO: 126); and an immunoglobulin heavy chain containing the amino acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNGGGTI YAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLM ISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO: 122); or an immunoglobulin light chain variable domain containing the amino acid sequence:

DIVMTOTPLSLSVTPGOPASISCKASQSLDYEGDSDMNWYLOKPGOPPOLLIYGASNLE

SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDR underlined)); and an immunoglobulin heavy chain variable domain containing the amino acid sequence:

- 27 039293

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNGGGTI

YAQKFQERVTITVDKS TS TAYMELS SLRSEDTAVYYCARNYRWFGAMDHWGQGT TVTVS S (amino acids 1-119 of SEQ ID NO: 122 (CDR underlined)); or containing CDR: CDR-Ll: KASQSLDYEGDSDMN (SEQ ID NO: 38);

CDR-L2: GASNLES (SEQ ID NO: 39);

CDR-L3: QQSTEDPRT (SEQ ID NO: 40);

CDR-H1: DYNVD (SEQ ID NO: 33);

CDR-H2: DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and

CDR-H3: NYRWFGAMDH (SEQ ID NO: 35)

In one embodiment, the CDR-H2 of any anti-LAG3 antibody or antigen-binding fragment thereof of the present invention contains the amino acid sequence:

DINPNXiGGTIYX ₂ QKFX ₃ X ₄ (SEQ ID NO: 446) where Xi=D, N, S or Q;

X ₂ =A or S;

X ₃ Q or K;

X ₄ \u003d E or G.

Humanized immunoglobulin heavy chains are listed in

SEQ ID NO: 106, 108, 110, 112, 114, 116, 118, 120

124, 128, 134

160, 162, 164,

186, 188, 190,

216, 218, 220,

253, 255, 265

287, 289, 291,

313, 315, 317,

339, 341, 343,

367, 377, 379,

140, 142,

166, 168,

192, 194,

222, 234,

267, 269,

293, 295,

319, 321,

345, 347

381, 383,

144, 146

170, 172,

196, 198,

235, 237,

271, 273,

297, 299

323, 325,

349, 353,

385, 387,

148, 150,

174, 176,

200, 202,

239, 243,

275, 277

301, 303,

327, 329

355, 357,

389, 391,

152, 154

178, 180,

204, 206,

245, 247,

279, 281,

305, 307,

331, 333,

359, 361,

393, 395,

122, 124

156, 158,

182, 184,

212, 214,

249, 251,

283, 285,

309, 311,

335, 337

363, 365,

397, 399,

406-419, 448, 449, 462 and 463.

DNA encoding humanized heavy chains are listed in

SEQ ID NO: 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 127, 133, 139, 141, 143, 145, 147, 149,151,

153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175,177,

179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201,203,

205, 211, 213, 215, 217, 219, 221, 233, 236, 238, 242, 244,246,

248, 250, 252, 254, 264, 266, 268, 270, 272, 274, 276, 278,280,

282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304,306,

308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330,332,

334, 336, 338, 340, 342, 344, 346, 348, 352, 354, 356, 358,360,

362, 364, 366, 376, 378, 380, 382, 384, 386, 388, 390, 392,394,

396 and 398.

Humanized immunoglobulin light chains are listed in

SEQ ID NO: 126, 130, 132, 136, 138, 208, 210, 224, 226, 228, 230,

232, 241, 257, 259, 261, 263, 351, 369, 371, 373, 375, 401, 403,

405, 450-453, 426, 427 and 459-461.

DNAs encoding humanized immunoglobulin light chains are listed in

SEQ ID NOs: 125, 129, 131, 135, 137, 207, 209, 223, 225, 227, 229, 231,

240, 256, 258, 260, 262, 350, 368, 370, 372, 374, 400, 402 and 404.

A variant polypeptide, such as an immunoglobulin chain, refers to a polypeptide containing an amino acid sequence that is at least about 70-99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83 , 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) is identical or similar to the mentioned amino acid sequence, referred to in this

-28039293 describing when a comparison is made using the BLAST algorithm, where the algorithm parameters are chosen to achieve the best match between matching sequences over the entire length of the matching reference sequences (e.g., expected threshold: 10; word length: 3; maximum number of matches in the requested range : 0; BLOSUM matrix 62; gap penalty: insertion 11, continuation 1; conditional correction of the compositional evaluation matrix).

A variant polynucleotide refers to a polynucleotide containing a nucleotide sequence that is at least about 70-99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, , 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) is identical to the mentioned nucleotide sequence, which is specified in the present description, when the comparison is carried out using the BLAST algorithm, where the algorithm parameters are chosen to achieve the best match between matching sequences over the entire length of the matching reference sequences (e.g., expected threshold: 10; word length: 28; maximum number of matches in the requested range: 0; pass/fail score : 1, -2; gap penalty: linear).

In one embodiment, the anti-LAG3 polypeptides and antibodies and antigen-binding fragments thereof (e.g., humanized antibodies) of the present invention comprise an immunoglobulin heavy chain variable region having at least 78.99% (e.g., 79, 80, 85, 90, 95, 99%) amino acid sequence identity with amino acids 1-119 of SEQ ID NO: 106; and/or an immunoglobulin light chain variable region having at least 78.38% (e.g., 79, 80, 85, 90, 95, 99%) amino acid sequence identity with amino acids 1-111 of SEQ ID NO: 224.

In addition, a variant may be a polypeptide containing the amino acid sequence as defined herein, excluding one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) mutations, e.g. such as missense mutations (eg conservative substitutions), nonsense mutations, deletions or insertions. Such a polypeptide may be an immunoglobulin light chain, an immunoglobulin heavy chain, and/or a CDR (e.g., any one or more of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and/or CDR-H3) .

As described herein, the present invention relates to anti-LAG3 antibodies and antigen-binding fragments thereof, which comprise one or more variant framework sequences (e.g., any one or more of FR-L1, FR-L2, FR-L3, FR-L4, FR-H1, FR-H2, FR-H3 and/or FR-H4), CDRs (e.g. 1, 2 or 3 CDR-L variants and/or 1, 2 or 3 CDR-H variants) and/or immunoglobulin chains (for example, 1 or 2 variants of V _L and/or 1 or 2 variants of VH), the sequences of which are expressly indicated in the present description. Such antibodies and antigen-binding fragments may themselves be referred to as variants. Simple polypeptide chains that include one or more variants of FR, CDR-L, CDR-H and/or immunoglobulin chains are themselves part of the present invention. Also part of the present invention are polynucleotides encoding such variants of the polypeptide portions. For example, the present invention relates to antibodies against LAG3 and antigen-binding fragments thereof, which contain the amino acid sequence V _H and V _L of antibodies Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9, as described in the present description, as well as their variants containing the CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of said Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9, but having 70% or more (e.g., 80, 85, 90, 95, 97, or 99%) common amino acid sequence identity or similarity to the indicated VH and V _L Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, or Ab9. Thus, in such embodiments, the CDRs of the antibodies and fragments are identical to the VH and V _L of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, or Ab9, however, any differences from such VHs occur in the framework regions and/or constant domains of the immunoglobulins. and V.L.

The invention relates to anti-LAG3 antibody variants or antigen-binding fragments thereof (e.g., humanized antibodies, such as humanized antagonist antibodies) containing one or more CDR variants (e.g., any one or more of CDR-L1, CDRL2, CDR-L3, CDR -H1, CDR-H2 and/or CDR-H3) and/or framework regions (eg any one or more of FR1, FR2, FR3 and/or FR4) as described herein; and/or one or more variants of VL domains and/or one or more variants of V _H domains of such antibodies or fragments as specified in the present description, for example, having at least 70, 75, 80, 85, 90, 95, 98, 99, or 99.9% sequence identity or similarity, e.g. to

- 29 039293

SEQ ID NO: 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14,15,

17, 18, 19, 20, 22, 23, 24, 25, 27, 28, 29, 30, 32, 33, 34.35,

37, 38, 39, 40, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65.67,

69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97.99,

101, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124,126,

128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150,152,

154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176.178,

180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202,204,

206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228,230,

232, 234, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253.255,

257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279.281,

283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307,

309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331,333,

335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357,359,

361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383,385,

387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 406, 407,408,

409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 426,427,

434, 435, 436, 437, 438, 439, 440, 441, 442, 446, 448, 449,451,

452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463 or 464;

which specifically bind to LAG3.

As discussed above, the scope of the present invention includes anti-LAG3 antibody variants or antigen-binding fragments containing one or more CDR variants (eg, 1, 2, or 3 CDR-L variants and/or 1, 2, or 3 CDR-H variants); and/or framework regions (e.g. any one or more of FR1, FR2, FR3 and/or FR4) and/or VL domain variants and/or VH domain variants (with or without signal sequence) having 1, 2, 3 , 4, 5, 6, 7, 8, 9, 10 or more mutations. Mutations may include point mutations, which are conservative or non-conservative amino acid substitutions or point deletions, for example, in the framework region and/or in the CDR. As discussed above, the present invention relates to antibodies against LAG3 and antigen-binding fragments thereof, containing CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDRH2 and CDR-H3 of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8 or Ab9, but having mutations in their framework regions.

Conservatively modified anti-LAG3 antibody variants and antigen-binding fragments thereof are also part of the present invention. A conservatively modified variant or conservative substitution refers to a variant that has one or more amino acid substitutions for the polypeptide with other amino acids that have similar characteristics (e.g., charge, side chain size, hydrophobicity/hydrophilicity, backbone conformation and rigidity, etc.). Such changes can often be made without significantly disrupting the biological activity of the antibody or fragment. Those skilled in the art will appreciate that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin/Cummings Pub. Co., p.224 (4th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to interfere with biological activity.

Variants of antibodies against LAG3 or antigennegative fragments of the present invention, which are described in the present description, contain one or more CDR (for example, 1, 2 or 3 variants of CDR-L and/or 1, 2, or 3 variants of CDR-H); framework regions (eg, any one or more of FR1, FR2, FR3 and/or FR4); and/or immunoglobulin heavy chains having one or more conservative substitutions. For example, such antibodies and fragments may contain the amino acid sequences described herein, e.g.

- 30 039293

SEQ ID NO: 2, 3, 4, 5, 7, 8, 9, 10, 12, 13,14,

22, 23, 24, 25, 27, 28, 29, 30, 32, 33.34,

45, 47, 49, 51, 53, 55, 57, 59, 61, 63.65,

77, 79, 81, 83, 85, 87, 89, 91, 93, 95.97,

108, 110, 112, 114, 116, 118, 120, 122,124,

134, 136, 138, 140, 142, 144, 146, 148,150,

160, 162, 164, 166, 168, 170, 172, 174.176,

186, 188, 190, 192, 194, 196, 198, 200,202,

212, 214, 216, 218, 220, 222, 224, 226,228,

237, 239, 241, 243, 245, 247, 249, 251,253,

263, 265, 267, 269, 271, 273, 275, 277.279,

289, 291, 293, 295, 297, 299, 301, 303,305,

315, 317, 319, 321, 323, 325, 327, 329.331,

341, 343, 345, 347, 349, 351, 353, 355,357,

367, 369, 371, 373, 375, 377, 379, 381,383,

393, 395, 397, 399, 401, 403, 405, 406,407,

412, 413, 414, 415, 416, 417, 418, 419.426,

437, 438, 439, 440, 441, 442, 446, 448,449,

455, 456, 457, 458, 459, 460, 461, 462, 463

15, 17, 18, 19.20,

35, 37, 38, 39.40,

67, 69, 71, 73.75,

99, 101, 103, 106,

126, 128, 130, 132,

152, 154, 156, 158,

178, 180, 182, 184,

204, 206, 208, 210,

230, 232, 234, 235,

255, 257, 259, 261,

281, 283, 285, 287,

307, 309, 311, 313,

333, 335, 337, 339,

359, 361, 363, 365,

385, 387, 389, 391,

408, 409, 410, 411,

427, 434, 435, 436,

451, 452, 453, 454, or 464;

where such amino acid sequences may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20 or more of their conservative amino acid substitutions. Illustrative conservative substitutions are listed in table. one.

Table 1 Exemplary Conservative Amino Acid Substitutions

original balance conservative replacement Ala (A) Gly; Ser Arg(R) Lys; His Asn(N) gin; His Asp(D) Glu; Asn Cys(C) Ser; Ala Gin (Q) Asn Glu(E) asp; gin Gly (G) Ala His(H) asn; gin He(I) Leu; Vai Leu(L) He; Vai Lys (K) Arg; His Met(M) Leu; He; Tyr Phe(F) Tyr; met; Leu Pro (P) Ala Ser(S) Thr Thr(T) Ser TRP(W) Tyr; Phe Tyr (Y) trp; Phe Wai (V) He; Leu

Functionally conserved anti-LAG3 antibody variants and antigen-binding fragments thereof are also part of the present invention. Any of the variants of anti-LAG3 antibodies and antigen-binding fragments thereof (as described herein) may be functionally conserved variants. In some cases, such functionally conservative variants may also be characterized as conservatively modified variants. Functionally conservative variants, as used herein, refer to variants of anti-LAG3 antibodies or antigen-binding fragments thereof, in which one or more amino acid residues (e.g., 1, 2, 3, 4, 5, or 6 CDRs, and/or V _L , and/or VH) are changed without significant change in one or more functional properties of the antibody or fragment. In one embodiment, the functionally conserved variant of an anti-LAG3 antibody and its antigen-binding fragments of the present invention (e.g., humanized antibodies, such as humanized antagonist antibodies) comprise an immunoglobulin chain variant (e.g., one

- 31 039293 or two VH variants and/or one or more V _L variants) and/or CDRs (for example, 1, 2 or 3 CDR-L variants and/or 1, 2 or 3 CDR-H variants) of any of the immunoglobulin chains and/or CDRs specified in the present description, for example, any of

SEQ ID NO: 2, : 3, 4, 5, 7, 8, 9, 10 , 12, , 13, 14, 15, 17 , eighteen, . 19 twenty, 22, 22 J.24 .25 27, 28, 29, 30 , 32 33, 34, 35, 37 .38 39 40, 45, 47, 49 .51 53, 55, 57, 59 .61 63, 65, 67, 69 .71 . 73 75, 77, 79, 81 83 85 87 89, 91, 93, 95, , 97, 99, 101, 103 106 108 110 112 114 , 116 , 118, 120 122, 124, 126 128 130 132, 134, 136 138, 140, , 142, , 144, 146 148, 150, 152, 154, 156 158, 160, 162, 164, 166, , 168, . 170 172, 174, 176, 178, 180, 182 184, 186, 188, 190, 192, , 194, , 196 198, 200, 202, 204, 206, 208 210, 212, 214, 216, 218, . 220 , 222, 224, 226, 228, 230, 232, 234 235, 237, 239, 241, 243, . 245 . 247 249, 251, 253, 255, 257, 259 261, 263, 265, 267, 269, , 271 . 273 275, 277, 279, 281, 283, 285 287, 289, 291, 293, 295, , 297 , 299 301, 303, 305, 307, 309, 311 313, 315, 317, 319, 321, , 323, , 325, 327, 329, 331, 333, 335, 337 339, 341, 343, 345, 347, , 349, , 351 353, 355, 357, 359 361, 363 365, 367, 369, 371, 373, .375 . 377 379, 381, 383, 385, 387, 389 391, 393, 395, 397, 399 , 401, , 403, 405, 406, 407, 408, 409, 410 411, 412, 413, 414, 415, .416 .417 418, 419, 426, 427, 434, 435 436, 437, 438, 439, 440, .441 442 446, 448, 449, 451, 452, 453 454, 455, 456, .457 .458 459, 460, 461, 462, 463 or 464

exhibiting one or more of the following functional properties:

inhibits binding of LAG3 to MHC class II molecules, for example, on Daudi cells; e.g. inhibits human LAG3/MHC class II binding on Daudi cells with an IC _{50 of} approximately 1.9-2.9 nM e.g. 2.1, 2.8, 2.0, 1.9, 2.5, 2.6 , 2.1, 2.4, or 2.5 nM (eg, about 2.1-2.6 nM).

competes with MHC class II molecules for LAG3 binding, eg on Daudi cells;

connects an additional loop of LAG3;

binds LAG3 with a KD affinity of about 10 ^-9 M to about 2x10 - ¹² M (eg as measured by surface plasmon resonance or KinExA); for example, binds human LAG3 to a KD of about 2, 3, 6, 10, or 11 pM (e.g., 2-11 pM) and/or binds cynomolgus monkey LAG3 to a KD of about 11, 12, 16, or 25 pM (e.g., 11-25 pM). pM), for example, through KinExA;

binds to native LAG3 on the surface of activated CD4+ and/or CD8+ T cells; for example, binds human CD4+ T cells expressing human LAG3 with an EC _{50 of} about 39, 41, or 57 pM (eg, about 39-57 pM); binds human CD8+ T cells expressing human LAG3 with an EC _{50 of} about 33, 35, or 49 pM (eg, about 33-49 pM); binds cynomolgus monkey CD4+ T cells expressing cynomolgus monkey LAG3 to an EC _{50 of} about 27, 30, or 35 pM (eg, about 27-35 pM); binds cynomolgus monkey CD8+ T cells expressing cynomolgus monkey LAG3 to an EC _{50 of} about 30, 31, or 41 pM (eg, about 30-41 pM); for example, where T cells are isolated from blood;

binds to human and/or cynomolgus monkey LAG3, eg Masaca fascicularis or Macaca mulatta;

inhibits LAG3 homodimerization;

stimulates production of IL-2 by antigen-specific T cells, _e.g. 4.06 nM, 2.83-2.96 nM, 0.57-1.07 nM, 0.45-1.27 nM, 0.47-1.01 nM or 0.72-1.08 nM;

carries out labeling of tissue of the tonsil; and/or enhances T cell activation by an anti-PD1 antibody such as pembrolizumab, for example, increases T cell production of IL-2;

does not label brain, heart, kidney, liver, lung, pancreas and/or pituitary tissue;

binds to human LAG3 via contact between residues Q ^E G ^APA Q ^L (amino acids 35-42 SEQ ID NO: 443) and RPARRADAGEYRAAVH (amino acids 137-152 SEQ ID NO: 443) and optionally residues DERGRQRGDFSLW (amino acids 123-135 SEQ ID NO: 443) LAG3; or residues SPTIPLQDL (amino acids 45-53 SEQ ID NO: 443) and optionally dergrqrgdfsl (amino acids 123-134 SEQ ID NO: 443) LAG3; or residues hplapgphpaapsswgprprrytvl (amino acids 78-102 SEQ ID NO: 443) LAG3; and/or by protecting the hydrogen atoms on the amide backbone of such residues from being replaced by deuterium (eg from D ₂ O).

- 32 039293

The present invention relates to a method for producing an antibody, or antigen-binding fragment thereof, that specifically binds to LAG3, comprising administering to a non-human host animal (e.g., mouse, rabbit, camel, llama, or rat) an effective amount of one or more peptides containing, consisting of or essentially consisting of an amino acid sequence selected from the group consisting of

QEGAPAQL (amino acids 35-42 SEQ ID NO: 443);

RPARRADAGEYRAAVH (amino acids 137-152 SEQ ID NO: 443);

DERGRQRGDFSLW (amino acids 123-135 SEQ ID NO: 443);

SPTIPLQDL (amino acids 45-53 of SEQ ID NO: 443);

dergrqrgdfsl (amino acids 123-134 SEQ ID NO: 443); and

HPLAPGPHPAAPSSWGPRPRRYTVL (amino acids 78-102 SEQ ID NO: 443), for example, where the peptide is formulated with a pharmaceutically acceptable carrier. Optionally, the antibody or fragment is isolated from the host animal, for example, from the serum or blood of the host animal. Optionally, more than one dose of the peptide is administered to the host animal. Such isolated peptides are part of the present invention, for example, when fused to an immunogen such as keyhole limpet hemocyanin (KLH), human serum albumin or bovine serum albumin.

Sequence identity refers to the degree to which the amino acids of two polypeptides are the same at equivalent positions when the two sequences are optimally aligned. Sequence similarity includes identical residues and non-identical, biochemically related amino acids. Biochemically related amino acids that have similar properties and can be interchanged are discussed above.

The following links refer to BLAST algorithms often used for sequence analysis:

BLAST algorithms: Altschul et al. (2005) FEBS J. 272(20): 5101-5109; Altschul, S.F., et al. (1990) J. Mol. Biol. 215:403-410; Gish, W., et al. (1993) Nature Genet. 3:266-272; Madden, T.L., et al. (1996) Meth. Enzymol. 266:131-141; Altschul, S.F., et al. (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J., et al. (1997) Genome Res. 7:649-656; Wootton, J.C., et al. (1993) Comput. Chem. 17:149-163; Hancock, J.M. et al. (1994) Comput. Appl. biosci. 10:67-70;

alignment scoring systems: Dayhoff, M.O., et al., A model of evolutionary change in proteins, Atlas of Protein Sequence and Structure, (1978) vol. 5 suppl. 3.M.O. Dayhoff (ed.), p. 345-352, Natl. Biomed. Res. Found., Washington, DC; Schwartz, R.M., et al., Matrices for detecting distant relationships, Atlas of Protein Sequence and Structure, (1978) vol. 5 suppl. 3.M.O. Dayhoff (ed.), p. 353-358, Natl. Biomed. Res. Found., Washington, DC; Altschul, S.F. (1991) J. Mol. Biol. 219:555-565; States, D.J., et al. (1991) Methods 3:66-70; Henikoff, S., et al. (1992) Proc. Natl. Acad. sci. USA 89:10915-10919; Altschul, S.F., et al. (1993) J. Mol. Evol. 36:290-300;

alignment statistics: Karlin, S., et al. (1990) Proc. Natl. Acad. sci. USA 87:2264-2268; Karlin, S., et al. (1993) Proc. Natl. Acad. sci. USA 90:5873-5877; Dembo, A., et al. (1994) Ann. Prob. 22:2022-2039; and Altschul, S.F. Evaluating the statistical significance of multiple distinct local alignments, Theoretical and Computational Methods in Genome Research (S. Suhai, ed.), (1997) p. 1-14, Plenum, New York.

The anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention (e.g., humanized antibodies, such as humanized antagonist antibodies) may contain one, two, three, four, five, or six of the complementarity determining regions (CDRs) of the immunoglobulin chains described in the present invention. (where 1, 2, 3, 4, 5 or 6 of the CDRs are optionally variants of the CDRs described in the present invention). One, two, three, four, five or six CDRs can be independently selected from the CDR sequences of the various immunoglobulin chains described in the present invention. Alternatively, one, two, three, four, five, or six CDRs may be selected from the CDR sequences of one described antibody of the invention.

For example, the present invention provides anti-LAG3 antibodies and antigen-binding fragments thereof, as well as immunoglobulin polypeptide chains, which contain:

CDR-H1, CDR-H2 and CDR-H3 4A10;

CDR-L1, CDR-L2 and CDR-L3 4A10;

CDR-H1, CDR-H2 and CDR-H3 11C9;

CDR-L1, CDR-L2 and CDR-L3 11C9;

CDR-H1, CDR-H2 and CDR-H3 19E8;

CDR-L1, CDR-L2 and CDR-L3 19E8;

CDR-H1, CDR-H2 and CDR-H3 22D2 and/or

CDR-L1, CDR-L2 and CDR-L3 22D2;

where CDRs 4A10, 11C9, 19E8 and 22D2 may be derived from mouse immunoglobulin chains or humanized immunoglobulin chains 4A10, 11C9, 19E8 and 22D2, respectively, specified in this

- 33 039293 in the general description (for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9).

Anti-LAG3 antibodies, or antigen-binding fragments thereof, of the present invention may comprise at least one antibody heavy chain variable domain (VH) comprising one or more (e.g., 3) of CDR-H1, CDR-H2, or CDR-H3 VH 4A10 (e.g., , SEQ ID NO: 2); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 3 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 4 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), and 5 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention may comprise at least one antibody light chain variable domain (VL) comprising one or more (e.g., 3) of CDR-L1, CDR-L2, and CDR-L3 V _L 4A10 ( for example, SEQ ID NO: 7); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 8 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 9 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 10 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

Anti-LAG3 antibodies, or antigen-binding fragments thereof, of the present invention may comprise at least one heavy chain variable domain (V _H ) of an antibody comprising one or more (e.g., 3) of CDR-H1, CDR-H2, or CDR-H3 V _H 19E8 (for example, SEQ ID NO: 12); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 13 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 14 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 15 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

Anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention may comprise at least one light chain variable domain (VL) of an antibody comprising one or more (e.g., 3) of CDR-L1, CDR-L2, and CDR-L3 V _L 19E8 ( for example, SEQ ID NO: 17); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 18 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations and/or point deletions), 19 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 20 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

Anti-LAG3 antibodies, or antigen-binding fragments thereof, of the present invention may comprise at least one antibody heavy chain variable domain (VH) comprising one or more (e.g., 3) of CDR-H1, CDR-H2, or CDR-H3 VH 11C9 (e.g., , SEQ ID NO: 22); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 23 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations and/or point deletions), 24 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 25 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention may comprise at least one antibody light chain variable domain (V _L ) comprising one or more (e.g., 3) of CDR-L1, CDR-L2, and CDR-L3 V _L 11C9 (for example, SEQ ID NO: 27); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 28 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations and/or point deletions), 29 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 30 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention may comprise at least one antibody heavy chain variable domain (VH) comprising one or more (e.g., 3) of CDR-H1, CDR-H2, or CDR-H3 V _H 22D2 ( for example, SEQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118, 120 or 122); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 33 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations and/or point deletions), 34 , 446, 454, 455, 456, 457, or 458 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 point mutations and/or point deletions) and 35 (or its a variant having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

Anti-LAG3 antibodies, or antigen-binding fragments thereof, of the present invention may comprise at least one light chain variable domain (VL) of an antibody comprising one or more (e.g., 3) of CDR-L1, CDR-L2, and CDR-L3 V _L 22D2 ( for example, SEQ ID NO: 37 or 126); for example, where the CDRs contain the amino acid sequences shown in SEQ ID NO: 38 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 39 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 40 (or a variant thereof having 1, 2, 3, 4, 5 , 6, 7, 8, 9 or 10 point mutations and/or point deletions), respectively.

The present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof.

- 34 039293 mentu, which contain:

CDR-H1, CDR-H2 and CDR-H3 4A10; and CDR-L1, CDR-L2 and CDR-L3 4A10;

CDR-H1, CDR-H2 and CDR-H3 11C9; and CDR-L1, CDR-L2 and CDR-L3 11C9;

CDR-H1, CDR-H2 and CDR-H3 19E8; and CDR-L1, CDR-L2 and CDR-L3 19E8; or

CDR-H1, CDR-H2 and CDR-H3 22D2; and CDR-L1, CDR-L2 and CDR-L3 22D2;

where CDRs 4A10, 11C9, 19E8 and 22D2 may be derived from murine immunoglobulin chains or humanized immunoglobulin chains 4A10, 11C9, 19E8 and 22D2, respectively, as described herein, and where optionally 1, 2, 3, 4, 5, or 6 of the CDRs are variants of the CDRs referred to herein.

The present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof, which comprise a light chain variable domain (V _L ) containing CDR-L1, CDR-L2 and CDRL3 V _L 4A10 (e.g., SEQ ID NO: 8 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 9 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8 , 9 or 10 point mutations and/or point deletions) and 10 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)); and a heavy chain variable domain (VH) of an antibody comprising CDR-H1, CDR-H2, and CDR-H3 V _H 4A10 (e.g., SEQ ID NO: 3 (or a variant thereof having 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 point mutations and/or point deletions), 4 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions ) and 5 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, comprises an antibody light chain variable domain (VL) comprising CDR-L1, CDR-L2, and CDR-L3 V _L 19E8 (e.g., SEQ ID NO: 18 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 19 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8 , 9 or 10 point mutations and/or point deletions) and 20 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)); and a heavy chain variable domain (VH) of an antibody containing CDR-H1, CDR-H2, and CDR-H3 V _H 19e8 (e.g., SEQ ID NO: 13 (or a variant thereof having 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 point mutations and/or point deletions), 14 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions ) and 15 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, comprises the light chain variable domain (V _L ) of the antibody comprising CDR-L1, CDR-L2, and CDR-L3 V _L 11C9 (e.g., SEQ ID NO: 28 (or a variant thereof, having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 29 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 30 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)); and a heavy chain variable domain (VH) of an antibody comprising CDR-H1, CDR-H2, and CDR-H3 11C9 VH (e.g., SEQ ID NO: 23 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 24 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 25 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, the anti-LAG3 antibody, or antigen-binding fragment thereof, comprises the light chain variable domain (V _L ) of the antibody comprising CDR-L1, CDR-L2, and CDR-L3 V _L 22D2 (e.g., SEQ ID NO: 38 (or a variant thereof, having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 39 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 40 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)); and a heavy chain variable domain (VH) of an antibody comprising CDR-H1, CDR-H2, and CDR-H3 V _H 22d2 (e.g., SEQ ID NO: 33 (or a variant thereof having 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 point mutations and/or point deletions), 34, 446, 454, 455, 456, 457 or 458 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 35 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, the antibody is a humanized LAG3 antagonist antibody. Examples of such humanized anti-LAG3 antibodies include, but are not limited to, antibodies containing CDR-L1, CDR-L2 and CDR-L3 4A10; and CDR-H1, CDR-H2 and CDRH3 4A10.

In a further embodiment, the antibody is a humanized LAG3 antagonist antibody. Examples of such humanized anti-LAG3 antibodies include, but are not limited to, antibodies containing CDR-L1, CDR-L2 and CDR-L3 19E8; and CDR-H1, CDR-H2 and CDRH3 19E8.

In a further embodiment, the antibody is a humanized LAG3 antagonist antibody. Examples of such humanized anti-LAG3 antibodies include, but are not limited to, antibodies containing CDR-L1, CDR-L2 and CDR-L3 11C9; and CDR-H1, CDR-H2 and CDRH3 11C9.

In the following embodiment, the antibody is a humanized antibody

- 35 039293 antagonist against LAG3. Examples of such humanized anti-LAG3 antibodies include, but are not limited to, antibodies containing CDR-L1, CDR-L2 and CDR-L3 22D2; and CDR-H1, CDR-H2 and CDRH3 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9.

The present invention relates to an anti-LAG3 antibody, or an antigen-binding fragment thereof, or an immunoglobulin polypeptide, which comprises:

mature domain V _L immunoglobulin 4A10 and/or mature domain V _H 4A10;

mature V _L domain of immunoglobulin 19E8 and/or mature VH domain 19E8;

11C9 immunoglobulin V _L mature domain and/or 11C9 VH mature domain; and/or a mature 22D2 immunoglobulin V _L domain and/or a mature 22D2 VH domain;

where the V _L or VH domain of 4A10, 19E8, 11C9 and 22D2 is a mouse V _L or VH domain or a humanized V _L or VH domain of 4A10, 19E8, 11C9 and 22D2 described herein, and where optionally V _L and/or VH are a V _L or VH variant as described herein (eg, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, or Ab9).

In addition, the present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof, which contain the mature V _L domain 4A10, 19E8, 11C9 or 22D2, where the V _L domain contains the amino acid sequence

SEQ ID NO: 7, 17, 27

37, 57, 59, 61, 63, 65, 101, 126, 130, 132, 136, 138, 208, 210,

224, 226, 228, 230, 232, 241, 257, 259, 261, 263, 351, 369, 371,

373, 375, 401, 403, 405, 426, 427, 450-453, or 459-461, or a variant thereof.

In addition, the present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof, which contain the mature VH domain 4A10, 19E8, 11C9 or 22D2, where the VH domain contains the amino acid sequence

SEQ ID NO: 2, 12, 22

32, 45, 47, 49, 51, 53, 55, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 128, 134, 140, 142, 144, 146, 148,150,

152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174,176,

178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200.202,

204, 206, 212, 214, 216, 218, 220, 222, 234, 235, 237, 239.243,

245, 247, 249, 251, 253, 255, 265, 267, 269, 271, 273, 275,277,

279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301,303,

305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327,329,

331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 353, 355,357,

359, 361, 363, 365, 367, 377, 379, 381, 383, 385, 387, 389,391,

393, 395, 397, 399, 406-419, 434-442, 448, 449, 462, 463 or 464;

or their version.

In addition, the present invention relates to an anti-LAG3 antibody, or antigen-binding fragment thereof, which contains the mature V _L domain of 4A10 (e.g., amino acids 21-132 of SEQ ID NO: 7 or a variant thereof) and the mature VH domain of 4A10 (e.g., amino acids 20 -136 SEQ ID NO: 2 or variants thereof).

In addition, the present invention provides an anti-LAG3 antibody, or antigen-binding fragment thereof, that comprises the mature V _L domain of 19E8 (e.g., amino acids 21-128 of SEQ ID NO: 17 or a variant thereof) and the mature VH domain of 19E8 (e.g., amino acids 20 -138 SEQ ID NO: 12 or variant).

In addition, the present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof, which contain a mature 11C9 V _L domain (e.g., amino acids 21-128 of SEQ ID NO: 27 or a variant thereof) and a mature 11C9 VH domain (e.g., amino acids 20 -142 SEQ ID NO: 22 or variant).

In addition, the present invention relates to an anti-LAG3 antibody or antigen-binding fragment thereof, which contain a mature V _L domain of 22D2 (for example, amino acids 21-131 of SEQ ID NO: 37 or 126 or a variant thereof) and a mature VH domain of 22D2 (for example, amino acids 21-138 or 21-131 of SEQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118, 120, or 122 or a variant thereof).

The invention also relates to polypeptides containing the amino acid sequence specified in SEQ ID NO: 2, or its amino acids 20-136, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence,

- 36 039293 specified in SEQ ID NO: 7, or amino acids 21-132 thereof, or a variant thereof; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence specified in SEQ ID NO: 12, or its amino acids 20-138, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence specified in SEQ ID NO: 17, or its amino acids 21-128, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence specified in SEQ ID NO: 22, or its amino acids 20-142, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence specified in SEQ ID NO: 27, or its amino acids 21-128, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence indicated in SeQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118, 120 or 122, or its mature fragment, for example, containing its amino acids 20-138 or 20-131; or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides containing the amino acid sequence indicated in SEQ ID NO: 37 or 126, or its Mature fragment, for example, containing its amino acids 21-131, or its variant; or to any polynucleotide encoding such a polypeptide.

The invention also relates to polypeptides (eg, humanized immunoglobulin chain) containing CDR-H1, CDR-H2 and CDR-H3 V _H -domain containing SEQ ID NO: 2 (eg, SEQ ID NO: 3-5); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, a humanized immunoglobulin chain) containing CDR-L1, CDR-L2 and CDR-L3 of a VL domain containing SEQ ID NO: 7 (eg, SEQ ID NO: 8-10); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, humanized immunoglobulin chain) containing CDR-H1, CDR-H2 and CDR-H3 V _H -domain containing SEQ ID NO: 12 (eg, SEQ ID NO: 13-15); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, humanized immunoglobulin chain) containing CDR-L1, CDR-L2, and CDR-L3 VL domain containing SEQ ID NO: 17 (eg, SEQ ID NO: 18-20); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, humanized immunoglobulin chain) containing CDR-H1, CDR-H2 and CDR-H3 V _H -domain containing SeQ ID NO: 22 (eg, SEQ ID NO: 23-25); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, a humanized immunoglobulin chain) containing CDR-L1, CDR-L2 and CDR-L3 of a VL domain containing SEQ ID NO: 27 (eg, SEQ ID NO: 28-30); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (for example, a humanized immunoglobulin chain) containing CDR-H1, CDR-H2 and CDR-H3 V _H -domain containing SEQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118 , 120 or 122 (for example, SEQ ID NO: 33, 34 (or 446, 454, 455, 456, 457 or 458) or 35); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The invention also relates to polypeptides (eg, a humanized immunoglobulin chain) containing CDR-L1, CDR-L2 and CDR-L3 of a VL domain containing SEQ ID NO: 37 or 126 (eg, SEQ ID NO: 38-40); or to any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 of these CDRs are variants of the sequence specified in the present description.

The present invention relates to crystalline compositions of antibodies against LAG3 and their antigennegative fragments of the present invention.

Polynucleotides.

In addition, the present invention provides polynucleotides encoding any of the anti-LAG3 antibody polypeptides or immunoglobulin chains and antigen-binding fragments thereof described herein (including the amino acid chain variants specifically mentioned herein). For example, the present invention relates to the polynucleotides described in

- 37 039293

SEQ ID NO: 1, 6, 11, 16, 21, 26

31, 36, 46, 48, 50, 52, 54, 56, 56, 58, 60, 62, 64, 66, 68, 70,

72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100,

102, 104, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123,125,

127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149.151,

153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175,177,

179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201,203,

205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227,229,

231, 233, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254,256,

258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280,282,

284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306,308,

310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332,334,

336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358,360,

362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384,386,

388, 390, 392, 394, 396, 398, 400, 402, or 404, and variants thereof (e.g., containing nucleotide sequences having at least 70%, 80%, 90%, 95%, or 99% BLAST sequence identity with such nucleotide sequences). sequences (as described above)); and polynucleotides encoding the amino acids described in the present description, for example, in

SEQID NO:

13, 33, 63, 95, 122

148

174

200

226

251

277

303

329

355

381

406

419

448

462

fourteen, :

34, ;

65,I

97,

124,

150,

176,

202,

228,

253,

279,

305,

331,

357,

383,

407,

426,

449,

463

fifteen,

35,

67,

17,

37,

69,

101

126

152

178

204

230

255

281

307

333

359

385

408

427

451 or

128

154

180

206

232

257

283

309

335

361

387

409

434

452

464 .

!, 19,

I, 39, ., 73, 103, , 130, 156, 182, 208, 234, 259, 285, 311, 337, 363, 389, 410, 435, 453

twenty,

40,

75, 106, 132, 158, 184, 210, 235, 261, 287, 313, 339, 365, 391, 411, 436, 454

22,

45,

77,

108:2

23,

47,

79,

134

160

186

212

237

263

289

315

341

367

393

412

437

455

3,

110

136

162

188

214

239

265

291

317

343

369

395

413

438

456

four, . 25, , 51, , 83,

112

138

164

190

216

241

267

293

319

345

371

397

414

439

457

5, 7

27,

53,

85,

114

140

166

192

218

243

269

295

321

347

373

399

415

440

458

eight,

28:

55, .

87,

116

142

168

194

220

245

271

297

323

349

375

401

416

441

459

9, , 10. , 30 , 59 , 91 118, 144, 170, 196, 222, 247, 273, 299, 325, 351, 377, 403, 417, 442, 460,

120

146

172

198

224

249

275

301

327

353

379

405

418

446

461

The scope of the present invention also includes polynucleotide variants that hybridize to any such polynucleotides.

Moreover, the present invention provides anti-LAG3 antibodies and antigen-binding fragments thereof comprising immunoglobulin heavy and light chains (eg, their variable regions) and/or heavy and light chain CDRs encoded by the polynucleotides described herein.

For example, the present invention relates to anti-LAG3 antibodies and antigen-binding fragments thereof, comprising an immunoglobulin heavy chain encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 107 (or encoding a variable domain thereof) and an immunoglobulin light chain encoded by the nucleotide sequence, specified in SEQ ID NO: 125 (or encoding its variable domain). For example, the present invention also provides anti-LAG3 antibodies and antigen-binding fragments thereof, comprising an immunoglobulin heavy chain encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 115 (or encoding its variable domain) and an immunoglobulin light chain encoded by the nucleotide sequence specified in SEQ ID NO: 125 (or encoding its variable domain).

The present invention relates to a polynucleotide encoding

- 38 039293

V _L 4A10 or its mature fragment;

VH 4A10 or a mature fragment thereof;

V _L 19E8 or its mature fragment;

V _H 19E8 or its mature fragment;

V _L 11C9 or its mature fragment;

V _H 11C9 or its mature fragment;

V _L -VH 22D2 or its mature fragment; where the V _L or VH domain of 4A10, 19E8, 11C9 and 22D2 is the V _L or VH domain of a mouse or a humanized V _L or VH domain of 4A10, 19E8, 11C9 and 22D2 described herein, and where optional V _L and/or VH is a variant of V _L or VH described in the present description.

The invention also relates to a polynucleotide containing the nucleotide sequence specified in SEQ ID NO: 1, or its nucleotides 58-408; or to its variant.

The invention also relates to a polynucleotide containing the nucleotide sequence specified in SEQ ID NO: 6, or its nucleotides 61-396; or to its variant.

The invention also relates to a polynucleotide containing the nucleotide sequence specified in SEQ ID NO: 11, or its nucleotides 58-414; or to its variant.

The invention also relates to polynucleotides containing the nucleotide sequence specified in SEQ ID NO: 16, or its nucleotides 61-384; or their variants.

The invention also relates to polynucleotides containing the nucleotide sequence specified in SEQ ID NO: 21, or its nucleotides 58-426; or their variants.

The invention also relates to polynucleotides containing the nucleotide sequence specified in SEQ ID NO: 26, or its nucleotides 61-384; or their variants.

The invention also relates to polynucleotides containing the nucleotide sequence specified in SEQ ID NO: 31, or its nucleotides 61-447; or their variants.

The invention also relates to polynucleotides containing the nucleotide sequence specified in SEQ ID NO: 36, or its nucleotides 61-547; or their variants.

The polynucleotide variants described herein include those that hybridize under low, moderate, or high stringency conditions to the polynucleotides described herein or to polynucleotides that encode the polypeptides described herein and encode anti-LAG3 antibody immunoglobulin chains or their antigen-binding fragments that retain the ability to specifically bind to LAG3 (human and/or cynomolgus macaque, eg Macaca fascicularis or Macaca mulatta). The first polynucleotide molecule is hybridizable to a second polynucleotide molecule when the single stranded form of the first polynucleotide molecule can be annealed to the second polynucleotide molecule under appropriate temperature and solution ionic strength conditions (see Sambrook, et al., supra). Temperature and ionic strength conditions determine the stringency of hybridization. Typical low stringency hybridization conditions include 55° C., 5X SSC, 0.1% SDS, and no formamide; or 30% formamide, 5X SSC, 0.5% SDS at 42°C. Typical hybridization conditions of moderate stringency are 40% formamide with 5X or 6X SSC and 0.1% SDS at 42°C. High stringency hybridization conditions are 50% formamide, 5X or 6X SSC at 42°C or optionally higher temperature (eg 57, 59, 60, 62, 63, 66 or 68°C). Typically, SSC is 0.15 M NaCl and 0.015 M Na-citrate. Hybridization requires that the two polynucleotides contain complementary sequences, although, depending on the stringency of the hybridization, mismatches between bases are possible. The appropriate stringency for nucleic acid hybridization depends on the length of the nucleic acids and the degree of complementarity, which are variables well known in the art. The higher the degree of similarity or homology between two nucleotide sequences, the higher is the stringency at which nucleic acids can hybridize. For hybrids longer than 100 nucleotides, equations have been derived to calculate the melting point (see Sambrook, et al., supra, 9.50-9.51). For hybridization with shorter polynucleotides, eg oligonucleotides, the position of the base mismatches becomes more important and the length of the oligonucleotide determines its specificity (see Sambrook, et al., supra, 11.7-11.8).

In another embodiment of the invention, a polynucleotide, eg DNA, encoding immunoglobulin polypeptide chains of anti-LAG3 antibodies or antigen-binding fragments described in the present invention is part of the present invention. In one embodiment, the polynucleotide encodes at least one mature immunoglobulin polypeptide light chain (VL) variable domain and at least one mature immunoglobulin polypeptide heavy chain variable domain (VH), wherein the VL domain comprises CDR-L1, CDR-L2, and CDR -L3 having a sequence selected from SEQ ID NOs: 8-10, 18-20, 28-30 and 38-40 and the VH domain contains CDRH1, CDR-H2 and CDR-H3 having a sequence selected from SEQ ID NO: 3-5, 13-15, 23-25 and 33-35. In one embodiment, the nucleic acid encodes the 4A10, 11C9, 19E8, or 22D2 light chain mature variable region and/or heavy variable region sequences.

- 39 039293 chain 4A10, 11C9, 19E8 or 22D2. In some embodiments, a polynucleotide encodes both a light chain and a heavy chain on the same polynucleotide molecule, and in other embodiments, the light and heavy chains are encoded on separate polynucleotide molecules, e.g., in separate or shared host cells. In another embodiment, the polynucleotides further encode a signal sequence.

In one embodiment, the polynucleotide encodes a mature immunoglobulin light chain (V _L ) variable domain comprising CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 7. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin light chain (V _L ) variable domain comprising CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 17. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin light chain (VL) variable domain comprising CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 27. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin light chain variable domain (V _L ) comprising CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 37 or 126. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin heavy chain (VH) variable domain comprising CDR-H1, CDR-H2, and CDR-H3 of SEQ ID NO: 2. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin heavy chain (VH) variable domain comprising CDR-H1, CDR-H2, and CDR-H3 of SEQ ID NO: 12. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes a mature immunoglobulin heavy chain (VH) variable domain comprising CDR-H1, CDR-H2, and CDR-H3 of SEQ ID NO: 22. Variants of such polynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a mature immunoglobulin heavy chain variable domain (V _H ) comprising CDR-H1, CDR-H2 and CDR-H3 SEQ ID NOs: 32, 106, 108, 110, 112, 114, 116, 118, 120 or 122. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes the light chain variable domain (V _L ) of an immunoglobulin of SEQ ID NOs: 7, 17, 27 and/or 37. Variants of such polynucleotides are also part of the present invention.

In one embodiment, the polynucleotide encodes an immunoglobulin heavy chain variable domain (V _H ) of SEQ ID NOs: 2, 12, 22, 32, 106, 108, 110, 112, 114, 116, 118, 120, and/or 122. Variants of such polynucleotides are also part of the present invention.

The present invention also relates to vectors, e.g. expression vectors such as plasmids, containing the polynucleotides of the invention (the sequences herein and variants thereof, e.g. SEQ ID NOs: 125, 105, 107, 109, 111, 113, 115 , 117, 119 and/or 121), wherein the polynucleotide is operably linked to control sequences that are recognized by the host cell when the host cell is transfected with the vector. Also provided are host cells containing a polynucleotide (for example, integrated into the genome, for example, a chromosome, of a host cell), or a vector of the present invention and methods for producing an antibody, or an antigen-binding fragment, or a polypeptide described in the present invention, including culturing a host cell containing an expressing a vector or polynucleotide encoding the immunoglobulin chains of the antibody or antigen-binding fragment thereof in the culture medium, and isolating the antigen or antigen-binding fragment thereof from the host cell or culture medium.

Binding affinity.

As a non-limiting example, anti-LAG3 antibodies and antigen-binding fragments thereof described herein bind human and/or cynomolgus monkey LAG3, e.g. Macaca fascicularis or Macaca mulatta LAG3 ^, e.g. ⁷ M); at least about 10 nM; or at least about 1 nM. In further embodiments, the antibodies have KD values of at least about 200 pM (2 x 10 ^-10 M), 100, 50, 20, 10, 5, or even 2 pM. For example, KD is approximately 2.77 x 10 ^-12 M, 1.47 x 10 ^-11 M, 1.47 x ^{10 -10} M, or 9.03 x 10 ^-11 M; or higher affinity. In one embodiment of the invention, KD is measured in a KinExA assay or similar kinetic exclusion assay. See, for example, Darling et al., Assay and Drug Dev. Tech. 2(6): 647-657 (2004).

Methods for producing antibodies and their antigen-binding fragments.

Hybridoma cells that produce parental (eg, mice) anti-LAG3 monoclonal antibodies, or an antigen-binding fragment thereof, as described herein, can be obtained by methods that are well known in the art. These isolated hybridomas are

- 40 039293 part of the present invention. These methods include, but are not limited to, the hybridoma method originally developed by Kohler, et al. (1975) (Nature 256:495-497), as well as the triome method (Hering, et al. (1988) Biomed. Biochim. Acta. 47:211-216 and Hagiwara, et al. (1993) Hum. Antibod. Hybridomas 4:15), human B-cell hybridoma method (Kozbor, et al. (1983) Immunology Today 4:72 and Cote, et al. (1983) Proc. Natl. Acad. Sci. U.S.A 80:2026-2030), EBV hybridoma method (Cole, et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., p. 77-96, 1985) and electric field-based electrofusion using the Cyto Pulse (Cyto Pulse Sciences, Inc., Glen Burnie, MD). Preferably, mouse splenocytes are isolated and fused by PEG or by electrofusion with mouse myeloma cells, based on standard protocols. The resulting hybridomas can then be screened for the production of antigen-specific antibodies. For example, suspensions of single spleen lymphocytes from immunized mice can be fused with one sixth of non-secreting P3X63Ag8.653 mouse myeloma cells (ATCC, CRL 1580) using 50% PEG. Cells can be seeded at approximately 2x105 cells/mL in a flat bottom microtiter plate followed by incubation for two weeks in selective medium containing 20% fetal clonal serum, 18% 653 conditioned medium, 5% origen (IGEN), 4 mM L-glutamine, 1 mM L-glutamine, 1 mM sodium pyruvate, 5 mM HEPES, 0.055 mM 2-mercaptoethanol, 50 U/mL penicillin, 50 mg/mL streptomycin, 50 mg/mL gentamicin, and 1X HAT (Sigma; HAT added 24 hours after confluence). After two weeks, cells can be cultured in medium in which HAT has been replaced with HT. Individual wells can then be screened by ELISA for anti-LAG3 IgG monoclonal antibodies. After extensive growth of hybridomas occurs, the environment can usually be observed after 10-14 days. Antibody-secreting hybridomas can be replated, rescreened and, if still positive for anti-LAG3 human IgG monoclonal antibodies, can be subcloned at least twice by the limiting dilution method. The stable subclones can then be cultured in vitro to generate small amounts of antibody in tissue culture medium for characterization.

Thus, the present invention relates to methods for producing an anti-LAG3 antibody or antigen-binding fragment of the present invention, comprising culturing a hybridoma cell that expresses the antibody or fragment under conditions conducive to such expression, and optionally isolating the antibody or fragment from the hybridoma.

The anti-LAG3 antibodies described herein can also be produced by recombinant methods (eg, in an E. coli/T7 expression system). In this embodiment, nucleic acids encoding anti-LAG3 antibody molecules of the invention (e.g., V _H or V _L ; e.g., any one or more of SEQ ID NOs: 125, 105, 107, 109, 111, 113, 115, 117, 119 and/or 121) can be inserted into a pET-based plasmid and expressed in the E. coli/T7 system. For example, the present invention relates to methods for expressing an antibody, or an antigen-binding fragment thereof, or an immunoglobulin chain thereof, in a host cell (e.g., a bacterial host cell such as E. coli, such as BL21 or BL21DE3), comprising the expression of an RNA polymerase T7 in a cell that also includes a polynucleotide encoding an immunoglobulin chain that is operably linked to the T7 promoter. For example, in one embodiment, a bacterial host cell, such as E. coli, comprises a polynucleotide encoding a T7 RNA polymerase gene operably linked to a lac promoter, and polymerase and strand expression is induced by incubating the host cell with IPTG .

Several methods for producing recombinant antibodies are known in the art. One example of a method for recombinant antibody production is described in US Pat. No. 4,816,567.

Transformation can be carried out by any known method of introducing polynucleotides into the host cell. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of polynucleotide(s) into liposomes, biolistic injection, and direct microinjection of DNA into nuclei. In addition, nucleic acid molecules can be introduced into mammalian cells via viral vectors. Methods for transforming cells are well known in the art. See, for example, US Pat. Nos. 4,399,216; 4912040; 4740461 and 4959455.

Thus, the present invention relates to recombinant methods for producing an anti-LAG3 antibody or an antigen-binding fragment of the present invention, or an immunoglobulin chain thereof, comprising (i) introducing a polynucleotide (e.g., any one or more of SEQ ID NOs: 125, 105, 107, 109, 111, 113, 115, 117, 119, and/or 121) encoding one or more immunoglobulin chains of an antibody or fragment (e.g., immunoglobulin heavy chain 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9 and/or immunoglobulin light chain 4A10, 19E8, 11C9 and/or 22D2; e.g. Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9 ), for example, where the polynucleotide is in the vector and/or is operably linked to a promoter; (ii) culturing the host cell (eg CHO, or Pichia or Pichia pastoris) under conditions

- 41 039293 facilitating the expression of the polynucleotide and, (iii) optionally, isolating the antibody or fragment or chain from the host cell and/or the medium in which the host cell was grown. Upon receipt of an antibody or antigen-binding fragment containing more than one immunoglobulin chain, for example, an antibody that contains two immunoglobulin heavy chains and two immunoglobulin light chains, co-expression of the chains in the same host cell leads to association of the chains, for example, in the cell, or on the surface cell, or outside the cell if such chains are secreted, thus forming an antibody molecule or antigen-binding fragment. Methods include methods in which only the immunoglobulin heavy chain or only the immunoglobulin light chain (eg, any of the chains described in the present invention, including its mature fragments and/or variable domains) is expressed. Such chains are useful, for example, as intermediates in the expression of an antibody or antigen-binding fragment that includes such a chain. For example, the present invention also provides anti-LAG3 antibodies and antigen-binding fragments thereof, comprising an immunoglobulin heavy chain (or variable domain or CDR thereof) encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 115 (or the variable domain encoding them) and an immunoglobulin light chain (or variable domain or CDR thereof) encoded by the nucleotide sequence set forth in SEQ ID NO: 125 (or the variable domain encoding them) that are the product of such production methods, and optionally purification methods, described in the present invention. .

Anti-LAG3 antibodies can also be synthesized by any of the methods described in US Pat. No. 6,331,415.

Eukaryotic and prokaryotic host cells, including mammalian cells, as hosts for expressing anti-LAG3 antibodies or immunoglobulin fragments or chains described in the present invention are well known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC). These include, among others, Chinese Hamster Ovary (CHO) cells, NSO, SP2 cells, HeLa cells, Baby Hamster Kidney (BHK) cells, Monkey Kidney (COS) cells, human hepatocellular carcinoma cells (e.g. Hep G2), A549 cells, 3T3 cells, HEK-293 cells and a number of other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, porcine, goat, bovine, horse and hamster cells. Particularly preferred cell lines are selected by determining which cell lines have high levels of expression. Other cell lines that can be used are insect cell lines (eg Spodoptera frugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plant cells and fungal cells. Fungal cells include yeast and filamentous fungi, including, for example,

Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri) , Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp. , Kluyveromyces lactis, Candida albicans , Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp.. Fusarium gramineum, Fusarium venenatum, Physcomitrella patens and Neurospora crassa. Pichia sp. , any Saccharomyces sp. , Hansenula polymorpha, any Kluyveromyces sp., Candida albicans , any Aspergillus sp., Trichoderma reesei, Chrysosporium lucknowense, any Fusarium sp. , Yarrowia lipolytica and Neurospora crassa.

In addition, the expression of antibodies and their antigen-binding fragments, and chains of immunoglobulins according to the invention (or other parts thereof) from producing cell lines can be increased using a number of known methods. For example, the glutamine synthetase gene expression system (GS system) is a common approach to enhance expression under certain conditions. The GS system is discussed in whole or in part in European Patent Nos. 0216846, 0256055 and 0323997 and European Patent Application No. 89303964.4. Thus, in one embodiment of the invention, mammalian host cells (e.g., CHO) lack the glutamine synthetase gene and are grown in the absence of glutamine in a medium where, however, the polynucleotide-encoding immunoglobulin chain contains the glutamine synthetase gene, which complements the absence of this gene in the host cell.

The present invention relates to methods for purifying an anti-LAG3 antibody or antigen-binding fragment thereof according to the present invention, including the introduction of a sample (for example, a cult

- 42 039293 cell lysate or cell lysate fraction, e.g. soluble lysate fraction) containing the antibody or fragment) into a purification medium (e.g. cation exchange medium, anion exchange medium, hydrophobic exchange medium, affinity purification medium (e.g. protein A , protein G, protein A/G, protein L)), and either collection of purified antibody or fragment from the past fraction of the specified sample, which is not associated with the medium; or discarding the flow-through fraction and eluting the bound antibody or fragment from the medium and collecting the eluate. In one embodiment of the invention, the medium is in the column into which the sample is placed. In one embodiment of the invention, the purification method is performed after recombinant expression of the antibody or fragment in a host cell, for example, where the host cell is first lysed and optionally the lysate is purified of insoluble materials prior to media purification; or where the antibody or fragment is secreted into the culture medium by the host cell and the medium or fraction thereof is applied to the medium for purification.

Typically, glycoproteins produced in a particular cell line or transgenic animal have a glycosylation pattern that is characteristic of glycoproteins produced in the cell line or transgenic animal. Thus, the specific glycosylation pattern of an antibody will depend on the particular cell line or transgenic animal used to produce the antibody. However, all antibodies encoded by the nucleic acid molecules described in the present invention, or containing the amino acid sequences described in the present description, are included in the present invention regardless of the nature of glycosylation, which may have antibodies. Similarly, in specific embodiments, antibodies with a glycosylation pattern that includes only non-fucosylated N-glycans may be advantageous as these antibodies have been shown to generally exhibit superior efficacy than their fucosylated counterparts, both in vitro and in vitro. vivo (See, for example, Shinkawa et al., J. Biol. Chem. 278: 3466-3473 (2003); US Pat. Nos. 6,946,292 and 7,214,775). These non-fucosylated N-glycan antibodies are unlikely to be immunogenic because their carbohydrate structures are a normal component of the IgG population that exists in human serum.

The present invention relates to antibodies against LAG3 and their antigen-binding fragments (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) containing N- linked glycans that are typically added to immunoglobulins produced in Chinese hamster ovary cells (CHO N-linked glycans) or engineered yeast cells (engineered yeast N-linked glycans), such as, for example, Pichia pastoris. For example, in one embodiment, the antibody or antigen binding fragment comprises one or more of the yeast engineered N-linked glycans or CHO N-linked glycans as indicated in FIG. 6 (eg GO and/or G0-F and/or G1 and/or G1-F and/or G2-F and/or Man5). In one embodiment, the antibody or antigen-binding fragment contains N-linked glycans modified by yeast engineering techniques, i. G0 and/or G1 and/or G2, optionally further including Man5. In one embodiment, the antibody or antigen-binding fragment contains N-linked CHO glycans, ie. G0-F, G1-F and G2-F, optionally further including G0 and/or G1 and/or G2 and/or Man5. In one embodiment, from about 80% to about 95% (e.g., about 80-90%, about 85%, about 90%, or about 95%) of all N-linked glycans on an antibody or antigen-binding fragment or immunoglobulin chains, are engineered yeast N-linked glycans or CHO N-linked glycans. See Nett et al., Yeast. 28(3): 237-252 (2011); Hamilton et al., Science. 313(5792): 1441-1443 (2006); Hamilton et al., Curr Opin Biotechnol. 18(5): 387-392 (2007). For example, in one embodiment, the engineered yeast cell is GFI5.0, or YGLY8316, or the lines of US Pat. No. 7,795,002 or Zha et al., Methods Mol Biol. 988:31-43 (2013). See also International Patent Application Publication No. WO 2013/066765.

The present invention relates to polyclonal anti-LAG3 antibodies and antigen-binding fragments thereof, e.g., a composition comprising a plurality of anti-LAG3 antibodies and fragments that include one or more of the anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9), and methods of using them. A polyclonal antibody is an antibody that is produced among or in the presence of one or more other non-identical antibodies. Typically, polyclonal antibodies are produced from collections of different B lymphocytes, eg, B lymphocytes from an animal that is injected with an immunogen of interest that produces a population of different antibodies, all of which are directed to the immunogen. Usually polyclonal antibodies are obtained directly from the immunized animal, for example, from the spleen, serum or ascitic fluid.

The present invention relates to anti-LAG3 antagonist antibodies and antigen-binding fragments thereof, and methods of using them, for example, to humanized anti-LAG3 antagonist antibodies and fragments. Antagonist antibody against LAG3 or antigen-bindings thereof

- 43 039293 the generating fragment antagonizes the activity of LAG3 (for example, human LAG3), for example, by inhibiting the binding of LAG3 to class II MHC molecules; by competition with MHC class II molecules for LAG3 binding; or, when the cell or individual is brought into contact with the antibody or fragment, a biological phenotype associated with LAG3 antagonism is obtained, such as stimulation of IL-2 production by antigen-specific T cells.

The present invention relates to bispecific and bifunctional antibodies and antigen-binding fragments having binding specificity for LAG3 and another antigen, such as PD-1 or PD-L1, for example, and methods of using them. In one embodiment, the anti-PD1 chains comprise the amino acid sequence of SEQ ID NOS: 41 and 42 or SEQ ID NOS: 43 and 44. A bispecific or bifunctional antibody is an artificial hybrid antibody having two different heavy/light chain pairs and two different binding sites . Bispecific antibodies can be generated in a variety of ways, including hybridoma fusion and Fab' binding. See, for example, Songsivilai, et al. (1990) Clin. Exp. Immunol. 79: 315-321, Kostelny, et al. (1992) J Immunol. 148:1547-1553. In addition, bispecific antibodies can be formed as diabodies (Holliger, et al. (1993) PNAS USA 90:6444-6448) or as Janusin (Traunecker, et al. (1991) EMBO J. 10:3655-3659 and Traunecker, et al (1992) Int J Cancer Suppl 7:51-52).

In addition, the present invention relates to antigennegative fragments against LAG3 antibodies against LAG3 described in the present invention. Antibody fragments include F(ab) ₂ - fragments that can be obtained by enzymatic cleavage of IgG, for example, pepsin. Fab fragments can be obtained, for example, by reduction of F(ab) ₂ dithiothreitol or mercaptoethylamine. The Fab fragment is a VL-CL chain attached to the V _H -C _H1 chain by a disulfide bond. The F(ab) ₂ fragment is two Fab fragments, which in turn are linked by two disulfide bridges. The Fab part of the F(ab)2 molecule includes part of the Fc region, between which there are disulfide bridges. An Fv fragment is a VL or VH region.

Immunoglobulins can be assigned to different classes depending on the amino acid sequences of the constant domain of their heavy chains. There are at least five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and some of these can be further subdivided into subclasses (isotypes), such as IgG-1, IgG-2, IgG-3, and IgG-4 ; IgA-1 and IgA-2. The invention relates to antibodies against LAG3 and antigen-binding fragments of any of these classes or subclasses of antibodies.

In one embodiment, the anti-LAG3 antibody or antigen-binding fragment comprises a heavy chain constant region, eg, a human constant region, such as a human γ1, γ2, γ3, or γ4 heavy chain constant region or a variant thereof. In another embodiment, the anti-LAG3 antibody or antigen-binding fragment comprises a light chain constant region, eg, a human light chain constant region, such as a human lambda or kappa light chain region. As a non-limiting example, the human heavy chain constant region may be γ4 and the human light chain constant region may be kappa. In an alternative embodiment, the Fc region of the antibody is γ4 with a Ser228Pro mutation (Schuurman, J et. al., Mol. Immunol. 38: 1-8, 2001).

In some embodiments, different constant domains can be attached to humanized V _L and V _H regions derived from the CDRs described in the present invention. For example, if the specific intended use of an antibody (or fragment) of the present invention is to provide altered effector functions, a heavy chain constant domain other than human IgG1 may be used, or an IgG1/IgG4 hybrid may be used.

Although human IgG1 antibodies provide a long half-life and effector functions such as complement activation and antibody-dependent cellular cytotoxicity, such activity may not be desirable for all uses of the antibody. In such cases, for example, a human IgG4 constant domain can be used. The present invention relates to anti-LAG3 antibodies and antigen-binding fragments thereof, which contain an IgG4 constant domain, eg, humanized anti-LAG3 antagonist antibodies and fragments, and methods of using the same. In one embodiment, the IgG4 constant domain may differ from the native human IgG4 constant domain (Swiss-Prot accession number P01861.1) at a position corresponding to position 228 in the EU system and position 241 in the CAVAT system, where native Ser108 is replaced by Pro, for preventing the potential formation of an interchain disulfide bond between Cys106 and Cys109 (corresponding to the positions of Cys226 and Cys229 in the EU system and the positions of Cys239 and Cys242 in the CABAT system), which may prevent the formation of proper intrachain disulfide bonds. See Angal et al. (1993) Mol. Imunol. 30:105. In other cases, a modified IgG1 constant domain that has been modified to increase half-life or decrease effector function can be used.

Antibody engineering.

Also included are embodiments in which anti-LAG3 antibodies and their antigen binding

- 44 039293 calling fragments are engineered antibodies that include modifications to framework residues in the variable domains of the parent (eg, mouse) monoclonal antibody, for example, to improve the properties of the antibody or fragment. Typically, such framework modifications are made to reduce the immunogenicity of the antibody or fragment. This is typically done by replacing non-CDR residues in the variable domains (i.e., framework residues) in the parent (e.g., rodent) antibody or fragment with similar residues from the immune repertoire of the species in which the antibody is intended to be used, e.g., human residues in the case of medicines for humans. Such an antibody or fragment is referred to as a humanized antibody or fragment. In some instances, it is desirable to increase the affinity or change the specificity of an engineered (eg, humanized) antibody. One approach is to back-mutate one or more framework residues to the appropriate germline sequence. More specifically, an antibody or fragment that has undergone a somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the framework sequences of the antibody or fragment with the germline sequences from which the antibody or fragment is derived. Another approach is to return to the original parental (eg, rodent) residue at one or more positions of the engineered (eg, humanized) antibody, eg, to restore binding affinity that may be lost during the replacement of framework residues. (See, for example, US patent No. 5693762, US patent No. 5585089 and US patent No. 5530101).

For example, in table. 2 below shows the regions where the amino acid position in the framework region (using the Kabat numbering system) differs from the germline, and how that position can be backmutated to the germline by these substitutions:

table 2

Illustrative backmutations

Region Amino acid position of the scaffold regions (Kabat numbering) Back Mutation Examples V _H ALA 25 H25S V _H ALA 68 S68T V _H ALA 82a T82aT

Another type of framework modification involves mutating one or more residues in the framework or even one or more CDR regions to remove T-cell epitopes, thereby reducing the potential immunogenicity of the antibody. This approach is also referred to as deimmunization and is described in more detail in US Pat. No. 7,125,689.

In specific embodiments, it is desirable to change certain amino acids containing exposed side chains to a different residue in order to provide a higher chemical stability of the final antibody, as follows. Such changes in the antigen-binding region may alter antigen binding. Deamidation of asparagine can occur in the N-G or D-G sequences and can lead to the formation of an isoaspartic acid residue that introduces a loop into the polypeptide chain and reduces its stability (the isoaspartic acid effect). In certain embodiments, the antibodies of the present invention do not contain asparagine isomerism regions.

For example, an asparagine (Asn) residue can be replaced with Gln or Ala to reduce the potential for isoaspartate formation in any Asn-Gly sequences, in particular in CDRs. A similar problem can occur in the Asp-Gly sequence. Reissner and Aswad (2003) Cell. Mol. life sci. 60:1281.

The formation of isoaspartate can deplete or completely eliminate the binding of an antibody to its target antigen. See, Presta (2005) J. Allergy Clin. Immunol. 116:731, 734. In one embodiment, asparagine is replaced by glutamine (Gln). It may also be desirable to change the amino acid adjacent to an asparagine (Asn) or glutamine (Gln) residue to reduce the likelihood of deamidation, which occurs more rapidly when smaller amino acids are adjacent to asparagine or glutamine. See, Bischoff & Kolbe (1994) J. Chromatog. 662:261. In addition, any methionine residues (typically solvent-exposed to Met) in the CDR can be replaced with Lys, Leu, Ala, or Phe to reduce the chance that the methionine sulfur will be oxidized, which can reduce antigen binding affinity and also contribute to molecular heterogeneity in the final production of antibodies. There. In one embodiment, methionine is replaced by alanine (Ala). In addition, to prevent or minimize potential cleavage of Asn-Pro peptide bonds, it may be desirable to change any combinations of Asn-Pro found in CDRs to Gln-Pro, Ala-Pro, or Asn-Ala. Antibodies with these substitutions are then screened to ensure that the substitutions do not reduce the affinity or specificity of the antibody for LAG3,

- 45 039293 or other desired biological activity to unacceptable levels.

Table 3

Exemplary Stabilizing CDR Variants

stabilizing

The rest of the CDR sequence

Asn-Gly

Gln-Gly, Ala-Gly or Asn-Ala

Asp-Gly

Glu-Gly, Ala-Gly or Asp-Ala

Lys, Leu, Ala or Phe exposed solvent;

Gin or Ala

O or A

The 4A10, 19E8, 11C9 and 22D2 immunoglobulin chains mentioned above contain residues that are n-Pro, Ala-Pro or Asn-Ala

QG,

A-G or N-A

Q-P ,

A-P or N-A

K , L ,

A or F

E-G,

A-G or D-A are underlined with a double line. The scope of the present invention includes antibodies, antigen-binding fragments, polypeptides, and polynucleotides as described herein, wherein any one or more of such residues are mutated to any other residue, including, for example, the stabilizing variant sequence residues identified in Table 1 above. 3.

Anti-mouse LAG3 antibodies and antigen-binding fragments can be humanized by various methods known in the art (see, for example, humanization methods described in WO 2005/047326 or US7846443). For example, in one embodiment, mouse anti-LAG3 antibodies and fragments are humanized by a method that uses computer molecular modeling to identify CDR loops in non-human immunoglobulin chains. This identification is based on the three-dimensional structure of the immunoglobulin chain and the position of the loops in the chain.

Human framework regions (obtained from the IMGD database) into which non-human loops are inserted are selected based on the best matches (by amino acid sequence comparison) with the non-human sequence in both framework regions and CDRs. With respect to FR4 in the VH domain, the VJ regions for human germline sequences are compared to the corresponding non-human VJ regions; and, in the case of FR4 in the VL domain, the J-kappa and J-lambda regions of the human germline sequences are compared to the corresponding non-human J-kappa and J-lambda regions.

Proper three-dimensional orientation of the CDR, which is critical for maintaining antigen binding, depends in part on a proper contact surface between VH and VL. Thus, molecular modules are designed and used to identify residues at the VL-VH interface, as well as to identify residues that can potentially change CDR conformation and thus antigen binding. If necessary, mutations in the immunoglobulin chain can be introduced so as to achieve the desired properties, such as antigen binding.

Using molecular modeling, design capability filters are established. Design capability filters are criteria used to eliminate features from the final immunoglobulin chain in order to avoid unwanted effects. Molecular models are also used to identify solvent-exposed amino acids that can lead to undesirable effects such as glycosylation, deamidation and oxidation. Such effects on an antibody can lead to a change in the conformation of the antibody and thus its function. Such problems can occur, for example, during scaling up or over a long period of time under the influence of excessive chemical / physical conditions of the environment. Again, if necessary, mutations in the chains can be made to achieve the desired properties.

Design capability filters are typically used early in the humanized circuit modeling stage to eliminate/minimize these potential problems.

Humanized antibodies are further evaluated using modeling criteria such as high expressibility and desired isoelectric points.

Engineering Fc-regions of antibodies.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can also

- 46 039293 but designed to include modifications to the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or effector function (e.g., antigen-dependent cellular cytotoxicity ). Moreover, antibodies and their antigen-binding fragments (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, and/or Ab9) can be chemically modified (e.g., one or more chemical moieties may be linked to an antibody) or may be modified to alter their glycosylation, again to alter one or more functional properties of the antibody or fragment. Each of these embodiments is described in more detail below. The numbering of the residues in the Fc regions corresponds to the EU index according to Kabat. Any of such anti-LAG3 antibodies, or antigen-binding fragments thereof, having the modifications (eg, Fc modifications) and/or changes described herein is part of the present invention.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) also include antibodies and fragments with modified (or blocked) Fc regions to provide altered effector functions. See, for example, US patent No. 5624821; WO2003/086310; WO2005/120571; W02006/0057702. Such modifications can be used to enhance or suppress various responses of the immune system, with possible beneficial effects in diagnosis and therapy. Changes in the Fc region include amino acid changes (substitutions, deletions and insertions), glycosylation or deglycosylation, and the addition of several Fcs. Fc changes can also change the half-life of antibodies in therapeutic antibodies, allowing for less frequent dosing and thus increased convenience and reduced material usage. See Presta (2005) J. Allergy Clin. Immunol. 116:731, 734-35.

In one embodiment, an anti-LAG3 antibody (e.g., humanized antibodies, such as humanized antagonist antibodies) or an antigen binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is an IgG4 isotype antibody or fragment containing a serine to proline mutation at a position corresponding to position 228 (S228P; index EU) in the hinge region of the heavy chain constant region. This mutation reportedly eliminates the heterogeneity of disulfide bridges between heavy chains in the hinge region (Angal et al. supra; position 241 is based on the Kabat numbering system).

In one embodiment of the invention, the CH1 hinge region is modified such that the number of cysteine residues in the hinge region is increased or decreased. This approach is further described in US Pat. No. 5,677,425. The number of cysteine residues in the CH1 hinge region is changed, for example, to facilitate assembly of light and heavy chains, or to increase or decrease antibody stability.

In another embodiment, the Fc hinge region of an anti-LAG3 antibody or antigen-binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, and/or Ab9) is introduced a mutation to reduce the biological half-life of the antibody or fragment. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface of the Fc-hinge fragment such that the antibody or fragment has reduced staphylococcal protein A (SpA) binding relative to that of the native Fc-hinge domain to SpA. This approach is described in more detail in US Pat. No. 6,165,745.

In another embodiment, an anti-LAG3 antibody (e.g., humanized antibodies, such as humanized antagonist antibodies) or an antigen-binding fragment thereof (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8 and/or Ab9) are modified to increase their biological half-life. Various approaches are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in US Pat. No. 6,277,375. from two loops of the CH2 domain of the IgG Fc region, as described in US patents No. 5869046 and 6121022.

In other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector function(s) of the anti-LAG3 antibody or antigen-binding fragment. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, and 322 can be replaced with a different amino acid residue such that the antibody has an altered affinity for the effector ligand but retains the ability to bind the parent antigen. antibodies. The effector ligand for which the affinity is being changed may be, for example, the Fc receptor or the C1 component of complement. This approach is described in more detail in US Pat. Nos. 5,624,821 and 5,648,260.

- 47 039293

In another example, one or more amino acids selected from amino acid residues 329, 331, and 322 can be replaced with a different amino acid residue such that the anti-LAG3 antibody has altered C1q binding and/or reduced or eliminated complement dependent cytotoxicity (CDC). This approach is described in more detail in US Pat. No. 6,194,551.

In one example, one or more amino acid residues at amino acid positions 231 and 239 are altered, thereby altering the ability of the anti-LAG3 antibody or antigen-binding fragment thereof to fix complement. This approach is further described in PCT publication WO 94/29351.

In another example, the Fc region is modified to reduce the ability of an anti-LAG3 antibody (e.g., humanized antibodies, such as humanized antagonist antibodies) or an antigen-binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) mediate antibody-dependent cellular cytotoxicity (ADCC) and/or reduce the affinity of an antibody or fragment for the Fcy receptor by modifying one or more amino acids at the following positions:

238, 239, 243,

248, 249, 252, 254, 255, 256, 258, 264, 265, 267, 268, 269.270,

272, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293, 294,295,

296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322, 324,326,

327, 329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373,376,

378, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439.

This approach is further described in PCT publication WO 00/42072.

Moreover, binding sites on human IgG1 for FcyRI, FcyRII, FcyRIII and FcRn have been mapped and variants with improved binding have been described (see Shields et al. (2001) J. Biol. Chem. 276:6591-6604).

In one embodiment, the Fc region is modified to reduce the ability of an anti-LAG3 antibody (e.g., humanized antibodies, such as humanized antagonist antibodies) or antigen-binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) mediate effector function and/or enhance anti-inflammatory properties by modifying residues 243 and 264. In one embodiment, the Fc region of an antibody or fragment is modified by changing residues at positions 243 and 264 for alanine. In one embodiment, the Fc region is modified to reduce the ability of the antibody or fragment to mediate effector function and/or enhance anti-inflammatory properties by modifying residues 243, 264, 267, and 328.

In another embodiment, an anti-LAG3 antibody (e.g., humanized antibodies such as humanized antagonist antibodies) or an antigen binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) have specific glycosylation patterns. For example, an aglycosylated antibody or fragment (i.e., the antibody is devoid of glycosylation) can be prepared. The glycosylation pattern of an antibody or fragment can be altered, for example, to increase the affinity or avidity of the antibody or fragment for the LAG3 antigen. Such modifications can be made, for example, by changing one or more glycosylation sites in the sequence of the antibody or fragment. For example, one or more amino acid substitutions can be made that result in the elimination of one or more glycosylation sites in the framework region, thereby eliminating glycosylation in that region. Such aglycosylation may increase the affinity or avidity of an antibody or fragment for an antigen. See, for example, US Pat. Nos. 5,714,350 and 6,350,861.

Anti-LAG3 antibodies (for example, humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments described herein (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8 and/or Ab9) include antibodies and fragments produced in lower eukaryotic host cells, in particular fungal host cells such as yeast (e.g. Pichia pastoris) and filamentous fungi that are genetically modified to produce glycoproteins that have a glycosylation pattern similar to that of mammals or humans (See, for example, Choi et al. (2003) Proc. Natl. Acad. Sci. 100: 5022-5027; Hamilton et al. (2003) Science 301: 1244-1246; Hamilton et al (2006) Science 313: 1441-1443). A particular disadvantage of these genetically modified host cells over currently used mammalian cell lines is the ability to control the glycosylation profile of the glycoproteins that are produced in the cells, so that glycoprotein compositions can be prepared in which a particular N-glycan structure predominates (see, for example, patent US No. 7029872 and US patent # 7449308). These genetically modified host cells are used to generate antibodies that generally have specific N-glycan structures (see, for example, Li et al. (2006) Nat. Biotechnol. 24: 210-215).

- 48 039293

In specific embodiments, anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) further includes antibodies and fragments produced in lower eukaryotic host cells and which contain fucosylated and non-fuosylated hybrid and complex N-glycans, including bisector and multiantennary types, including but not limited to without limitation, N-glycans such as

GlcNAc (i-4)Man ₃ GlcNAc2 7 Ga 1 (1-4; GlcNAc (i-4)Man ₃ GlcNAc ₂ ! NANA(1-4 j Gal (1-4; GlcNAc (i-4)Man ₃ GlcNAc ₂ .

In specific embodiments, anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may contain antibodies or fragments having at least one hybrid N-glycan selected from the group consisting of

GlcNAcMan ₅ GlcNAc ₂ ; GalGlcNAcMan ₅ GlcNAc ₂ and NANAGalGlcNAcMan ₅ GlcNAc ₂ .

In specific aspects, the hybrid N-glycan is the predominant type of N-glycan in the composition. In the following aspects, a hybrid N-glycan is a particular type of N-glycan that is about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% or 100% of the hybrid N-glycans in the composition.

In specific embodiments, anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) contain antibodies and fragments having at least one N-glycan complex selected from the group consisting of

GlcNAcMan ₃ GlcNAc ₂ ; GalGlcNAcMan ₃ GlcNAc ₂ ; NANAGalGlcNAcMan ₃ GlcNAc ₂ ;

GlcNAc ₂ Man ₃ GlcNAc ₂ ; GalGlcNAc ₂ Man ₃ GlcNAc ₂ ; Gal ₂ GlcNAc2Man ₃ GlcNAc2 ;

NANAGal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ and NANA ₂ Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ .

In specific aspects, the complex N-glycan is the predominant type of N-glycans in the composition. In the following aspects, the complex N-glycan is a specific type of N-glycan that is 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% or 100% of the complex N-glycans in the composition.

In specific embodiments, the N-glycan of the anti-LAG3 antibody and antigen-binding fragment is fucosylated. Typically, fucose has an a1,3 bond to GlcNAc at the reducing end of the N-glycan, an a1,6 bond to GlcNAc at the reducing end of the N-glycan, an a1,2 bond to Gal at the non-reducing end of the N-glycan, a1,3 -bond to GlcNac at the non-reducing end of the N-glycan or a1,4-bond to GlcNAc at the non-reducing end of the N-glycan.

Thus, in specific aspects of the glycoprotein compositions described above, the glycoform has a fucose in the α1,3 bond or α1,6 bond to form a glycoform selected from the group consisting of

Man ₅ GlcNAc ₂ (Fuc) , GlcNAcMan ₅ GlcNAc ₂ (Fuc) ,

Man ₃ GlcNAc ₂ (Fuc) , GlcNAcMan ₃ GlcNAc ₂ (Fuc) , GlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) ,

GalGlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) , Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) ,

NANAGal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) and NANA ₂ Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) ; NANA ₂ Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ (Fuc) ;

fucose in an α1,3 bond or a1,4 bond to form a glycoform selected from the group consisting of GlcNAc (Fuc) Man ₅ GlcNAc ₂ , GlcNAc (Fuc) Man ₃ GlcNAc ₂ , GlcNAc ₂ (Fuci- ₂ ) Man _{3 GlcNAc2} ,

Ga 1G1 cNAc ₂ ( Fuc 1-2 ) Man ₃ GlCNAC2 r Ga I2GICNAC2 (Fuc 2 ) Man3G1 cNAc2 _f

NANAGal2GlcNAc2 (Fuci- ₂ ) Man ₃ GlcNAc ₂ and NANA ₂ Gal ₂ GlcNAc ₂ (Fuci_ ₂ ) Man ₃ GlcNAc ₂ ;

or fucose in an α 1,2 bond to form a glycoform preparation selected from the group consisting of Gal (Fuc) GlcNAc ₂ Man ₃ GlcNAc ₂ , Gal ₂ (Fuci- ₂ ) GlcNAc ₂ Man ₃ GlcNAc ₂ ,

NANAGal ₂ (FuCi-2) GlcNAc ₂ Man ₃ GlcNAc ₂ and NANA ₂ Gal ₂ (FuCi- ₂ ) GlcNAc ₂ Man ₃ GlcNAc ₂ .

In the following aspects, anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antagonist antibodies) or antigen-binding fragments thereof contain high mannose N-glycans, including, but not limited to, Man ₈ GlcNAc2, Man ₇ GlcNAc2, Man ₆ GlcNAc2 , Man ₅ GlcNAc ₂ , Man ₄ GlcNAc ₂ , or N-glycans, which consist of the Man ₃ GlcNAc2 N-glycan structure. In further aspects of the above, complex N-glycans further include fucosylated and non-fucosylated bisector and multiantennary types.

In one embodiment, the LAG3 antibody or antigen-binding fragment thereof (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, or Ab9) contains an immunoglobulin Fc domain that contains glycans that contain sialic acid (e.g., Nacetylneuraminic acid), for example, terminal a2,3-sialic acid or terminal α2,6

- 49 039293 sialic acid. In one embodiment, the glycans on Fc are types sialylated at 5%, 10%, 20%, 50%, 90% or more. In one embodiment, the Fc contains mutations at positions 297, 264 and/or 243.

As used herein, the terms N-glycan and glycoform are used interchangeably and refer to an N-linked oligosaccharide, for example an oligosaccharide that is linked by an asparagine-N-acetylglucosamine bond to an asparagine residue on a polypeptide. N-linked glycoproteins contain an N-acetylglucosamine residue linked to the amide nitrogen of an asparagine residue in the protein. The predominant sugars found on glycoproteins are glucose, galactose, mannose, fucose, N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc) and sialic acid (eg N-acetyl-neuraminic acid (NANA)). Processing of sugar groups for N-linked glycoproteins occurs during translation in the ER lumen and continues post-translationally in the Golgi apparatus.

N-glycans share a pentasaccharide core from Man ₃ GlcNAc2 (Man refers to mannose; Glc refers to glucose; and NAc refers to N-acetyl; GlcNAc refers to Nacetylglucosamine). Typically, N-glycan structures have a non-reducing end on the left and a reducing end on the right. The reducing end of an N-glycan is the end that is linked to an Asn residue containing a glycosylation site on a protein. N-glycans differ in the number of branches (antennae) containing peripheral sugars (e.g., GlcNAc, galactose, fucose, and sialic acid) that are added to the structure of the Man ₃ GlcNAc2 (Man3) core, which is also called the trimannose core, pentasaccharide core, or oligomannose core. N-glycans are classified according to their branched components (eg high mannose, complex or hybrid). The high-mannose type N-glycan has five or more mannose residues. The complex-type N-glycan typically has at least one GlcNAc associated with the 1,3-mannose unit and at least one GlcNAc associated with the 1,6-mannose unit of the trimannose core. Complex N-glycans can also have galactose (Gal) or N-acetylgalactosamine (GalNAc) residues, which are optionally modified with sialic acid or derivatives (eg, NANA or NeuAc, where Neu refers to neuraminic acid and Ac refers to acetyl). Complex N-glycans can also have intrachain substitutions containing bisector GlcNAc and core fucose (Fuc). Complex N-glycans can also have multiple antennae on the trimannose core, often referred to as multiple antennal glycans. The hybrid N-glycan has at least one GlcNAc at the end of the 1,3-mannose unit of the trimannose core and zero or more mannose residues at the 1,6-mannose unit of the trimannose core. Various N-glycans are also called glycoforms.

With regard to complex N-glycans, the terms G-2, G-1, G0, G1, G2, A1 and A2 mean the following. G-2 refers to the N-glycan structure, which can be characterized as Man ₃ GlcNAc ₂ ; the term G-1 refers to the structure of the N-glycan, which can be characterized as GlcNAcMan ₃ GlcNAc ₂ ; the term G0 refers to the structure of the N-glycan, which can be characterized as GlcNAc ₂ Man ₃ GlcNAc ₂ ; the term G1 refers to the structure of the N-glycan, which can be characterized as GalGlcNAc ₂ Man ₃ GlcNAc ₂ ; the term G2 refers to the N-glycan structure, which can be characterized as Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ ; the term A1 refers to the structure of the Nglycan, which can be characterized as NANAGal2GlcNAc2Man3GlcNAc2; and the term A2 refers to the N-glycan structure, which can be characterized as NANA ₂ Gal ₂ GlcNAc ₂ Man ₃ GlcNAc ₂ . Unless otherwise indicated, the terms G-2, G-1, G0, G1, G2, A1 and A2 refer to N-glycan types that lack the fucose associated with the GlcNAc residue at the reducing end of the N-glycan. When the term includes F, F indicates that the N-glycan type contains a fucose residue on a GlcNAc residue at the reducing end of the N-glycan. For example, all of G0F, G1F, G2F, A1F, and A2F indicate that the N-glycan further includes a fucose residue linked to a GlcNAc residue at the reducing end of the N-glycan. Lower eukaryotes, such as yeasts and filamentous fungi, do not normally produce N-glycans, which contain fucose.

With respect to multiantennary N-glycans, the term multiantennary N-glycan refers to N-glycans that additionally contain a GlcNAc residue on a mannose residue containing the non-reducing end of the 1,6-unit or 1,3-unit N-glycan, or a GlcNAc residue on each from mannose residues containing the non-reducing end of the 1,6-unit and 1,3-unit of the N-glycan. Thus, multiantennary N-glycans can be characterized by the formulas

GlcNAc (2-4)MangGIcNAc ₂ t Gal (1-4) GlcNAc (2-4)MangGl CNAC2 or NANA(1-4) Gal (1-4) GlcNAc (2-4)MangGlCNAC2

The term 1-4 refers to 1, 2, 3 or 4 residues.

With respect to bisector N-glycans, the term bisector N-glycan refers to N-glycans in which a GlcNAc residue is linked to a mannose residue at the reducing end of the N-glycan. A bisector N-glycan can be characterized by the formula GlcNAc ₃ Man ₃ GlcNAc ₂ where each mannose residue is linked at its non-reducing end to a GlcNAc residue. In contrast, when a multiantennary N-glycan is characterized as GlcNAc ₃ Man ₃ GlcNAc ₂ , this formula indicates that two

- 50 039293 A GlcNAc is linked to a mannose residue at the non-reducing end of one of the two N-glycan units, and one GlcNAc residue is linked to a mannose residue at the non-reducing end of the other N-glycan unit.

Physical properties of the antibody.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also contain one or more glycosylation sites in the variable region of either the light chain or the heavy chain of an immunoglobulin. Such glycosylation sites can lead to increased immunogenicity of the antibody or fragment, or to a change in the pKa of the antibody due to altered antigen binding (Marshall et al. (1972) Annu Rev Biochem 41:673-702; Gala and Morrison (2004) J Immunol 172:5489-94 ; Wallick et al. (1988) J Exp Med 168:1099-109; Spiro (2002) Glycobiology 12:43R-56R; Parekh et al. (1985) Nature 316:452-7; Mimura et al. (2000) Mol Immunol 37:697-706). Glycosylation is known to occur in motifs containing the N-X-S/T sequence.

Each anti-LAG3 antibody (for example, humanized antibodies, such as humanized antagonist antibodies) or antigen-binding fragment (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) have a unique isoelectric point (pI). For example, some antibodies, such as Ab6, have a pI of approximately 6.3.

Each anti-LAG3 antibody (for example, humanized antibodies, such as humanized antagonist antibodies) or antigen-binding fragment (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) have a characteristic melting point, with a higher melting point indicating greater overall in vivo stability (Krishnamurthy R and Manning MC (2002) Curr Pharm Biotechnol 3:361-71). Typically, T _M1 (initial unfolding temperature) may be greater than 60°C, greater than 65°C, or greater than 70°C. The melting point of an antibody or fragment can be measured using differential scanning calorimetry (Chen et al. (2003) Pharm Res 20:1952-60; Ghirlando et al. (1999) Immunol Lett 68:47-52) or circular dichroism (Murray et al (2002) J. Chromatogr Sci 40:343-9).

In a further embodiment, anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) that do not degrade quickly. The degradation of an antibody or fragment can be measured using capillary electrophoresis (CE) and MALDI-MS (Alexander AJ and Hughes DE (1995) Anal Chem 67:3626-32).

In a further embodiment, anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments thereof are selected that have minimal aggregation effects that may result in triggering an unwanted immune response and/or altered or adverse pharmacokinetic properties. Generally, antibodies and fragments with aggregation of 25% or less, 20% or less, 15% or less, 10% or less, or 5% or less are acceptable. Aggregation can be measured in several ways, including size exclusion column (SEC), high performance liquid chromatography (HPLC), and light scattering.

Antibody conjugates.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can be conjugated to the chemical moiety. Such conjugated antibodies and fragments are part of the present invention. The chemical moiety may be, among others, a polymer, a radionuclide or a cytotoxic factor. In specific embodiments, the implementation of the chemical moiety is a polymer that increases the half-life of the antibody or fragment in the body of the individual. Suitable polymers include, but are not limited to, hydrophilic polymers, which include, but are not limited to, polyethylene glycol (PEG) (e.g., PEG with a molecular weight of 2, 5, 10, 12, 20, 30, or 40 kDa), dextran, and monomethoxy polyethylene glycol (mPEG). Lee, et al. (1999) (Bioconj. Chem. 10:973-981) describe PEG-conjugated single chain antibodies. Wen, et al. (2001) (Bioconj. Chem. 12:545-553) describe the conjugation of antibodies to PEG, which is coupled to a radiometal chelator (diethylenetriaminepentaacetic acid (DTPA)).

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can also be conjugated with labels such as ⁹⁹ Tc, ⁹⁰ Y, “Nn, ³² P, ¹⁴ C, ¹²⁵ I, ³ H, ¹³¹ I, 11 C, ¹⁵ O, ¹³ N , ¹⁸ F, ³⁵ S, ⁵¹ Cr, ⁵⁷ To, ²²⁶ Ra, ⁶⁰ Co, ⁵⁹ Fe, ⁵⁷ Se, ¹⁵² Eu, ⁶⁷ CU, ²¹⁷ Ci, ²¹¹ At, ²¹² Pb, ⁴⁷ Sc, ¹⁰⁹ Pd, ²³⁴ Th and ⁴⁰ K, ^157Gd , ^55Mn , ^52Tr , and ^56Fe .

Anti-LAG3 antibodies (for example, humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments described herein (for example,

- 51 039293

4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be PEGylated, eg to increase their biological (eg serum) half-life. To PEGylate an antibody or fragment, the antibody or fragment is typically reacted with a reactive form of polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become bound to the antibody or antibody fragment. In particular embodiments, the PEGylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or similar reactive water-soluble polymer).

As used herein, the term polyethylene glycol encompasses any of the forms of PEG that are used to derivatize other proteins, such as mono-(C1-C ₁₀ )alkoxy or aryloxy polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody or fragment to be pegylated is an aglycosylated antibody or fragment. Methods for PEGylating proteins are known in the art and can be applied to the antibodies of the invention. See, for example, EP 0154316 and EP 0401384.

Anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8 and/or Ab9) can also be conjugated to fluorescent or chemiluminescent labels, including fluorophores such as rare alkaline earth metal chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanate, phycoerythrin, phycocyanin, allophycocyanin, o-phthalaldehyde, fluorescamine, ¹⁵² Eu, dansyl, umbelliferone, luciferin, luminal label, isoluminal label, acridium ester aromatic label, imidazole label, acridinium salt label, oxalate ester label, aequorin, 2,3dihydrophthalazinediones, biotin/avidin, spin labels and stable free radicals.

Anti-LAG3 antibodies (eg, humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments thereof (eg, 4A10, 19E8, 11C9, and/or 22D2; eg, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can also be conjugated to a cytotoxic factor such as diphtheria toxin, Pseudomonas aeruginosa exotoxin A chain, ricin A chain, abrin A chain, modecin A chain, alpha sarcin, Aleurites fordii proteins and compounds (e.g. , fatty acids), dianthine proteins, PAPI, PAPII and PAP-S proteins of Phytoiacca americana, momordica charantia inhibitor, curcin, crotin, saponaria officinalis inhibitor, mitogellin, restrictocin, fenomycin and enomycin.

For conjugation of anti-LAG3 antibodies (e.g. humanized antibodies such as humanized antagonist antibodies) and their antigen-binding fragments (e.g. 4A10, 19E8, 11C9 and/or 22D2; e.g. Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7 , Ab8 and/or Ab9) with different parts, any method known in the art can be used, including those described by Hunter, et al. (1962) Nature 144:945; David, et al. (1974) Biochemistry 13:1014; Pain, et al. (1981) J. Immunol. Meth. 40:219; and Nygren, J. (1982) Histochem. and Cytochem. 30:407. Methods for conjugating antibodies and fragments are common and well known in the art.

Therapeutic applications of anti-LAG3 antibodies.

Also provided are methods for treating or preventing cancer in individuals, such as humans, in need of such treatment by administering an effective amount of anti-LAG3 antibodies or antigen-binding fragments thereof of the present invention as described herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9), which may be effective for such treatment or prevention. In one embodiment, such an individual is suffering from and being treated for a malignant tumor, e.g., a solid tumor, which includes, in addition to tumor cells, tumor-infiltrating lymphocytes (TILs), such as LAG3-expressing T cells, e.g., osteosarcomas, rhabdomyosarcomas , neuroblastoma, kidney cancer, leukemia, transitional cell cancer of the kidney, bladder cancer, Wilms tumor, ovarian cancer, pancreatic cancer, breast cancer (for example, characterized by a mutation in BRCA1 and/or BRCA2), prostate cancer, bone cancer, lung cancer (eg, non-small cell lung cancer), gastric cancer, colorectal cancer, cervical cancer, synovial sarcoma, head and neck cancer, squamous cell carcinoma, multiple myeloma, renal cell carcinoma, retinoblastoma, hepatoblastoma, hepatocellular carcinoma, melanoma , rhabdoid tumor of the kidney, Ewing's sarcoma, chondrosarcoma, malignant brain tumor, glioblastoma, m eningiomas, pituitary adenomas, vestibular schwannoma, primitive neuroectodermal tumor, medulloblastoma, astrocytoma, anaplastic astrocytoma, oligodenroglioma, ependymoma, choroid plexus papilloma, polycythemia vera, thrombocythemia, idiopathic myelofibrosis, soft tissue sarcoma, thyroid cancer, cancerous endometrium, carcinoid liver, breast cancer, or stomach cancer. In one embodiment, the cancer is a metastasis

- 52 039293 a malignant tumor, for example, of the types described above.

The present invention also provides methods for treating or preventing an infectious disease in a subject by administering to the subject an effective amount of the anti-LAG3 antibodies or antigen-binding fragments thereof described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) that may be effective for such treatment or prevention. In one embodiment of the invention, the infectious disease is a viral infection. In one embodiment of the invention, the infectious disease is a bacterial infection. In one embodiment of the invention, the infectious disease is a parasitic infection. In one embodiment of the invention, the infectious disease is a fungal infection.

The present invention provides methods for treating any of the cancers or infectious diseases described herein by administering a therapeutically effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) optionally in association with any other chemotherapeutic agents or therapeutic procedures described in the present invention, as well as compositions comprising such an antibody or fragment together with an additional chemotherapeutic agent.

In one embodiment, the viral infection is an infection with a virus selected from the group consisting of human immunodeficiency virus (HIV), Ebola virus, hepatitis (A, B, or C), herpes virus (e.g., VZV, HSV-I, HAV -6, HSV-II and CMV, Epstein-Barr virus), adenovirus, influenza virus, flaviviruses, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, virus vaccinia, HTLV virus, Dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, or arbovirus encephalitis virus.

In one embodiment of the invention, the bacterial infection is an infection with bacteria selected from the group consisting of Chlamydia, rickettsial bacteria, mycobacteria, staphylococci, streptococci, pneumococci, meningococci and gonococci, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Corynebacterium diphtheriae,

Salmonella, bacilli, Vibrio cholerae, Clostridium tetan, Clostridium botulinum, Bacillus anthricis, Yersinia pestis, Mycobacterium leprae, Mycobacterium lepromatosis and Borriella.

In an aqueous embodiment of the invention, the fungal infection is an infection with a fungus selected from the group consisting of

Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Genus Mucorales (mucor, absidia, rhizopus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and

Histoplasma capsulatum.

In one embodiment of the invention, the parasitic infection is an infection with a parasite selected from the group consisting of Entamoeba histolytica, Balantidium coll,

Naegleria fowleri, Acanthamoeba, Giardia lambia, Cryptosporidium, Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovanl, Toxoplasma gondii, Nippostrongylus brasiliensis.

In addition, the present invention relates to a method for preventing or inhibiting the binding of LAG3 to MHC class II, enhancing the activation of antigen-specific T cells, or stimulating the production of interleukin-2 by T cells in an individual (for example, a human), for example, where the individual suffers from a malignant tumor or an infectious disease (for example, as described herein), comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof (for example, 4A10, 19E8, 11C9, 22D2, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/ or Ab9, to the individual, optionally together with an additional chemotherapeutic agent, eg pembrolizumab or nivolumab.

The scope of the present invention relates to the use of antibodies against LAG3 or their antigennegative fragments described in the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/ or Ab9) for the manufacture of medicinal products

- 53 039293 va for the treatment of a malignant tumor or an infectious disease in an individual.

The present invention relates to methods for treating or preventing osteosarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods of treating or preventing rhabdomyosarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing neuroblastoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing kidney cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, abs, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing leukemia comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing transitional cell cancer of the kidney comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing bladder cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing Wilms' tumor comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing ovarian cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing pancreatic cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods of treating or preventing breast cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment, a method of treating or preventing breast cancer comprises administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention, together with an anthracycline (e.g., doxorubicin and/or epirubicin) and/or a taxane (e.g., paclitaxel and/or docetaxel). ). Optionally, anthracycline and taxane are co-administered with 5-fluorouracil (5FU), cyclophosphamide, and carboplatin. In one embodiment where the breast cancer is HER2 positive, the anti-LAG3 antibody or fragment is co-administered with

- 54 039293 trastuzumab, optionally with taxane and/or pertuzumab.

The present invention relates to methods of treating or preventing prostate cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing bone cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, abs, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing lung cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment of the invention, a method for treating or preventing lung cancer comprises administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention, together with bevacizumab and/or cetuximab.

The present invention relates to methods of treating or preventing non-small cell lung cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment of the invention, a method of treating or preventing non-small cell lung cancer comprises administering an effective amount of an anti-LAG3 antibody or an anti-hex binding fragment thereof of the present invention together with cisplatin, carboplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, gemcitabine, vinorelbine, irinotecan, etoposide, vinblastine and/or pemetrexed.

The present invention provides methods for treating or preventing gastric cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing colorectal cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g. , Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment, a method of treating or preventing colorectal cancer comprises administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention, together with 5-fluorouracil (5-FU), capecitabine, irinotecan, and/or oxaliplatin (e.g., FOLFOX , FOLFIRI, FOLFOXIRI or CapeOx).

The present invention relates to methods of treating or preventing cervical cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods of treating or preventing synovial sarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing head and neck cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing squamous cell carcinoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,

- 55 039293

Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing multiple myeloma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing renal cell carcinoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing retinoblastoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing hepatoblastoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing hepatocellular carcinoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing melanoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , abs, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a rhabdoid tumor of the kidney, comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing Ewing's sarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods of treating or preventing chondrosarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing brain cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing glioblastoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment, a method for treating or preventing glioblastoma multiforme comprises administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention, together with temozolomide.

The present invention relates to methods for treating or preventing meningioma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention.

- 56 039293 (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing pituitary adenoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing vestibular schwannoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a primitive neuroectodermal tumor comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing medulloblastoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing astrocytoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing anaplastic astrocytoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. In one embodiment of the invention, a method of treating or preventing refractory anaplastic astrocytoma comprises administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention together with temozolomide.

The present invention provides methods for treating or preventing oligodenroglioma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing ependymoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing choroid plexus papilloma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing polycythemia vera comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing thrombocythemia comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for the treatment or prevention of idiopathic myelofibrosis, comprising the administration (optionally in conjunction with pembrolizumab or nivolumab)

- 57 039293 an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof according to the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need of it.

The present invention relates to methods of treating or preventing soft tissue sarcoma comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing thyroid cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9, and/or 22D2; e.g., Ab1 , Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing endometrial cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing carcinoid cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing liver cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing breast cancer (e.g., characterized by a mutation in BRCA1 and/or BRCA2) comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10 , 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing gastric cancer comprising administering (optionally in conjunction with pembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing human immunodeficiency virus (HIV) infection in a subject, comprising administering) an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug such as a protease inhibitor, nucleoside/nucleotide reverse transcriptase inhibitor, non-nucleoside reverse transcriptase inhibitors, entry inhibitor, fusion inhibitor, or integrase inhibitors is administered to the individual.

The present invention relates to methods for treating or preventing infection with Bundibugo virus (BDBV), Sudan virus (SUDV), Tai forest virus (TAFV), and/or Ebola virus in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention ( eg 4A10, 19E8, 11C9 and/or 22D2, eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antiviral drug, such as one or more antibodies that specifically bind to BDBV, SUDV, TAFV, or Ebola virus, or a nucleoside inhibitor of RNA polymerase; or a vaccine.

The present invention provides methods for treating or preventing hepatitis A virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antiviral drug.

The present invention relates to methods for treating or preventing hepatitis B virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or

- 58 039293 of its antigen-binding fragment according to the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need of it. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing hepatitis C virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug such as interferon and/or ribavirin is administered to the individual.

The present invention relates to methods for treating or preventing a herpesvirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing a vesicular stomatitis virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a herpes simplex virus I infection in an individual comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing infection with the HAV-6 virus in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a herpes simplex II infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a cytomegalovirus (CMV) infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing an Epstein-Barr virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing an adenovirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing an influenza virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing a flavivirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;

- 59 039293 eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing an echovirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing infection with a rhinovirus in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing Coxsackievirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a coronavirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing respiratory syncytial virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing mumps virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing a rotavirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing measles virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing rubella virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a parvovirus infection in a subject comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing vaccinia virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing human Thlymphotropic Virus (HTLV) infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2 , Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9)

- 60 039293 to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a Dengue virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing papillomavirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a molluscum virus infection in an individual comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing poliovirus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing a rabies virus infection in an individual comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing John Cunningham virus (JC virus) infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, abs, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention relates to methods for treating or preventing an arbovirus encephalitis virus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antiviral drug is administered to the individual.

The present invention provides methods for treating or preventing Chlamydia trachomatis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing infection with rickettsial bacteria in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a mycobacterial infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a staphylococcal infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing streptococcal infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody, or

- 61 039293 antigen-binding fragment of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need in that. Optionally, the subject is administered an antibacterial antibiotic.

The present invention provides methods for treating or preventing pneumococcal infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing meningococcal infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a gonococcal infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a Klebsiella infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention provides methods for treating or preventing a proteus (e.g., P. vulgaris, P. mirabilis, or P. penneri) infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2, eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a serratia infection in a subject comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention provides methods for treating or preventing Pseudomonas infection in a subject by administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5 , Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Legionella infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a Corynebacterium diphtheriae infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention provides methods for treating or preventing Salmonella (e.g., Salmonella bongori or Salmonella enterica) infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19e8, 11C9 and/or 22D2; e.g. , Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing infection with bacilli in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Vibrio cholerae infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or

- 62 039293 antigen-binding fragment of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need in that. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Clostridium tetani infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Clostridium botulinum infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Bacillus anthracis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Yersinia pestis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention provides methods for treating or preventing a Leptospira infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing a Borrelia infection in a subject comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, the subject is administered an antibacterial antibiotic.

The present invention relates to methods for treating or preventing Candida albicans infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Candida krusei infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing a Candida glabrata infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Candida tropicalis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing a Cryptococcus neoformans infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Aspergillus fumigatus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual such as a human,

- 63 039293 who needs it. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Aspergillus niger infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing a Mucorales mucor infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing a Mucorales absidia infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing a Mucorales rhizopus infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Sporothrix schenkii infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing Blastomyces dermatitidis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing a Paracoccidioides brasiliensis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention relates to methods for treating or preventing a Coccidioides immitis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing a Histoplasma capsulatun infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof. Optionally, an antifungal drug is administered to the individual.

The present invention provides methods for treating or preventing an Entamoeba histolytica infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a Balantidium coli infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a Naegleria fowleri infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

- 64 039293

The present invention relates to methods for treating or preventing infection with Acanthamoeba sp. in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need of it.

The present invention provides methods for treating or preventing Giardia lambia infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing infection with Cryptosporidium sp. in an individual, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need of it.

The present invention relates to methods for treating or preventing Pneumocystis carinii infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a Plasmodium vivax infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing infection with Babesia microti in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing a Trypanosoma brucei infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing a Trypanosoma cruzi infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing a Leishmania donovani infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention relates to methods for treating or preventing Toxoplasma gondii infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The present invention provides methods for treating or preventing Nippostrongylus brasiliensis infection in a subject, comprising administering an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) to an individual, such as a human, in need thereof.

The subject is a mammal, such as, for example, a human, a dog, a cat, a horse, a cow, a mouse, a rat, a monkey (eg, a cynomolgus macaque, eg, Masaca fascicularis or Masaca mulatta), or a rabbit.

In specific embodiments, anti-LAG3 antibodies (e.g., humanized antibodies, such as humanized antagonist antibodies) or antigen-binding fragments of the present invention as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9), can be used alone or

- 65 039293 together with other additional drugs and / or therapeutic procedures for the treatment or prevention of any disease, such as a malignant tumor, for example, as described in the present description, in an individual in need of such treatment or prevention. Compositions or kits, for example, pharmaceutical compositions containing a pharmaceutically acceptable carrier containing such antibodies and fragments together with additional drugs, are also part of the present invention.

In specific embodiments, anti-LAG3 antibodies (e.g., a humanized antibody, such as humanized antagonist antibodies) or antigen-binding fragments of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can be used in conjunction with a cancer drug or immunomodulatory drug such as an immunomodulatory receptor inhibitor, for example an antibody or antigen-binding fragment thereof, that specifically binds to a receptor.

In one embodiment, an anti-LAG3 antibody (e.g., humanized antibodies such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are used in conjunction with one or more inhibitors (e.g., a small molecular weight organic compound, or an antibody or antigen-binding fragment thereof) such as: an MTOR inhibitor (mammalian target of rapamycin), a cytotoxic agent, an agent platinum-based, BRAF inhibitor, CDK4/6 inhibitor, EGFR inhibitor, VEGF inhibitor, microtubule stabilizer, taxane, CD20 inhibitor, CD52 inhibitor, CD30 inhibitor, RANK inhibitor (nuclear factor kappa B receptor activator), RANKL inhibitor (receptor activator ligand) nuclear factor kappa B), ERK inhibitor, MAP kinase inhibitor, AKT inhibitor, MEK inhibitor, PI3K inhibitor, HER1 inhibitor, HER2 inhibitor, ing HER3 inhibitor, HER4 inhibitor, Bcl2 inhibitor, CD22 inhibitor, CD79b inhibitor, ErbB2 inhibitor, or farnesyl protein transferase inhibitor.

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are used in conjunction with one or more of: anti-PD1 agents (eg, pembrolizumab, nivolumab, CT-011), anti-PDL1 agents, anti-CTLA4 agents, anti-TIM3 agents, anti- CS1, (eg, elotuzumab), anti-KIR2DL1/2/3 agents (eg, lirilumab), anti-CD27 agents, anti-CD137 agents (eg, urelumab), anti-GITR agents (eg, TRX518), anti-PD-L1 agents (eg. , BMS-936559, MSB0010718C or MPDL3280A), PD-L2 agents, ILT1 agents, ILT2 agents, ILT3 agents, ILT4 agents, ILT5 agents, ILT6 agents, ILT7 agents, ILT8 agents, Anti CD40, funds against OX40, against CD137, against KIR2DL1, against KIR2DL2/3, against KIR2DL4, against KIR2DL5A, against KIR2DL5B, against KIR3DL1, against KIR3DL2, against KIR3DL3, against NKG2A, against NKG2C, against against NKG2E, or any low molecular weight organic inhibitor of such targets; IL-10, anti-IL10 agent, anti-TSLP (thymic stromal lymphoprotein) agent, or pegylated IL-10.

In one embodiment of the invention, the molecular weight of the polyethylene glycol (PEG) portion of the PEGylated IL-10 molecule is about 12,000 daltons, or about 20,000 daltons. In one embodiment, pegylated IL-10 (e.g., human pegylated IL-10) comprises one or more polyethylene glycol molecules covalently linked via a linker (e.g., C _2-12 alkyl such as -CH2CH2CH2-) to one amino acid residue of one subunit IL-10, wherein said amino acid residue is an alpha-amino group of an N-terminal amino acid residue or an epsilon-amino group of a lysine residue. In one embodiment, the pegylated IL-10 is:

(PEG) _b -L-NH-IL-10;

where b is 1-9 and

L is a C _2-12 alkyl linker moiety covalently attached to the nitrogen (N) of one amino acid residue of IL-10.

In one embodiment of the invention, IL-10 of pegylated IL-10 has the formula:

[X--0 (CH ₂ CH ₂ O) _n ] bL-NH-IL-10, where X is H or C _1-4 alkyl;

n is 20-2300;

b is 1-9; and

L is a C _1-n alkyl linker moiety that is covalently attached to the nitrogen (N) of the alpha-amino group at the N-terminus of one IL-10 subunit;

provided that when b is greater than 1, the total number n does not exceed 2300. See US7052686.

- 66 039293

In one embodiment, an anti-IL-10 antibody or antigen-binding fragment thereof (eg, a humanized antibody) contains the CDRs listed below:

CDR-Ll: KTSQNIFENLA (SEQ ID NO: 465)

CDR-L2: NASPLQA (SEQ ID NO: 466)

CDR-L3: HQYYSGYT (SEQ ID NO: 467)

CDR-H1: GFTFSDYHMA (SEQ ID NO: 468)

CDR-H2: SITLDATYTYYRDSVRG (SEQ ID NO: 469)

CDR-H3: HRGFSVWLDY (SEQ ID NO: 470) (see US7662379).

In one embodiment, the anti-TSLP antibody, or antigen-binding fragment thereof (e.g., a humanized antibody), comprises the CDR set as follows:

CDR-Hl: GYIFTDYAMH (SEQ ID NO: 428);

CDR-H2: TFIPLLDTSDYNQNFK (SEQ ID NO: 429);

CDR-H3: MGVTHSYVMDA (SEQ ID NO: 430);

CDR-Ll: RASQPISISVH (SEQ ID NO: 431);

CDR-L2: FASQSIS (SEQ ID NO: 432);

CDR-L3: QQTFSLPYT (SEQ ID NO: 433);

(see WO 2008/76321).

In one embodiment, an anti-CD27 antibody or antigen-binding fragment thereof (e.g., a humanized antibody) comprises a CDR set as follows:

CDR-Hl: GFIIKATYMH (SEQ ID NO: 420);

CDR-H2: RIDPANGETKYDPKFQV (SEQ ID NO: 421);

CDR-H3: YAWYFDV (SEQ ID NO: 422);

CDR-Ll: RASENIYSFLA (SEQ ID NO: 423);

CDR-L2: HAKTLAE (SEQ ID NO: 424);

CDR-L3: QHYYGSPLT (SEQ ID NO: 425);

(see WO 2012/04367).

Thus, the present invention relates to compositions containing an anti-LAG3 antibody or antigen-binding fragment of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) together with pembrolizumab; as well as methods of treating or preventing cancer in a subject, comprising administering to the subject an effective amount of an anti-LAG3 antibody or antigen-binding fragment thereof, together with pembrolizumab (eg, pembrolizumab at a dose of 200 mg every three weeks). Optionally, the subject is also administered in conjunction with another additional drug.

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with the pembrolizumab antibody, which contains an immunoglobulin heavy chain (or its CDR-H1, CDR-H2 and CDR-H3) containing the amino acid sequence:

QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTN

FNEKFKNRVTTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSAST

KGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS

WTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTL

MISRTPEVTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLN

GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAV

EWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL

SLGK (SEQ ID NO: 41); and an immunoglobulin light chain (or its CDR-L1, CDR-L2 and CDR-L3) containing the amino acid sequence:

EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLE

SGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFP

PSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 42).

- 67 039293

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an antibody containing an immunoglobulin heavy chain (or its CDR-H1, CDR-H2 and CDR-H3) containing the amino acid sequence:

QVQLVESGGGWQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRY

YADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 43) ; and an immunoglobulin light chain (or its CDR-L1, CDR-L2 and CDR-L3) containing the amino acid sequence:

EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIP

ARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 44).

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with one or more of: 13-cisretinoic acid, 3-[5-(methylsulfonylpiperadinemethyl)indolyl]quinolone, 4-hydroxy tamoxifen, 5-deoxyuridine, 5'-deoxy -5-fluorouridine, 5-fluorouracil, 6-mercaptopurine, 7hydroxystaurosporine, A-443654, abiraterone acetate, abraxane, ABT-578, acolbifene, ADS-100380, aflibercept, ALT-110, altretamine, amifostine, aminoglutethimide, amrubicine, anagresacridine , anastrozole, angiostatin, AR-23573, ARQ-197, arzoxifene, AS-252424, AS-605240, asparaginase, ATI3387, AT-9263, atrasentan, axitinib, AZD1152, vaccines based on Bacillus Calmette Guerin (BCG), betabulin, BC- 210, besodutox, bevacizumab, BGJ398, bicalutamide a, Bio111, BIO140, BKM120, bleomycin, BMS-214662, BMS-247550, BMS-275291, BMS-310705, bortezimibe, buserelin, busulfan, calcitriol, camptothecin, canertinib, capecitabine, carboplatin, carmustine, CC8490, recombinant CEA vaccine vaccinia-carcinoembryonic antigen), cediranib, CG1521, CG-781, chlamydocin, chlorambucil, chlorotoxin, cilengitide, cimitidine, cisplatin, cladribine, clodronate, cobimetnib, COL-3, CP-724714, cyclophosphamide, cyproterone, cyproterone acetate, cytotarabine, дабрафениба, дакарбазина, дациностата, дактиномицина, далотузумаба, данусертиба, дасатиниба, даунорубицина, декатиниба, дегуелина, денилейкина, дезоксикоформицина, депсипептида, диарилпропионитрила, диэтилстилбестрола, дифтитокса, DNE03, доцетаксела, довитиниба, доксорубицина, дролоксифена, эдотекарина, меченного иттрием-90 эдотреотида, edotreotide, EKB-569, EMD121974, encorafenib, endostatin, enzalutamide, enzastaurin, epirubicin, epithilone B, ERA-923, erbitux, er лотиниба, эстрадиола, эстрамустина, этопозида, эверолимуса, экземестана, фиклатузумаба, финастерида, флавопиридола, флоксуридина, флударабина, флудрокортизона, флуоксиместерона, флутамида, режима FOLFOX, фулвестранта, галетерона, ганетеспиба, гефитиниба, гемцитабина, гиматекана, глюкопиранозиллипида А, гозерелина, гозерелина ацетата , gossypol, GSK461364, GSK690693, HMR-3339, hydroxyprogesterone caproate, hydroxyurea, IC87114, idarubicin, idoxifen, ifosfamide, IM862, imatinib, IMC-1C11, imiquimod, INC280, INCB24360, INO1imba leukipil-2, interferon , иринотекана, JNJ-16241199, кетоконазола, KRX-0402, лапатиниба, лазофоксифена, LEE011, лектрозола, лейковорина, леупролида, леупролида ацетата, левамизола, заключенного в липосомы паклитаксела, ломустина, лонафарниба, люкантона, LY292223, LY292696, LY293646, LY293684, LY294002 , LY317615, LY3009120, marimastat, mechlorethamine, medroxyprogesterone acetate, magestrol acetate, MEK162, melphalan, mercaptop urine, mesna, methotrexate, mithramycin, mitomycin, mitotane, mitoxantrone, suspensions of heat-killed Mycobacterium obuense, tosasertib, MLN8054, natitoclax, neovastat, neratinib, neuradiab, nilotinib, niluthimide, nolatrexed, NVP-BEZ235, oblimersen, otumumide, otumumide, octreotumab Ornatuzumab Orteronel Oxaliplatin Paclitaxel Palbociclib Pamidronate Panitumumab Pazopanib poly-ICLC, porfimer, prednisone, procarbazine, progestins, PSK protein-linked polysaccharide (obtained

- 68 039293 mogo of Basidiomycete coriolus versicolor), PLX8394, PX-866, R-763, raloxifene, raltitrexed, razoxin, ridaforolimus, rituximab, romidepsin, RTA744, rubitecan, scriptaide, Sdx102, seliciclib, selumetinib, selumetinib2, selumetinib SN36093, sorafenibava spironolactone, squalamine, SR13668, streptozocin, SU6668, suberoylanilide hydroxamic acid, sunitinib, synthetic estrogen, talampanel, lagerparepvec talimogen, tamoxifen, temozolomide, temsirolimus, teniposide, tesmilifen, testosterone, testosterone, 6-testosterone, 6-tetrandrine2, tetrandrine2 thiotepa, ticilimumab, tipifarnib, tivozanib, TKI-258, TLK286, TNFa (tumor necrosis factor alpha), topotecan, toremifene citrate, trabectedin, trametinib, trastuzumab, tretinoin, trichostatin A, triciribine phosphate monohydrate, triptorelin pamoate, TSE-424, uracilmustard, valproic acid, valrubicin, vandetanib, vatalanib, VEGF traps, vemurafenib, vinblastine, vincristine, vindesine, vinorel bina, vitaxin, vitespan, vorinostat, VX-745, wortmannin, Xr311, Z-100 hot water extract of Bacillus tuberculosis, zanolimumab, ZK186619, ZK-304709, ZM336372, or ZSTK474.

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with one or more antiemetics, including but not limited to: casopitant (GlaxoSmithKline), netupitant (MGI-Helsinn) and other NK-1 receptor antagonists, palonosetron ( marketed as Aloxi by MGI Pharma), aprepitant (marketed as Emend by Merck and Co.; Rahway, NJ), diphenhydramine (marketed as Benadryl® by Pfizer; New York, NY), hydroxyzine (marketed as Atarax® by Pfizer; New York , NY), metoclopramide (sold as Reglan® by AH Robins Co.; Richmond, VA), lorazepam (sold as Ativan® by Wyeth; Madison, NJ), alprazolam (sold as Xapax® by Pfizer; New York, NY), haloperidol (sold as Haldol® by Ortho-McNe il; Raritan, NJ), droperidol (Inapsine®), dronabinol (sold as Marinol® by Solvay Pharmaceuticals, Inc.; Marietta, GA), dexamethasone (sold as Decadron® by Merck and Co.; Rahway, NJ), methylprednisolone (sold as Medrol® from Pfizer; New York, NY), prochlorperazine (sold as Compazine® from Glaxosmithkline; Research Triangle Park, NC), granisetron (sold as Kytril® from Hoffmann-La Roche Inc.; Nutley, NJ), ondansetron (sold as Zofran® from Glaxosmithkline; Research Triangle Park, NC), dolasetron (sold as Anzemet® from Sanofi-Aventis; New York, NY), tropisetron (sold as Navoban® from Novartis; East Hanover, NJ).

Other side effects of cancer therapy include a deficiency of red blood cells and white blood cells. Thus, in one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an agent that treats or prevents such a deficiency, such as filgrastim, PEG-filgrastim, erythropoietin, epoetin-alpha or darbepoetin-alpha.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with the vaccine. In one embodiment, the vaccine is a cancer vaccine, a peptide vaccine, or a DNA vaccine. For example, in one embodiment of the invention, the vaccine is a tumor cell (eg, an irradiated tumor cell) or a dendritic cell (eg, a dendritic cell treated with a tumor peptide).

In one embodiment, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are administered together with the therapeutic procedure. A therapeutic procedure is one or more steps performed by a physician or clinician in the treatment of an individual that are intended to alleviate one or more symptoms (e.g., cancer and/or infectious disease) in the individual being treated, either by inducing regression or by eliminating such symptoms. symptoms, or by inhibiting the progression of such symptom(s), eg, symptoms of cancer, such as tumor growth or metastasis, to any clinically quantifiable degree.

In one embodiment of the invention, the therapeutic procedure is radiation therapy against a malignant tumor. For example, in one embodiment, the radiation therapy is external beam therapy (EBT): a method of delivering a beam of high energy x-rays to a tumor site. The beam is generated outside the patient (for example, by a linear accelerator) and it is directed to the tumor area. These x-rays can destroy

- 69 039293 malignant cells, and careful planning of treatment allows the preservation of normal surrounding tissues. No radioactive sources are placed inside the patient's body. In one embodiment, the radiation therapy is proton beam therapy: a type of conformal therapy that bombards diseased tissue with protons instead of x-rays. In one embodiment of the invention, the radiation therapy is conformal external beam radiation therapy: a technique that uses advanced technology to tailor radiation therapy to an individual's body structures.

In one embodiment of the invention, radiation therapy is brachytherapy: the temporary placement of radioactive materials into the body, typically used to provide an additional dose or booster dose of radiation to a given area.

In one embodiment of the invention, a surgical procedure performed in conjunction with the administration of an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is a surgical tumorectomy.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an MTOR inhibitor (mammalian target of rapamycin).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a cytotoxic agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a platinum-based agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an EGFR inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a VEGF inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a microtubule stabilizer.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with the CD20 inhibitor taxane.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a CD52 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a CD30 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an inhibitor of RANK (nuclear factor-kappa-B receptor activator).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an inhibitor of RANKL (nuclear factor-kappa-B receptor activator ligand).

- 70 039293

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a BRAF inhibitor, for example in the treatment of melanoma.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a CDK4/6 inhibitor, for example in the treatment of melanoma.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an ERK inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a MAP kinase inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with an AKT inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a MEK inhibitor, for example, for the treatment of melanoma.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a PI3K inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an HER1 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an HER2 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an HER3 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an HER4 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a Bcl2 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a CD22 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a CD79b inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 ,

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Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an ErbB2 inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a farnesyl protein transferase inhibitor.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-PD1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with nivolumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with CT-011.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-PDL1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-CTLA4 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-TIM3 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-CS1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with elotuzumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL1/2/3 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with lirilumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an anti-CD137 antibody or antigen-binding fragment thereof, eg a CD137 agonist antibody or fragment.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with urelumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-GITR agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with TRX518.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-PD-L1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BMS-936559.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with MSB0010718C.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with MPDL3280A.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-PD-L2 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT2 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT3 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT4 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT5 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT6 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT7 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-ILT8 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-CD40 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-OX40 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies)

- 73 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an anti-CD137 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as humanized antagonist antibodies) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL2/3 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL4 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL5A agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR2DL5B agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR3DL1 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR3DL2 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-KIR3DL3 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-NKG2A agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-NKG2C agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-NKG2E agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with IL-10.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an anti-IL10 agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with an anti-TSLP (thymic stromal lymphopoietin) agent.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with pegylated IL-10. In one embodiment, pegylated IL-10 is administered to an individual at a dose of up to 20 μg/kg (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 µg/kg). For example, up to 20 µg/kg is administered every day, eg for up to four (eg 1, 2, 3 or 4) courses of 28 days, eg 20 µg/kg/day for four courses of 28 days .

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with 13-cis-retinoic acid.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with 3-[5-(methylsulfonylpiperadinemethyl)indolyl]quinolone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with 4-hydroxy tamoxifen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with 5-deoxyuridine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with 5'-deoxy-5-fluorouridine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with 5-fluorouracil.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with 6-mercaptopurine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with 7-hydroxystaurosporine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with A-443654.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with abiraterone acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with abraxane.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ABT-578.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with acolbifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ADS-100380.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ALT-110.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with altretamine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with amifostine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with aminoglutethimide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with amrubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with amsacrine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with anagrelide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with anastrozole.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with angiostatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with AP-23573.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ARQ-197.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with arzoxifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with AS-252424.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with AS-605240.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with asparaginase.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody)

- 76 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with AT-9263.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with atrasentan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with axitinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with AZD1152.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with the Bacillus Calmette-Guerin (BCG) vaccine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with batabulin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BC-210.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with besodutox.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with bevacizumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with bicalutamide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with Bio111.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BIO140.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with bleomycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BMS-214662.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BMS-247550.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with BMS-275291.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BMS-310705.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with bortezimib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with buserelin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with busulfan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with calcitriol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with camptothecin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with canertinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with capecitabine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with carboplatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with carmustine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with CC8490.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with CEA (recombinant vaccinia vaccine carcinoembryonic antigen).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with cediranib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with CG-1521.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with CG-781.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with chlamydocin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with chlorambucil.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with chlorotoxin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cilengitide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cimitidine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cisplatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with cladribine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with clodronate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with COL-3.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with CP-724714.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cyclophosphamide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cyproterone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with cyproterone acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with cytarabine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with cytosine arabinoside.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody)

- 79 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used together with dacarbazine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with dacinostat.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with dactinomycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with dalotuzumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with danusertib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with dasatanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with daunorubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with decatanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with deguelin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with denileukin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with deoxycoformycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with depsipeptide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with diarylpropionitrile.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with diethylstilbestrol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with Diftitox.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with docetaxel.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with dovitinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with doxorubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with droloxifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with edotecarin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with yttrium-90 labeled edotreotide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with edotreotide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with EKB-569.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with EMD121974.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with endostatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with enzalutamide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with enzastaurin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with epirubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with epithilone B.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ERA-923.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with cetuximab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody)

- 81 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with erlotinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with estradiol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with estramustine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with etoposide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with everolimus.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with exemestane.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ficlatuzumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with finasteride.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with flavopiridol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with floxuridine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with fludarabine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with fludrocortisone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with fluoxymesterone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with flutamide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with the FOLFOX regimen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a fulvestrant.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with galeterone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with gefitinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with gemcitabine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with gimatecan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with glycopyranosyl lipid A.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with goserelin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with goserelin acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with gossypol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with GSK461364.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with GSK690693.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with HMR-3339.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with hydroxyprogesterone caproate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with hydroxyurea.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with IC87114.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with idarubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody)

- 83 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with idoxifen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with ifosfamide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with IM862.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with imatinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with imiquimod.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with IMC-1C11.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with INCB24360.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with INO1001.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with interferon.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with interleukin-2 (IL-2).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with interleukin-12 (IL-12).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ipilimumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with irinotecan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with JNJ-16241199.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ketoconazole.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with KRX-0402.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with lapatinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with lasofoxifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with lectrozole.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with leucovorin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, and/or Ab9) is used together with leuprolide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with leuprolide acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with levamisole.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with liposomal paclitaxel.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with lomustine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with lonafarnib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with lucanthone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY292223.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY292696.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY293646.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY293684.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with LY294002.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY317615.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with marimastat.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mechlorethamine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with medroxyprogesterone acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with magestrol acetate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with melphalan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mercaptopurine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with mesna.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with methotrexate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mithramycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mitomycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mitotane.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with mitoxantrone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with tosacertib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a suspension of heat-killed Mycobacterium obuense.

- 86 039293

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with MLN8054.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with neovastat.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with neratinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with neuradiab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with nilotinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with niluthimide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with nolatrex.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with NVP-BEZ235.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with oblimersen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with octreotide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ofatumumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with oregovomab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with orteronel.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with oxaliplatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with paclitaxel.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody)

- 87 039293 of the present invention (eg 4A10, 19E8, 11C9 and/or 22D2; eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with palbociclib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with pamidronate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with panitumumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with pazopanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PD0325901.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PD184352.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PEG interferon.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with pemetrexed.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with pentostatin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with perifosine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with phenylalanine mustard.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PI-103.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with pictilisib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PIK-75.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with pipendoxifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PKI-166.

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In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with plicamycin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with poly-ICLC.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a porfimer.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with prednisone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with procarbazine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with progestins.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a PSK protein-bound polysaccharide (obtained from Basidiomycete coriolus versicolor).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PX-866.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with R-763.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with raloxifene.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with raltitrexed.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with razoxin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ridaphorolimus.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with rituximab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with romidepsin.

In one embodiment, the anti-LAG3 antibody or antigen-binding

- 89 039293 fragment (for example, a humanized antibody, such as a humanized antagonist antibody) of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with RTA744.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with rubitecan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with scriptaid.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with Sdx102.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with seliciclib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with selumetinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with semaxanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with SF1126.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with sirolimus.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with SN36093.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with sorafenib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with spironolactone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with squalamine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with SR13668.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with streptozocin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with SU6668.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with suberoylanilide hydroxamic acid.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with sunitinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with synthetic estrogen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with talampanel.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with the talimogen lagerparepvec.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with tamoxifen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with temozolomide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with temsirolimus.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with teniposide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with tesmilifen.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with testosterone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with tetrandrine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with TGX-221.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with thalidomide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with b-thioguanine.

In one embodiment, the anti-LAG3 antibody or antigen-binding

- 91 039293 fragment (for example, a humanized antibody, such as a humanized antagonist antibody) of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with thiotepa.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ticilimumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with tipifarnib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with tivozanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with TKI-258.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with TLK286.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with topotecan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with toremifene citrate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with trabectedin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with trastuzumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with tretinoin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with Trichostatin A.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with triciribine phosphate monohydrate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with triptorelin pamoate.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with TSE-424.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with tumor necrosis factor alpha (TNFa).

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with uracilmustard.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with valproic acid.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with valrubicin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with vandetanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with vatalanib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with a VEGF trap.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with vinblastine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with vincristine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with vindesine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with vinorelbine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with Vitaxin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with vitespan.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with vorinostat.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with VX-745.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with wortmannin.

In one embodiment, the anti-LAG3 antibody or antigen-binding

- 93 039293 fragment (for example, a humanized antibody, such as a humanized antagonist antibody) of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with Xr311.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with zanolimumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with Bacillus tuberculosis extract in hot water Z-100.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ZK186619.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ZK-304709.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ZM336372.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ZSTK474.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with casopitant.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with netupitant.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with an NK-1 receptor antagonist.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with palonosetron.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with aprepitant.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with diphenhydramine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with hydroxyzine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with metoclopramide.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

- 94 039293

Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with lorazepam.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with alprazolam.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with haloperidol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with droperidol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with dronabinol.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with dexamethasone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with methylprednisolone.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with prochlorperazine.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with granisetron.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with ondansetron.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with dolasetron.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with tropisetron.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with filgrastim.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PEG-filgrastim.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used together with erythropoietin.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with epoetin-alpha.

In one embodiment, the anti-LAG3 antibody or antigen-binding

- 95 039293 fragment (for example, a humanized antibody, such as a humanized antagonist antibody) of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with darbepoetin-alpha.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with dabrafenib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with trametinib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with vemurafenib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with cobimetnib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LY3009120.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with DNE03.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with ATI13387.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are co-administered with ganetespib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8, and/or Ab9) are co-administered with encorafenib.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with MEK162.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BKM120.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with LEE011.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with BGJ398.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with INC280.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5,

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Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with PLX8394.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) is used in conjunction with ornatuzumab.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with natitoclax.

In one embodiment, an anti-LAG3 antibody or antigen-binding fragment thereof (e.g., a humanized antibody, such as a humanized antagonist antibody) of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) are used in conjunction with aflibercept.

The term together with indicates that the components that are an anti-LAG3 antibody (for example, a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment of the present invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) together with another agent such as pembrolizumab or nivolumab may be formulated in one composition, e.g. for simultaneous delivery, or may be formulated separately in two or more compositions (for example, a set). Each component can be administered to an individual at a time different from the time of administration of the other component; for example, each administration may be administered non-simultaneously (eg, singly or sequentially) at intervals over a given period of time. Moreover, the individual components can be administered to the individual through the same or different routes (e.g., where the anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or its antigen-binding fragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9 is administered parenterally and paclitaxel is administered orally).

Assays and Experimental and Diagnostic Applications The present invention relates to any method for forming a complex between an anti-LAG3 antibody or antigen-binding fragment of the present invention and LAG3 (e.g., human or cynomolgus LAG3) comprising contacting a LAG3 polypeptide with an anti-LAG3 antibody or fragment in conditions suitable for binding and complex formation.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can be used as affinity purification agents. In this process, anti-LAG3 antibodies and their antigen-binding fragments are immobilized on a solid phase, such as Sephadex, glass or agarose resin, or filter paper, using methods well known in the art. The immobilized antibody or fragment is brought into contact with a sample containing the LAG3 protein (or fragment) to be purified, and the substrate is then washed with a suitable solvent that removes substantially all of the sample material except for the LAG3 protein that has bound to the immobilized antibody or fragment. . Finally, the support is washed with a solvent that elutes bound LAG3 (eg, protein A). Such immobilized antibodies and fragments are part of the present invention.

The present invention relates to methods of using the anti-LAG3 antibodies and antigen-binding fragments of the present invention to determine the degree of T cell activation that a particular individual has or may have in the presence of an antibody or fragment. For example, embodiments of the invention include methods comprising (i) contacting T cells (e.g., CD4+ T cells) from an individual with a superantigen (e.g., any one or more of a staphylococcal superantigen such as SEA, SEB (Staphylococcus enterotoxin B), SEC2, SEC3, SED, SEH, and/or TSST; and/or any one or more of a streptococcal superantigen such as SPE-A, SPE-C, SPE-H, and/or SMEZ-2), e.g., at a concentration of 500 pg /ml or more, such as about 10 ng/ml or 100 ng/ml, in the presence of an anti-LAG3 antibody or fragment (optionally, T cells are preincubated with superantigen (e.g., SEB) and antibody or fragment for about 48 or 72 h) and (ii) determining the level of production of cytokines (eg, TNF-alpha, GM-CSF, IFN-gamma and/or IL-2) by these T cells; where the level of production of the specified cytokine(s) indicates the level of activation of T cells in the presence of the antibody or fragment. Individuals with T cells that exhibit cytokine production (e.g., high levels of cytokine production, such as anti-LAG3 antibody dependent elevation) in the presence of superantigen and antibody are considered the best candidates for antibody or fragment production as a therapy, e.g., for treatment of cancer or infection. In one embodiment of the invention, these best candidates are selected for administration of the antibody or fragment. In one embodiment, such best candidates are administered an effective amount of the antibody or fragment. In one embodiment of the invention, the method includes the step of isolating T cells from the blood of an individual. In one embodiment, the T cells are contacted with an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention and pembrolizumab.

In addition, antigens are provided for the production of secondary antibodies, which are suitable, for example, for Western blotting and other immunoassays described in the present invention. In particular, polypeptides are described that contain the variable regions and/or CDR sequences of an anti-LAG3 antibody or fragment described in the present invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) and which can be used to generate anti-idiotypic antibodies for use in specific detection of the presence of an antibody, for example in the context of therapy.

The present invention relates to cellular ELISA methods using anti-LAG3 antibodies and their antigen-binding fragments of the present invention. In one embodiment of the invention, the method comprises the steps of: (i) contacting cells (e.g., cells or tissue taken from a tumor, e.g., which include lymphocytes putatively expressing LAG3) immobilized on a solid surface (e.g., a microplate) to be tested with respect to the presence of LAG3, with an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention, (ii) optionally washing the mixture to remove unbound anti-LAG3 antibody or fragment, (iii) contacting the anti-LAG3 antibody or fragment with a labeled secondary antibody or its an antigen-binding fragment that binds to an anti-LAG3 antibody or fragment, (iv) optionally washing the complex to remove unbound antibodies or fragments, and (v) detecting the presence of a label on the secondary antibody or fragment; where the detection of the label indicates that the cells contain LAG3. For example, the present invention relates to such cellular ELISA methods for detecting LAG3+ cells in a tumor sample.

The present invention relates to ELISA assays (enzyme-linked immunosorbent assay) comprising the use of an anti-LAG3 antibody (e.g., humanized antibodies such as humanized antagonist antibodies) or an antigen-binding fragment thereof as described herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9).

For example, such a method includes the following steps:

(a) coating the substrate (e.g., the well surface of a microtiter plate, e.g., a plastic plate) with an anti-LAG3 antibody (e.g., humanized antibodies, such as humanized antagonist antibodies) or an antigen-binding fragment thereof (e.g., 4A10, 19E8, 11C9 and/or 22D2, eg Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9);

(b) applying a sample to be tested for the presence of LAG3 to a support (eg, cells taken from a tumor, eg, which include lymphocytes putatively expressing LAG3);

(c) washing the plate so as to remove unbound material in the sample;

(d) applying detectably labeled antibodies (eg enzyme-linked antibodies) that are also specific for the LAG3 antigen;

(e) washing the support to remove unbound labeled antibodies;

(f) if the labeled antibodies are bound to an enzyme, applying a chemical reagent that is converted by the enzyme into a fluorescent signal; and (g) detecting the presence of the labeled antibody.

Detection of a label associated with the substrate indicates the presence of the LAG3 protein. ELISA methods can also be used to identify LAG3+ cells in a tumor sample.

In a further embodiment, the labeled antibody, or its antigen-binding fragment, is labeled with a peroxidase that reacts with ABTS (e.g., 2,2'-azino-bis(3-ethylbenzthiazoline-6sulfonic acid)) or 3,3',5,5'-tetramethylbenzidine, causing a color change that is detectable. Alternatively, the labeled antibody or fragment is labeled with a detectable radioisotope (eg, ³ H) that can be detected using a scintillation counter in the presence of a scintillator.

An anti-LAG3 antibody (for example, humanized antibodies, such as humanized antagonist antibodies) or an antigen-binding fragment thereof according to the invention (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7 , Ab8 and/or Ab9) can be used in Western blotting or immunoblotting of proteins. Such a technique is part of the present invention and includes, for example:

(1) providing a membrane or other solid support containing the sample to be tested for the presence of LAG3, for example, optionally including the step of transferring proteins from the sample to be tested for the presence of LAG3 (for example, after electrophoretic dis-

- 98 039293 separation of proteins in the sample by PAGE or SDS-PAGE), onto a membrane or other solid support using a method known in the art (for example, semi-dry blotting or chamber blotting); and contacting the membrane or other solid support to be tested for the presence of bound LAG3 or fragment thereof with an anti-LAG3 antibody or antigen-binding fragment thereof of the invention.

Such a membrane may be in the form of, for example, a nitrocellulose or vinyl (e.g., polyvinylidene fluoride (PVDF)) based membrane, onto which the proteins to be examined for the presence of LAG3 are transferred in a non-denaturing PAGE (polyacrylamide gel electrophoresis) or SDS-PAGE (polyacrylamide gel electrophoresis) gel. sodium dodecyl sulfate gel electrophoresis) (for example, after gel electrophoretic separation). Before contacting the membrane with the anti-LAG3 antibody or fragment, the membrane is optionally blocked with, for example, skimmed milk powder and the like. for binding non-specific protein binding sites on the membrane;

(2) washing the membrane one or more times to remove unbound anti-LAG3 antibody or fragment and other unbound substances; and (3) detecting bound anti-LAG3 antibody or fragment.

The detection of a bound antibody or fragment indicates that the LAG3 protein is present on the membrane or support and in the sample. Detection of a bound antibody or fragment can be performed by binding the antibody or fragment to a secondary antibody (anti-immunoglobulin antibody) that is labeled with a detectable label, and then detecting the presence of the secondary antibody label.

Anti-LAG3 antibodies (e.g., humanized antibodies such as humanized antagonist antibodies) and antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can also be used for immunohistochemistry. Such a method is part of the present invention and includes, for example, (1) contacting tissue containing TIL and tumor cells (eg, melanoma) to be tested for the presence of a LAG3 protein with an anti-LAG3 antibody or antigen-binding fragment of the invention; and (2) detecting the antibody or fragment on or in the TIL.

If the antibody or fragment is itself labeled with a detectable label, it can be detected directly. Alternatively, the antibody or fragment can be bound with a detectably labeled secondary antibody and then the label detected.

Certain anti-LAG3 antibodies and antigen-binding fragments thereof described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, and/or Ab9) also can be used for tumor imaging in vivo. Such a method may include injecting a detectably labeled, e.g., radioactively labeled, anti-LAG3 antibody or antigen-binding fragment thereof (as described herein) into a patient to be tested for the presence of a tumor associated with LAG3 expression (e.g., which expresses LAG3 , for example, on tumor-infiltrating lymphocytes (TIL)), and then imaging, for example, radionuclide imaging, of the patient's body to detect the presence of a labeled antibody or fragment, for example, in areas containing a high concentration of an antibody or fragment that is associated with or associated with a tumor. Loci detection indicates the presence of LAG3+ TIL in the tumor.

Imaging methods include SPECT (single photon emission computed tomography) imaging or PET (positron emission tomography) imaging. Labels include, for example, iodine-123 ( ¹²³ I) and technetium-99m ( ^99m Tc), for example, in conjunction with SPECT imaging or ^PS , ¹³ N, ¹⁵ O or ¹⁸ F, for example, in conjunction with PET imaging, or indium- 111 (See, for example, Gordon et al. (2005) International Rev. Neurobiol. 67:385-440).

The present invention relates to a method for determining whether a tumor in an individual is responsive to treatment with an anti-LAG3 antibody or antigen-binding fragment of the present invention, comprising determining whether LAG3 is expressed in or on tumor-infiltrating lymphocytes (TIL) and, if said expression is identified, determining that the tumor is responsive to such treatment. Expression of LAG3 by TIL can be determined using any of the methods described in the present invention, for example, ELISA or in vivo imaging. In one embodiment of the invention, the method includes the step of obtaining a sample of said tumor tissue prior to determining LAG3 expression. For example, in one embodiment of the invention, the sample is obtained surgically, for example, by means of a biopsy, for example, a needle biopsy or a partial tumorectomy. In one embodiment, LAG3 expression is determined by contacting TIL with an antibody or fragment and detecting the presence of the antibody or fragment associated with tumor tissue or fragment thereof.

Pharmaceutical compositions and administration.

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For preparing pharmaceutical or sterile compositions of anti-LAG3 antibodies (e.g. humanized antibodies, such as humanized antagonist antibodies) and antigen-binding fragments thereof (e.g. 4A10, 19E8, 11C9 and/or 22D2; e.g. Ab1, Ab2, Ab3, Ab4, Ab5 , Ab6, Ab7, Ab8 and/or Ab9), the antibody or antigen-binding fragment thereof is mixed with a pharmaceutically acceptable carrier or excipient. See, for example, Remington's Pharmaceutical Sciences and U.S. Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984). Such compositions are part of the present invention.

The scope of the present invention relates to dried, eg lyophilized, compositions containing an anti-LAG3 antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, which comprises a pharmaceutically acceptable carrier but is essentially devoid of water.

Therapeutic and diagnostic formulations can be prepared by mixing with suitable carriers, excipients or stabilizers in the form of, for example, lyophilized powders, slurries, aqueous solutions or suspensions (see, for example, Hardman, et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis, et al (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY Lieberman, et al (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY Lieberman, et al (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY).

The present invention relates to any pharmaceutical formulation comprising an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention and PO4 ^2- , e.g. sodium phosphate or potassium phosphate (e.g., about 10 mM), NaCl (e.g., about 7.5 mM), and sucrose (eg, about 3%), for example, having a pH of about 7.4 or about 7.3; or NaOAc (eg, about 20 mM), and sucrose (about 7 or about 9%), for example, having a pH of about 5.0.

The toxicity and therapeutic efficacy of antibody or fragment formulations, administered alone or in combination with another drug, can be determined using standard pharmaceutical techniques in cell cultures or experimental animals, for example, to determine LD ₅₀ (dose lethal to 50% of the population) and ED ₅₀ (dose effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index (LD ₅₀ /ED ₅₀ ). In specific aspects, antibodies exhibiting high therapeutic indices are desirable. The data obtained from these cell culture assays and animal studies can be used in formulating a dosage range for human use. The dosage of such compounds is preferably in the range of circulating concentrations that include the ED ₅₀ with little or no toxicity. The dosage may vary within this range depending on the dosage form used and the route of administration.

In a further embodiment, an additional drug that is co-administered to an individual with an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2 ; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is administered to the individual in accordance with Physicians' Desk Reference 2003 (Thomson Healthcare; 57 ^th edition (November 1, 2002)).

The route of administration may vary. Routes of administration include oral, rectal, transmucosal, intestinal, parenteral; intramuscular, subcutaneous, intradermal, bone marrow, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, dermal, transdermal, or intra-arterial.

The present invention relates to methods for administering an anti-LAG3 antibody, or antigen-binding fragment thereof, comprising administering the antibody or fragment to an individual. For example, the method includes piercing the body of the individual with a syringe needle and injecting the antibody or fragment into the body of the individual, for example, into a vein, artery, tumor, muscle tissue, or under the skin of the individual.

The present invention relates to a container (for example, a plastic or glass container, for example, with a cap, or a chromatographic column, a cannula, or a syringe barrel) containing any of the antibodies or antigen-binding fragments (for example, 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9), polypeptides or polynucleotides described in the present invention, or their pharmaceutical composition containing a pharmaceutically acceptable carrier.

The present invention also relates to an injectable device containing any of the antibodies or antigen-binding fragments, polypeptides or polynucleotides described in this

- 100 039293 of the present invention, or their pharmaceutical composition. An injectable device is a device that delivers a substance to a patient via a parenteral route, such as intramuscular, subcutaneous, or intravenous. For example, the injecting device may be a syringe (for example, pre-filled with a pharmaceutical composition, such as a syringe pen), which, for example, includes a cylinder or barrel containing the liquid to be injected (for example, containing an antibody or fragment or a pharmaceutical composition thereof), a needle for puncturing the skin and/or blood vessels for injecting fluid; and a piston for pushing fluid out of the cylinder and through the cavity of the needle. In one embodiment of the invention, the injection device that contains the antibody or antigen-binding fragment of the present invention, or a pharmaceutical composition thereof, is an intravenous (IV) injection device. Such a device includes an antibody or fragment, or a pharmaceutical composition thereof, in a cannula or trocar/needle that can be connected to a tube that can be connected to a bag or reservoir containing a liquid (e.g., saline; or Ringer's lactate solution containing NaCl). , sodium lactate, KCl, CaCl ₂ and optionally including glucose) administered to the patient through a cannula or trocar/needle. In one embodiment of the invention, the antibody or fragment, or a pharmaceutical composition thereof, may be administered to the device after the trocar or cannula has been placed in the subject's vein and the trocar has been removed from the installed cannula. An IV device may be placed, for example, in a peripheral vein (eg, arm or shoulder); superior vena cava or inferior vena cava, or into the right atrium of the heart (eg, central IV); or into the subclavian, internal carotid or femoral vein, and, for example, can be moved towards the heart until it reaches the superior vena cava or right atrium (eg, central venous line). In one embodiment of the invention, the injection device is a pen; needleless syringe or external infusion pump. A needleless syringe uses a narrow, high-pressure liquid jet that penetrates the epidermis to deliver the antibody or fragment, or pharmaceutical composition thereof, into the patient's body. External infusion pumps are medical devices that deliver an antibody or fragment, or a pharmaceutical composition thereof, to a patient in controlled amounts. External infusion pumps can be driven electrically or mechanically. Different pumps operate in different ways, for example, a syringe pump holds fluid in a syringe reservoir and a movable piston controls fluid delivery, an elastomeric pump holds fluid in an expandable balloon reservoir, and pressure from the balloon's elastic walls delivers fluid. In a peristaltic pump, a set of rollers clamp along the length of the flexible tube, pushing the fluid forward. In a multichannel pump, fluids can be delivered from multiple reservoirs at different rates.

The pharmaceutical compositions described herein can also be administered using a needleless hypodermic device; such as the devices described in US Pat. Nos. 6,620,135; 6096002; 5399163; 5383851; 5312335; 5064413; 4941880; 4,790,824 or 4,596,556. Such needleless devices containing a pharmaceutical composition are also part of the present invention. The pharmaceutical compositions described herein can also be administered by infusion. Examples of well-known implants and modules for administering pharmaceutical compositions include the implants and modules described in US Pat. No. 4,487,603, which describes an implantable microinfusion pump for dispensing a drug at a controlled rate; US Pat. No. 4,447,233, which describes a drug infusion pump for delivering a drug at a specific infusion rate; in US patent No. 4447224, which describes an implantable infusion device with variable flow for continuous drug delivery; US Pat. No. 4,439,196, which describes an osmotic drug delivery system having multi-chamber compartments. Many other such implants, delivery systems and modules are well known to those skilled in the art, and implants, delivery systems and modules containing the pharmaceutical compositions of the present invention are within the scope of the present invention.

Alternatively, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen binding fragment (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) locally rather than systemically, for example by injecting the antibody or fragment directly into the tumor, for example a LAG3+ tumor. Moreover, it is possible to administer the antibody or fragment in a targeted drug delivery system, eg, in a liposome coated with a tissue-specific antibody targeting, eg, a tumor, eg, a LAG3+ tumor, eg, characterized by immunopathology. The liposomes will target and be taken up selectively by the affected tissue. Such methods and liposomes are part of the present invention.

Treating or treating means administering anti-LAG3 antibodies or antigen-binding fragments thereof according to the present invention to an individual having one or more symptoms of a disease in which anti-LAG3 antibodies and antigen-binding fragments thereof are effective.

- 101 039293 useful, for example, in the treatment of an individual having a malignant tumor or an infectious disease, or suspected of having a malignant tumor or an infectious disease, in which the agent has therapeutic activity. Typically, the antibody or fragment is administered in an effective amount or dose that alleviates one or more symptoms (e.g., cancer or infectious disease) in the individual or population being treated, either by inducing regression or elimination of such symptoms, or by inhibiting the progression of such symptoms. symptom(s), for example, symptoms of cancer, such as tumor growth or metastasis, to any clinically measurable degree. The effective amount of the antibody or fragment may vary depending on such factors as the stage of the disease, the age and body weight of the patient, and the ability of the drug to induce the desired response in the individual.

Antibodies (e.g., humanized antibodies, such as humanized antibody antagonists) or antigen-binding fragments thereof as described herein (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may be provided by continuous infusion or by doses administered, for example, every day, 1-7 times a week, once a week, once every two weeks, once a month, once every two months, once a three months, once every six months, annually, etc. Doses may be given, for example, by intravenous, subcutaneous, topical, oral, nasal, rectal, intramuscular, intracerebral, intravertebral, or inhalation routes. An effective dose of an anti-LAG3 antibody or antigen-binding fragment thereof of the present invention is from about 0.1 mg/kg (body weight) to about 100 mg/kg (body weight), for example, for the treatment or prevention of cancer or infectious diseases. In one embodiment of the invention, an effective dose for the treatment of a medical condition in which LAG3 is overexpressed is the dose at which there is complete saturation of the LAG3 antigen in the individual, for example, on T cells in the individual's tumors, or where there is approximately 10% saturation. or more than about 10% saturation.

The determination of an appropriate dose is made by the clinician, for example, using parameters or factors known or believed in the art to provide treatment. As a rule, when determining the dose, the dose begins with an amount slightly less than the optimal dose, and after that it is increased in small increments until the desired or optimal effect is achieved in relation to any negative side effects. Important diagnostic indicators include indicators of symptoms, such as inflammation or the level of inflammatory cytokines produced. It is generally desirable that the biological agent used be from the same species as the animal being treated, thereby minimizing any immune response to the reagent. In the case of a human, for example, chimeric, humanized and fully human antibodies may be desirable. Guidelines for selecting appropriate doses of anti-LAG3 antibodies or fragments thereof are available (see e.g. Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis , Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert et al. (2003) New Engl. J. Med. 348: 601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000 ) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343: 1594-1602).

The presence of mitigation of a disease symptom can be assessed by any clinical measurement commonly used by physicians or other health care providers to assess the severity or state of progression of that symptom. While an embodiment of the present invention (e.g., a method of treatment or article of manufacture) may not be effective in alleviating a given disease symptom(s) in each individual, it should alleviate a given disease symptom(s) in a statistically significant number of individuals when determined using any statistical test known in the art, such as Student's t-test, chi ² test, Mann and Whitney U-test, Kruskal-Wallis (H-test), Jonkheer-Terpstra test, and Wilcoxon test.

Sets.

In addition, kits are contemplated containing one or more components that include, but are not limited to, an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment as described herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) together with one or more additional components, including, but not limited to, an additional drug, such as described in the present description. The antibody or fragment and/or drug can be formulated as a pure composition or in combination with a pharmaceutically acceptable carrier in a pharmaceutical composition.

- 102 039293

In one embodiment, the kit comprises an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment of the invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4 , Ab5, Ab6, Ab7, Ab8 and/or Ab9) or their pharmaceutical composition in one container (for example, in a sterile glass or plastic vial) and an additional drug in another container (for example, in a sterile glass or plastic vial).

In another embodiment, the kit comprises a combination of the invention comprising an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antibody antagonists) or an antigen-binding fragment of the invention (e.g., 4A10, 19E8, 11C9, and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9), or their pharmaceutical composition in combination with one or more drugs formulated together, optionally in a pharmaceutical composition in one common container.

If the kit includes a pharmaceutical composition for parenteral administration to an individual, the kit may include a device for carrying out such administration. For example, the kit may include one or more hypodermic needles or other injectable devices as described above. Thus, the present invention relates to a kit containing an injectable device and an anti-LAG3 antibody or antigen-binding fragment of the present invention, for example, where the injectable device comprises an antibody or fragment, or where the antibody or fragment is in a separate container.

The kit may include a package insert including information regarding the pharmaceutical compositions and dosage forms in the kit. Typically, such information will assist patients and physicians in using the supplied pharmaceutical compositions and dosage forms effectively and safely. For example, the package insert may provide the following information regarding the combination of the invention: pharmacokinetics, pharmacodynamics, clinical trials, efficacy parameters, indications and uses, contraindications, warnings, cautions, adverse reactions, overdose, proper dosage and administration, modes of delivery, proper conditions storage, notes, manufacturer/distributor information and patent information.

Detection Kits and Therapy Kits.

For convenience, an anti-LAG3 antibody (e.g., a humanized antibody such as humanized antagonist antibodies) or an antigen-binding fragment thereof of the invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8 and/or Ab9) may be provided in a set, ie. a packaged combination of reagents in predetermined quantities, with instructions for performing a diagnostic or detection assay. When the antibody or fragment is labeled with an enzyme, the kit includes the substrates and cofactors required by the enzyme (eg, a substrate precursor that provides a detectable chromophore or fluorophore). In addition, other additives such as stabilizers, buffers (eg blocking buffer or lysis buffer) and the like may be included. The relative amounts of the various reagents can vary widely to provide reagent solution concentrations that substantially optimize the sensitivity of the assay. In particular, the reagents may be provided as dry powders, usually lyophilized, including excipients which, when dissolved, provide a reagent solution having the proper concentration.

Also provided are diagnostic reagents and detection kits or detection kits and reagents containing one or more of these reagents for use in various detection assays, including, for example, immunoassays such as ELISA (sandwich-type or competitive format). The components of the kit may be pre-bonded to the solid support, or they can be applied to the surface of the solid support when the kit is in use. In some embodiments, the signal generating agents may be pre-coupled to an antibody or fragment of the invention, or may require combination with one or more components, e.g., buffers, antibody-enzyme conjugates, enzyme substrates, and the like, prior to use. The kits may also include additional reagents, such as blocking reagents to reduce non-specific binding to the surface of the solid phase, washing reagents, enzyme substrates, and the like. The surface of the solid phase may be in the form of a tube, bead, microtiter plate, microsphere, or other materials suitable for immobilizing proteins, peptides, or polypeptides. In specific aspects, an enzyme that catalyzes the formation of a chemiluminescent or chromogenic product or the reduction of a chemiluminescent or chromogenic substrate is a component of the signal generating means. Such enzymes are well known in the art. The kits may contain any of the trapping agents and detection reagents described in the present invention. Optionally, the kit may also contain instructions for performing the methods of the invention.

Also provided is a kit containing an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment thereof,

- 103 039293 packaged in a container, such as a vial or bottle, and further containing a label attached to or packaged with the container, the label describing the contents of the container and providing indications and / or instructions regarding the use of the contents of the container for the treatment of one or more disease states as described in the present description.

In one aspect, the kit is for the treatment of cancer and contains an anti-LAG3 antibody (eg, a humanized antibody, such as a humanized antibody antagonist) or an antigen-binding fragment thereof, and an additional drug or vaccine. The kit may optionally further include a syringe for parenteral, eg intravenous, administration. In another aspect, the kit contains an anti-LAG3 antibody (e.g., a humanized antibody, such as humanized antagonist antibodies) or an antigen-binding fragment thereof, and a label attached to or packaged with a container that describes the use of the antibody or fragment with a vaccine or additional drug. In another aspect, the kit comprises a vaccine or co-drug and a label affixed to or packaged with a container that describes the use of the vaccine or co-drug with the anti-LAG3 antibody or fragment. In certain embodiments, the anti-LAG3 antibody and the vaccine or additional drug are in separate vials, or they are combined together in a single pharmaceutical composition.

As described above in the Combination Therapy section, the co-administration of two drugs does not require that the drugs be administered simultaneously or by the same route, provided that there is an overlap in time during which the drugs demonstrate their therapeutic effect. Simultaneous or sequential administration is envisaged, as well as administration on different days or weeks.

It is also possible to obtain therapeutic kits and detection kits described in the present description, which contain at least one of the antibody, peptide, antigen-binding fragment or polynucleotide described in the present invention, and instructions for using the composition as a detection reagent or drug. Containers for use in such kits typically may contain at least one vial, vial, flask, bottle, syringe, or other suitable container into which one or more detection composition(s) and/or therapeutic composition(s) can be placed. i), and preferably suitably aliquoted. When a second medicament is also provided, the kit may also comprise a second separate container into which the second detection and/or therapeutic composition may be placed. Alternatively, multiple compounds may be prepared in a single pharmaceutical composition and may be packaged in a single container such as a vial, vial, syringe, bottle, or other suitable single container. The kits described herein typically include a means for holding the vial(s) in a commercially restricted space, such as, for example, injection molded or blow molded plastic containers that hold the desired vial(s). When a radioactive label, chromogenic, fluorogenic, or other type of detectable label or detection agent is included in the kit, the labeling agent may either be provided in the same container as the detection or therapeutic composition itself, or alternatively, it may be placed in a second a different container into which this second composition may be placed or suitably aliquoted. Alternatively, the detection reagent or label may be provided in a single container, and in most cases, the kit also typically includes a means to contain the vial(s) in a confined space for commercial sale and/or convenient packaging and delivery.

Also provided is a device or apparatus for carrying out the detection or monitoring methods described in the present invention. Such apparatus may include a chamber or tube into which the sample may be placed, a fluid handling system optionally including valves or pumps to direct sample flow through the device, optional filters to separate plasma or serum from blood, mixing chambers for adding trapping agents or reagents. detection, and optionally a detection device for detecting the amount of detectable label associated with the capture agent immunocomplex. Sample flow can be passive (for example, through capillary, hydrostatic, or other forces that do not require additional manipulation of the device after sample application) or active (for example, using force generated by mechanical pumps, electroosmotic pumps, centrifugal force, or elevated air pressure), or it can be achieved by a combination of active and passive forces.

The following embodiments also provide a processor, a computer-readable memory, and a program stored in the computer-readable memory and adapted to execute on the processor to perform any of the methods described in the present invention. Examples of suitable computer systems, environments and/or configurations include personal computers, server computers, handheld or portable devices, multiprocessor systems, microprocessor-based systems, additional external devices, programmable consumer electronics, network PCs, minicomputers, computers , distributed computing environments that include any of the systems or devices described above, or any other systems known in the art.

General ways.

Standard molecular biology techniques are described in Sambrook, Fritsch and Maniatis (1982 & 1989 2nd ^Edition , 2001 3rd ^Edition ) Molecular Cloning., A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular ^Cloning , 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods are also presented in Ausbel, et al. (2001) Current Protocols in Molecular Biology, Vol. 1-4, John Wiley and Sons, Inc. New York, NY, which describes cloning in bacterial cells and DNA mutagenesis (vol. 1), cloning in mammalian cells and yeast (vol. 2), glycoconjugates and protein expression (vol. 3), and bioinformatics (vol. 4).

Protein purification methods, including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization, are described (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, fusion protein production, protein glycosylation are described (see, e.g., Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York ; Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY, NY, p. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, MO, pp. 45-89 Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, N.J., pp. 384-391). Production, purification and fragmentation of polyclonal and monoclonal antibodies have been described (Coligan, et al. (2001) Current Protcols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard methods for characterizing ligand/receptor interactions are available (see, for example, Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

Monoclonal, polyclonal and humanized antibodies can be prepared (see, for example, Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press, New York, NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 139-243; Carpenter, et al. (2000) J. Immunol. 165: 6205; He, et al. (1998) J. Immunol. 160:1029; Tang et al. (1999) J. Biol. Chem. 274:27371-27378; Baca et al. (1997) J. Biol. Chem. 272:10678-10684; Chothia et al. (1989) Nature 342:877883; Foote and Winter (1992) J. Mol. Biol. 224:487-499; US Pat. No. 6,329,511).

An alternative to humanization is the use of human antibody libraries exposed to phage libraries or human antibody libraries in transgenic mice (Vaughan et al. (1996) Nature Biotechnol. 14:309-314; Barbas (1995) Nature Medicine 1:837-839; Mendez et al (1997) Nature Genetics 15:146-156 Hoogenboom and Chames (2000) Immunol Today 21:371-377 Barbas et al (2001) Phage Display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage Display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397-399).

Single chain antibodies and diabodies have been described (see, for example, Malecki et al. (2002) Proc. Natl. Acad. Sci. USA 99:213-218; Conrath et al. (2001) J. Biol. Chem. 276:7346 -7350; Desmyter et al. (2001) J. Biol. Chem. 276:26285-26290; Hudson and Kortt (1999) J. Immunol. Methods 231:177-189; and U.S. Pat. No. 4,946,778). Bifunctional antibodies have been described (see e.g. Mack, et al. (1995) Proc. Natl. Acad. Sci. USA 92:7021-7025; Carter (2001) J. Immunol. Methods 248:7-15; Volkel, et al (2001) Protein Engineering 14:815-823 Segal, et al (2001) J. Immunol Methods 248:1-6 Brennan, et al (1985) Science 229:81-83 Raso, et al (1997) J Biol Chem 272:27623, Morrison (1985) Science 229:1202-1207, Traunecker, et al (1991) EMBO J 10:3655-3659, and U.S. Patent Nos. 5932448, 5532210 and 6129914).

Bispecific antibodies have also been described (see, for example, Azzoni et al. (1998) J. Immunol. 161:3493; Kita et al. (1999) J. Immunol. 162:6901; Merchant et al. (2000) J. Biol Chem 74:9115 Pandey et al (2000) J Biol Chem 275:38633 Zheng et al (2001) J Biol Chem 276:12999 Propst et al (2000) J Immunol 165:2214; Long (1999) All Rev. Immunol. 17:875).

Purification of the antigen is not necessary to obtain antibodies. Animals can be immunized with cells carrying the antigen of interest. Splenocytes can then be isolated from immunized animals and the splenocytes can be fused to a myeloma cell line to form a hybridoma (see, for example, Meyaard et al. (1997) Immunity 7:283-290; Wright et al. (2000) Immunity 13: 233-242; Preston et al., supra; Kaithamana et al. (1999) J. Immunol. 163:5157-5164).

Antibodies can be conjugated with, for example, small molecule drugs, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic kits, or other purposes, and include antibodies associated with, for example, dyes, radioactive isotopes, enzymes, or metals, such as colloidal gold (see, for example, Le Doussal et al. (1991) J. Immunol 146:169-175 Gibellini et al (1998) J Immunol 160:3891-105 039293

3898; Hsing and Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J. Immunol. 168:883-889).

Flow cytometry techniques, including fluorescence-activated cell sorting (FACS), are available (see, e.g., Owens, et al. (1994) Flow Cytometry Principles for Clinical laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001 ) Flow ^Cytometry , 2nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for nucleic acid modification, including nucleic acid primers and probes, polypeptides and antibodies, for use, for example, as diagnostic reagents, are available (Molecular Probes (2003) Catalogue, Molecular Probes, Inc., Eugene, OR; Sigma-Aldrich (2003) Catalogue, St. Louis, MO).

Standard methods for histology of the immune system are described (see, for example, Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott, Williams, and Wilkins, Phila, PA Louis, et al (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

Software packages and databases for determining, for example, antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments are available (see, for example, GenBank, Vector NTI® Suite (Informax, Inc, Bethesda, MD ); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16: 741-742; Menne, et al. (2000) Bioinformatics Applications Note 16:741-742 Wren, et al (2002) Comput Methods Programs Biomed 68:177-181 von Heijne (1983) Eur J Biochem 133:17-21 von Heijne (1986) Nucleic Acids Res. 14:4683-4690).

Examples

These examples are intended to illustrate the present invention, but not to limit it. The compositions and methods described in the examples are part of the present invention.

Example 1 Obtaining antibodies against human LAG3.

To generate anti-human LAG3 antibodies, Balb/C mice were immunized with 5 μg of labeled recombinant human LAG3 protein-hFc using RIBI adjuvant by footpad injection on a biweekly schedule. Immunized mice were bled and serum titers were determined for binding to human LAG3-transfected CHOK1 cells using cell-based ELISA (described below). Mice with the highest titers were given a final booster immunization with the recombinant protein and the draining popliteal lymph nodes were removed four days later. Hybridomas were generated by electrofusion of isolated lymphocytes with myeloma fusion partner P3X63-AG8.653 using the Cytopulse Hybrimmune electrofusion system. The fused cells were seeded in 96-well plates in DMEM/F12, with 15% BCS, HAT, IL-6, OPI supplement and gentamicin.

Hybridoma supernatants were analyzed for binding to CHOK1 cells expressing human LAG3 and cross-reactivity to CHO cells expressing cynomolgus monkey LAG3 using a cellular ELISA format. CHO-K1 cells expressing human LAG3 and cynomolgus LAG3 were seeded in 96-well tissue culture plates in 50 μl DMEM/F12, with 10% BCS and gentamicin (CHO-Ki medium). Cells were seeded either at 2x10 ⁴ cells/well two days prior to analysis or at 4x10 ⁴ cells/well one day prior to analysis. Before analysis, the medium was removed from the wells and 50 µl of hybridoma supernatant was added. Hybridoma supernatants were incubated for 30-60 min at 37° C. and washed 3 times with PBS/0.05% Tween 20 using a cell ELISA washing protocol on a Biotek EL405x Select CW plate washer. Fifty microliters of HRP-conjugated goat anti-mouse IgG antibody (Southern Biotech cat. no. 103105) at a 1:2000 dilution in CHO-K1 medium was added and incubated at 37°C for 30-60 minutes. Assay plates were washed as described above and developed with TMB and stopped with TMB stop solution (KPL, cat. no. 50-85-06). The absorbance was determined at 450 nm-620 nm.

Positive hybridomas were subcloned by the limiting dilution method or subcloned by hybridoma seeding on semi-solid media and clones were selected on ClonePix (Genetix). The final subclones were grown in small scale cultures in serum free hybridoma production medium and purified using protein G spin columns to obtain a purified antibody for further characterization. Purified antibodies from clones LB145.22D2.E1.1D1 (22D2), LB148.19E8.G1.1A1 (19E8), LB148.4A10.1H1 (4A10), and LB148.11C9.1d (11C9) were further characterized and V _H sequenced. and V.L.

Example 2 Binding of mouse anti-LAG3 antibodies to human LAG3 and cynomolgus macaque LAG3 expressed on the surface of Chinese hamster ovary cells.

Anti-human LAG3 antibodies were examined for binding to CHO-K1 cells expressing human LAG3 and cynomolgus monkey LAG3 using cell-based ELISA. CHO-K1 cells were plated as described above and the medium was removed prior to addition of test samples. Purified antibody from clones LB145.22D2.E1.1D1 (22D2), LB148.19E8.G1.1A1 (19E8),

- 106 039293

LB148.4A10.1H1 (4A10) and LB148.11C9.1C1 (11C9) were serially diluted in DMEM/F12, 10% BCS (CHOK1 medium) and added to CHO-K1 plates. Samples were incubated at 37° C. for 30-60 min and the plates were washed three times with PBS/0.05% Tween-20 using the Biotek cell washer program as described above. Binding detection was performed using a secondary HRP-conjugated goat anti-mouse IgG antibody (Southern Biotech, cat. no. 1031-05) added at a 1:2000 dilution to CHO-K1 medium and incubated at 37°C for 30-60 min. . The assay plates were washed as described above and developed with TMB and stopped with TMB stop solution (KPL, cat. no. 50-85-06). The absorbance was determined at 450 nm-620 nm. Representative binding curves for binding to CHO-K1 cells transfected with human and cynomolgus monkey LAG3 are shown in FIG. one.

Example 3 Determination of affinity for the binding of mouse anti-LAG3 antibodies to recombinant human LAG3 protein.

The binding kinetic activity of mouse clones against human LAG3 LB148.19E8.G1.1A1, LB148.4A1O.1H1, LB148.11C9.1d and LB145.22D2.E1.1D1 using His-tagged recombinant human LAG3 protein was measured by surface plasmon resonance with using the Biacore T200 system (Biacore, GE Healthcare, Piscataway, NJ). Approximately 4,000 RU of a goat antibody specific for the Fc-gamma mouse IgG fragment (Jackson ImmunoResearch, cat. no. 115-006-071, lot 81313) was immobilized by amine addition chemistry on a Series S CM4 sensor chip, cat. no. BR-1005-34 . The mouse anti-human LAG3 clones above were injected over the immobilized surfaces with skin antibodies against mouse proteins at 1 μg/ml for a capture level of 40 RU. HBS-EP+ buffer (BR-1006-69) was used as running buffer at a flow rate of 30 μl/min.

Various concentrations of human LAG3 His protein ranging from 0.15 to 18.8 nM were injected over antibody surfaces at a flow rate of 40 μl/min. After each injection cycle, the surface of the Series S CM4 chip was regenerated using a single injection of 10 mM glycine solution pH 1.5 for 6 s followed by an injection of 12.5 mM NaOH solution at a flow rate of 60 μl/min.

For the analysis of the rate constant of association (k _a ) and dissociation (kd), and the equilibrium dissociation constant K _D used sensograms of binding with background subtraction. The resulting datasets were fitted to a 1:1 Langmuir binding model using Biacore T200 evaluation software (version 2.0). In table. 4 summarizes the affinity for mouse anti-human LAG3 antibodies for recombinant human LAG3.

Table 4

Affinity Measurement of Mouse Anti-Human LAG3 Antibodies for Recombinant Human LAG3

clone ID kal (1/M s) kdl (1/s) K _D (M) LB145.22D2.El .1D1 1.39E+07 3.84Е-05 2.77E-12 LB148.19E8.G1.1A1 7.43E+06 1.09Е-04 1.47E-11 LB148.11С9.1С1 1.31E+06 1.92E-03 1.47Е-09 LB148.4A10.1H1 1.25E+06 1.13E-04 9, OZE-11

Moreover, binding of Ab6 to human CD4, which is structurally similar to LAG3, both of which have four extracellular Ig-like domains, was not detected on BiaCore by FACS when CD4 was expressed on transfected L cells.

Example 4 Effect of Anti-LAG3 Antibodies on 3A9 T-Cell Hybridoma Cells Expressing Human LAG3.

The ability of anti-LAG3 antibodies to enhance antigen-specific T cell activation was examined in a modified version of the previously described T cell activation assay (Workman et al. (2002) Eur. J. Immunol. 32:2255-2263).

Specific to the HEL peptide ₄₈ . A ₆₃ mouse T cell hybridoma (3A9) was challenged with a haplotype matched _MHC11 + treated with HEL peptide _48-63 . B cell line (LK35.2) and IL-2 release were evaluated as an output for antigen specific T cell activation. 3A9 T cell response to HEL-peptide treated ₄₈ . ₆₃ cells LK35.2 was dose dependent.

3A9 T cell lines stably overexpressing mouse or human LAG3 were obtained by retroviral transduction. The present inventors have demonstrated that mouse MHC2 on LK35.2 cells can interact with both human and mouse LAG3 resulting in a marked reduction in antigen-specific IL-2 production, at suboptimal concentrations for T cell activation. The maximum effect of LAG3 activity was observed upon titration of the HELpeptide ₄₈ . ₆₃ at a concentration of 31.2 nM. The inhibitory effect of LAG3 overexpression was not observed when using LK35.2 B cells treated with higher doses of the peptide (corresponding to

- 107 039293 >100 nM). By treatment with 10 μg/ml of a commercially available mouse anti-LAG3 antibody,

C9B7W, the present inventors were able to restore IL-2 to levels of 3A9 cells with the vector.

To evaluate anti-LAG3 antibodies in this assay, 3A9 T cells overexpressing mouse or human LAG3 (100,000 per well) were pretreated with anti-LAG3 antibodies (serially diluted in 3-fold dilutions from 10 μg/mL) for 30 min at 37°C. , and stimulated with LK35.2 cells (33.333/well) treated with 31.2 nM HEL peptide _48-63 for 30 minutes prior to co-culture. After stimulation for 24 h at 37° C. and 5.0% CO ₂ , IL-2 secretion was assessed in culture supernatants at the mesoscale. Inhibition of LAG3 by the antagonist antibody restored T cell function, resulting in restoration of IL-2 production in a dose-dependent manner. IL-2 production was not restored when 3A9 cells were pretreated with isotype control antibodies. The ability of LAG3 overexpression to suppress IL-2 secretion, coupled with the recovery of IL-2 after anti-LAG3 antibody treatment, validates this assay as a reliable screening tool. In table. 5 shows EC ₅₀ for IL-2 recovery using the hLAG3-3A9 mouse anti-human LAG3 antibody system.

Table 5

Mouse anti-human LAG3 antibodies stimulate IL-2 production in the hLAG3-3A9 T cell system

clone ID EC50 (nM) EC50 (nM) LB145.22D2.El .1D1 1, 06 1.65 LB148.19E8.G1.1A1 1.74 1.83 LB148.11С9.1С1 3.56 4.06 LB148.4A10.1H1 2.83 2.96

Example 5 Blocking LAG3/MHC class II binding on Daudi cells.

Mouse clones against human LAG3 were investigated for their ability to block the interaction of hLAG3 with MHC class II. Daudi cells (ATCC #CCL-213) were used as a cell line positive for human MHC class II expression. Daudi cells were blocked with 10 μg/ml goat IgG in HBSS and 2% BCS on ice for 30 min and 0.5 x 106 cells/sample were aliquoted into a 96 well V-buffer plate and the blocking buffer was discarded. Clones LB145.22D2.E1.1D1, LB148.19E8.G1.1A1, LB148.4A1O.1H1 and LB148.11C9.1d were serially diluted starting at 20 µg/mL in HBSS/2% BCS and preincubated with 2 µg/ml human LAG3-huFc in 96-well U-bottomed polypropylene plates in a final volume of 100 µl and incubated on ice for 30 min. After pre-incubation, human-Ec/hybridoma LAG3 supernatant mixtures were added to blocked Daudi cells and incubated for 45 min on ice. Cells were pelleted by centrifugation at 1200 rpm and washed twice with HBSS/2% BCS. Detection of human LAG3-FC binding to Daudi cells was performed using goat anti-human IgG PE conjugate F(ab)' ₂ (Southern Biotech, cat. no.) at a 1:200 dilution in a staining volume of 100 μl and incubated on ice for 20 min. Cells were washed twice as described above, resuspended with HBSS/2% BCS and read on FACSCalibur. In table. 6 summarizes the IC ₅₀ for MHC class II blockade by mouse anti-human LAG3 antibody clones.

Table 6

Mouse anti-human LAG3 antibodies block interaction of human MHC class II with recombinant human LAG3-Fc protein

clone ID 1С ₅₀ (nM) LB145.22D2.El.1D1 2, 10 LB148.19E8.G1.1A1 2.80 LB148.11С9.1С1 2.00 LB148.4A10.1H1 1.90

Example 6 Evaluation of four parental anti-human LAG3 antibodies for tissue cross-reactivity in a defined set of normal human tissues by immunohistochemistry.

Frozen sections of a subgroup of human normal tissues (brain, heart, kidney, liver, lung, pancreas, pituitary) were stained with four anti-human LAG3 antibody clones (LB148.4A10.1H1 (4A10), LB148.11C9.1C1 (11C9) , LB148.19E8.G1.1A1 (19E8), LB145.22D2.E1.1D1 (22D2)) as immunohistochemical reagents to determine potential unexpected tissue reactivity. The human tonsil was used as a positive staining control. Mouse IgG2a, IgG1, and IgG2b, hereinafter referred to as isotype control antibodies, were tested simultaneously with the corresponding anti-human LAG3 clones in all tissues to serve as a comparison in assessing non-specific labeling.

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Immunohistochemical study of cross-reactivity of antibodies 4A10, 11C9, and 19E8 and

22D2 was run in separate assays.

All slides were examined by a pathologist and the immunohistochemical signal was scored on a scale of 0-3 (0=negative, +1=low, +2=moderate, +3=high).

Staining intensity and profile were compared between test article and isotype controls. Test article staining was considered significant when the intensity significantly exceeded that of the isotype control and had a distinct reproducible difference in tissue distribution profile.

Table 7

Clone against Lag3 4A10 (5 µg/ml)

slide# antibody Tissue Identification (Human) Test antibody IgG2aisotype Clone against Lag3 4A10, 5 µg/ml D6414 B1, brain +1 +1 Clone against Lag3 4A10, 5 µg/ml D1106 B1, heart +1 +1 Clone against Lag3 4A10, 5 µg/ml D7462 B1, bud +1 +1 Clone against Lag3 4A10, 5 µg/ml D7122 B1, liver + 2 + 2 Clone against Lag3 4A10, 5 µg/ml D7092 B1, lung +1 +1 Clone against Lag3 4A10, 5 µg/ml D5425 B1, pancreas +2, exocrine cells 0 Clone against Lag3 4A10, 5 µg/ml D5745 B1 pituitary +1 +1 Positive control Clone against Lag3 4A10, 5 µg/ml D7530 B2, tonsil + 2 (corresponding tagging) +1

Positive labeling of pancreatic exocrine cells (approximately 20% of the total population present) is observed only in the sample treated with test antibody, but is limited to the cytoplasm.

Table 8

Clone against Lag3 11C9 (10 µg/ml)

slide# antibody Tissue Identification (Human) Test antibody IgGl- isotype Clone Lag311C9, µg/ml against ten D6414 B1, brain +1 +1 Clone Lag311C9, µg/ml against ten D1106 B1, heart 0 0

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Clone against Lag311C9, 10 µg/ml D7462 B1, bud 0 0 Clone against Lag311C9, 10 µg/ml D7122 B1, liver +1 + 2 Clone against Lag311C9, 10 µg/ml D7092 B1, lung 0 0 Clone against Lag311C9, 10 µg/ml D5425 B1, pancreas 0 0 Clone against Lag311C9, 10 µg/ml D5745 B1, pituitary 0 0 Positive control Clone against Lag311C9, 10 µg/ml D7530 B2, tonsil + 2 (corresponding tagging) 0

All cases of positive signal indicated in the tables (with the exception of positive controls) are interpreted as erroneously associated with the procedure performed, and they are identical to the isotype control or stronger.

Table 9

Clone against Lag3 19E8 (0.5 µg/ml)

slide# antibody Tissue Identification (Human) Test antibody IgG2b isotype Clone against Lag3 D6414 IN 1, +1 (smooth muscle about 19E8, 0.5 µg/ml head brain vessel) Clone against Lag3 D1106 IN 1, +1 (smooth muscle about 19E8, 0.5 µg/ml heart vessel) Clone 19E8, against Lag3 0.5 µg/ml D7462 kidney IN 1, +1 (smooth vessel) muscle +1 Clone against Lag3 D7122 IN 1, +1 (smooth muscle +1 19E8, 0.5 µg/ml liver vessel) Clone 19E8, against Lag3 0.5 µg/ml D7092 light IN 1, 0 +1 Clone 19E8, against Lag3 0.5 µg/ml D5425 B1, pancreas +1 (smooth vessel) muscle 0 Clone against Lag3 D5745 IN 1, +1 (smooth muscle η 19E8, 0.5 µg/ml pituitary vessel) and Positive control Clone 19E8, against Lag3 0.5 µg/ml D7530 B2, tonsil + 3 (corresponding tagging) +1

Non-specific labeling of vascular smooth muscle is observed only in samples treated with test antibody, however it is limited to sarcoplasm/cytoplasm.

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Table 10

Clone against Lag3 22D2 (10 µg/ml)

Antibody Identification tissues (human) Researched antibody IgG2aisotype Clone Lag3 µg/ml against 22D2, 10 D6414 head B1, brain + 2 + 2 Clone Lag3 µg/ml against 22D2, 10 D1106 heart IN 1, +1 +1 Clone Lag3 µg/ml against 22D2, 10 D7462 bud IN 1, + 2 + 2 Clone Lag3 µg/ml against 22D2, 10 D7122 liver IN 1, +1 + 2 Clone Lag3 µg/ml against 22D2, 10 D7092 light IN 1, + 2 + 2 Clone Lag3 µg/ml against 22D2, 10 D5425 B1, pancreas +1 +1 Clone Lag3 µg/ml against 22D2, 10 D5745 pituitary IN 1, + 2 + 2 Positive control Clone Lag3 µg/ml against 22D2, 10 D7530 B2, tonsil +3 (corresponding labeling) + 2

All cases of positive signal indicated in the tables (with the exception of positive controls) are interpreted as erroneously associated with the procedure performed, and they are identical or stronger in the isotype control.

In conclusion, clone 19E8 showed relatively pronounced sarcoplasmic labeling of vascular smooth muscle except in the lung. Clone 4A10 showed cytoplasmic immunohistochemical reactivity in approximately 20% of pancreatic exocrine glandular cells. Clones 11C9 and 22D2 showed labeling in excess of the labeling observed for the isotype control.

Example 7 Binding of anti-Lag3 clones to T cells isolated from human and cynomolgus monkey blood.

Binding of anti-Lag3 mAb clones to primary T cells was evaluated using human and cynomolgus monkey peripheral blood mononuclear cells (PBMCs). Human blood was obtained from local donors; cynomolgus monkey blood was obtained from Bioreclamation. PBMCs were isolated from whole blood using Ficoll-Paque Plus density gradient centrifugation (GE Healthcare, Cat # 17-1440-03) at 524xg for 40 minutes. PBMCs were collected from the media:plasma interface and washed 2 times with phosphate buffered saline (PBS). Residual erythrocytes were lysed using ammonium chloride-potassium erythrocyte lysis solution (ASA, GIBCO, cat. no. A10492-010). Cynomolgus or human PBMC (3×10 ⁶ /ml) were stimulated with 4 μg/ml PHA (Sigma, L2769) for 48 and 20 hours, respectively. For flow cytometric analysis, 1 x 10 ⁶ PHA-activated PBMCs were used for staining in 50 µl of FACS staining buffer (BD, cat. no. 554657). Anti-Lag3 mAb clones or appropriate isotype controls (Table 11) were incubated with human or cynomolgus PBMS and then detected using goat anti-mouse IgG with PE (BD 550589). As phenotypic markers, mouse anti-human CD3 antibody with pacific blue (BD 558124, clone SP34-2), mouse anti-human CD4 antibody with PerCP (BD 550631, clone L200), and mouse anti-human CD8 antibody with APC-Cy7 (BD 557834, clone SK1). Sample preparation was performed on an LSR II flow cytometer and data was analyzed using FlowJo 10.0.6 software (Tree Star, Inc.).

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Flow cytometric analysis revealed the binding of all analyzed clones against

Lag3 with primary human and cynomolgus macaque T cells (representative data are shown in Fig. 2).

Table 11

Clones against Lag 3 and isotype controls used in the study

TS31.ZE1.S7 28ACM m!gG2a/K 10 mM Na phosphate, 75 mM NaCl, 3% sucrose, pH 7.3 ТС31.27F11.С2 64AFW mlgGl/K 10 mM phosphate, 75 mM NaCl, 3% sucrose, pH 7.4 KM9.LP1.MAVZ 63ADP mIgG2b/K 20 mM Na acetate, 7% sucrose, pH 5.5 Own Lag3 Lot No. Isotype Compound LB148.19Е8.Gl.ΙΑ one 42AGE m!gG2b/K 10 mM PO4, 75 mM NaCl, 3% sucrose, pH 7.4 LB145.22D2.El . ID one 98 AGE m!gG2a/K 10 mM PO4, 75 mM NaCl, 3% sucrose, pH 7.4 LB148.4A10.1H1 45AGE m!gG2a/K 10 mM PO4, 75 mM NaCl, 3% sucrose, pH 7.4 LB148.11C9.1C1 47AGE mlgGl/K 20 mM NaAc, 9% sucrose, pH 5.0

Example 8 Epitope mapping of an anti-hLAG3 antibody by hydrogen-deuterium exchange mass spectrometry.

Contacts between antibodies against human LAG3 and LAG3 were determined using hydrogen-deuterium exchange mass spectrometric analysis.

Materials.

Human LAG3-His:[VEGF leader]-LAG3_H-[His9G] (LQPGAEVPWWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPH

PAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRA

AVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPV RESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSR VGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQ LNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQL LSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLHHHHHHHHHGGQ) (SEQ ID NO: 447)

Anti-hLAG3 clone 22D2 (chimeric antibody containing [LAG3_H] mAb (LB145.22D2.E1.D1 (VHO/VLO) Geneart) IgG4 S228P/kappa (PX) and a human IgG4 framework containing the hinge-stabilizing mutation S228P).

Clone 11C9 against hLAG3 ([LAG3_H] mAb (LB148.11C9.1d) mouse IgG1/kappa (HY)/mouse IgG1).

Clone 4A10 against hLAG3 ([LAG3_H] mAb (LB148.4A10.1H1) mouse IgG2a/kappa (HY)/mouse IgG2a).

Liquid chromatography-mass spectrometry.

The mass spectrometer was a Thermo Scientific Orbitrap-Velos. For measurements in deuterium-labeled samples, the mass spectrometer was set up to acquire one complete set of orbittrap MS scan data at a resolution power of 60,000, a target ion count of 1E6, and a maximum ion injection time of 500 ms. To obtain MS/MS data for peptide identification, the mass spectrometer was set up to acquire one full scan spectrum at a resolution power of 60,000 followed by ten data-dependent MS/MS spectra in the ion trap.

The liquid chromatography system was a Waters nanoAcquity for analytical column gradient and a Waters 515 isocratic pump for digestion and sample loading. For sample digestion and loading, the buffer used was 2% acetonitrile and 0.05% trifluoroacetic acid at a flow rate of 80 μl/min. For the analytical gradient, the buffers were buffer A) 0.1% formic acid in water and buffer B) 0.1% formic acid in acetonitrile.

The gradient was 40 µl/min from 2% B to 36% B over 10 min, followed by a wash with 80% B for 1.5 min and re-equilibration in 2% B for 3 min. The column was then washed with a cycling gradient of 2 to 80% B, three times with a time of 1 min each step, followed by a final equilibration at 2% B for 5 minutes. The capture column was a Waters Vanguard C18 BEN, 1.7 µm guard column and the analytical column was a Waters C18 BEN300, 1.7 µm 1x50 mm column.

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Sample processing for deuterium labeling was performed using a Leaptec H/D-X PAL system. The labeling tray was set to 25°C, the quench tray was set to 1.5°C, and the capture column chamber and analytical column were set to

1.5°C. An immobilized pepsin column (Porosyme Immobilized Pepsin, 2.1x30 mm, Life Technologies) was kept outside the column chamber at room temperature.

Labeling with deuterium.

For antibody clones 22D2 and 11C9, hLAG3-His (113 pmol/µl) was mixed with an equal volume of antibody (55 pmol/µl) or, as an unbound control, PBS, pH 7.6. The antibody-bound samples and unbound control were incubated at room temperature for 1 hour before starting the labeling experiment. For 4A10, antibody (30 pmol/µl) was mixed with hLAG3-His in a 2:1 v/v ratio. The subsequent steps were identical for all samples.

To label the samples with deuterium, 2 µl sample was mixed with 25 µl PBS in deuterium oxide, pD 7.6. The labeling times were 30, 300, 3000 or 9000 s. After the specified time, 25 μl of the labeling mixture was added to 40 μl of cold quench buffer (8 M urea, 100 mM TCEP, 0.02% dodecyl maltoside). The quenched sample was incubated at 1.5°C for 2 min. Then 55 µl was injected into the cooling chamber of the column, where the sample passed through the pepsin column and the resulting peptides were loaded into the capture column. After 3 min, the analytical gradient and mass spectrometer were started.

A fully deuterated sample was obtained by incubating 2 μl hLAG3 with 108 μl deuterated denaturing buffer (4 M urea, 100 mM TCEP, 0.01% DDM in 99.5% deuterium oxide). The sample was incubated at 37°C overnight. 55 µl was directly injected into the column chamber and data was obtained as before.

Data analysis.

LC-MS/MS data were generated from an unlabeled sample and searched prior to deuterium labeling to confirm successful protein digestion and to obtain a list of peptides. The data was searched in the database using the Proteome Discoverer 1.4 search algorithm and SEQUEST HT (ThermoFisher Scientific). The protein database used was the human-His LAG3 sequence linked to the Saccharomyces cerevisiae yeast database.

MS data after labeling with deuterium was processed using HDExaminer (Sierra Analytics). The weight and retention time selected by the software for each peptide was manually confirmed.

Results.

Human LAG3 residues protected by antibodies 22D2, 4A10 and 11C9 when bound to the LAG3 protein are illustrated in FIG. 3, 4(a-c) and 5(a-b).

Example 9 Cellular KinExA.

The affinity of anti-LAG3 antibodies for LAG3 was determined using a cellular kinetic exclusion assay (KinExA). Cellular KinExA can be used to measure the affinity of a molecule for a cell surface binding partner (Rathanaswami et al. Analytical Biochemistry 373(1): 52-60 (2008); Xie et al. J. Immunol. Methods 304 (1-2) : 1-14 (2005)). In this case, BaF/3 cells were stably transfected with human and cynomolgus monkey LAG3 proteins. Transfected cells or parental control BaF/3 cell line were grown to a density of 1.7x106 to 3.2x106 cells per ml at 37°C, 120 rpm, 5% CO ₂ , in 1x RPMI 1640 medium with 10% FBS, 10 ng/ml IL-3, 5 µg/ml puromycin. Cells were concentrated, mixed with 15 pM or 150 pM antibody in cell culture medium, and incubated for 24 to 48 hours at room temperature with rotation at 20 to 30 rpm. Cells were present at a maximum concentration of 2x107 cells per ml (parental BaF/3 or cynomolgus macaque LAG3 transformants) or 1x107 cells per ml (human LAG3 transfectants) and were diluted in a 2-fold series of 18 representatives. Cells were pelleted and free antibody in the supernatant was quantified using a KinExA 3200 device (Sapidyne, Idaho, USA). The device coupled the free antibody to polymethyl methacrylate beads (Sapidyne) that were coated with goat anti-human Fey F(ab') ₂ (Jackson ImmunoResearch Laboratories, Pennsylvania, USA). The antibody on the beads was labeled with 1.5 μg/ml Alexa Fluor® 647 conjugated goat anti-(Fab') ₂ antibody (Jackson ImmunoResearch Laboratories), washed, and fluorescent signal read using KinExA™ 3200. Data for each antibody concentration of 15 and 150 pM was subjected to simultaneous fit using KinExA™ Pro n-Curve Analysis software version 4.0.11 (Sapidyne).

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Table 12

KinExA affinity measurement

K _D for human LAG3 K _D for LAG3 cynomolgus macaque K _D (pM) η K _D (pM) η Chimera 22D2 3 2 eleven 2 VH6/VL3 Hu22D2 (Ab 5) 6 2 25 2 VH6/VL3 N55S Hu22D2 (Ab 7) ten 2 eleven 2 VH6/VL3 N55Q Hu22D2 (Ab 8) eleven 2 16 2 VH6/VL3 N55D Hu22D2 (Ab 6) 2 3 12 3

Example 10 Effect of Anti-LAG3 Antibodies on 3A9 Mouse T-Cell Hybridoma Cells Expressing Human LAG3.

The ability of anti-LAG3 antibodies to enhance antigen-specific T cell activation was examined in a modified version of the previously described T cell activation assay (Workman et al., Eur. J. Immunol. 32:2255-2263 (2002)).

HEL peptide _{48-63 specific mouse} T cell hybridoma (3A9) was stimulated with haplotype matched MHC11+ treated with HEL peptide _48-63 B cell line (LK35.2) and IL-2 release was assessed as an output for antigen-specific activation T cells. The 3A9 T cell response to HEL peptide-treated _{48-63 LK35.2} cells was dose dependent.

3A9 T cell lines stably overexpressing mouse or human LAG3 were obtained by retroviral transduction. It has been demonstrated that mouse MHC2 on LK35.2 cells can interact with both human and mouse LAG3 resulting in a marked reduction in antigen-specific IL-2 production at suboptimal concentrations for T cell activation. The maximum effect of LAG3 activity was observed upon titration of the HEL peptide _48-63 at a concentration of 31.2 nM. An inhibitory effect of LAG3 overexpression was not observed with LK35.2 B cells treated with higher doses of the peptide (corresponding to >100 nM). By treatment with 10 μg/ml of a commercially available mouse anti-LAG3 antibody, C9B7W, IL-2 levels were restored to levels in 3A9 cells with the vector.

To assess the effect of anti-LAG3 antibodies in this assay, 3A9 T cells overexpressing mouse or human LAG3 (100,000 per well) were pretreated with anti-LAG3 antibodies (serially diluted in 3-fold dilutions from 10 µg/mL) for 30 min at 37°C. C, and stimulated with LK35.2 cells (33.333/well) treated with 31.2 nM HEL peptide _48-63 for 30 minutes prior to coculture. After stimulation for 24 h at 37° C. and 5.0% CO ₂ , IL2 secretion was assessed in culture supernatants at the mesoscale. Inhibition of LAG3 by the antagonist antibody restored T cell function, resulting in restoration of IL-2 production in a dose-dependent manner. IL-2 production was not restored when 3A9 cells were pre-treated with isotype control antibodies. The ability of LAG3 overexpression to suppress IL-2 secretion coupled with IL-2 recovery following anti-LAG3 antibody treatment validates this assay as a reliable screening tool. In table. 13 shows EC ₅₀ for IL-2 recovery using the hLAG3-3A9 mouse anti-human LAG3 antibody system.

Table 13

Mouse anti-human LAG3 antibodies stimulate IL-2 production in the hLAG3-3A9 T cell system

Antibody EC ₅₀ range, nM (n) LB145.22D2. El.1D1 1.06-1.65 (2) LB148.19E8.G1.1A1 1.74-1.83 (2) LB148.11С9.1С1 3.56-4.06 (2) LB148.4A10.1H1 2.83-2.96 (2) Chimera 22D2 0.69-1.91 (5) Hu22D2 VH6/VL3 (Ab 5) 0.57-1.07 (6) Hu22D2 VH6/VL3 N55S (7\b 7) 0.45-1.27 (6) Hu22D2 VH6/VL3 N55D (Ab 6) 0.47-1.01 (6) Hu22D2 VH6/VL3 N55Q (Ab 8) 0.72-1.08 (6)

Example 11 Blocking LAG3/MHC class II binding on Daudi cells.

Mouse anti-LAG3 human clones and humanized anti-LAG3 clones were investigated for their ability to block the interaction of hLAG3 with human MHC class II. Daudi cells (ATCC #CCL-213) were used as a cell line positive for human MHC class II expression. Daudi cells were blocked with 10 μg/ml goat IgG in HBSS and 2% BCS on ice during those

- 114 039293 30 min and 0.5 x ^{10 6} cells/sample were aliquoted into a 96-well V-shaped buffer plate and the blocking buffer was removed. Clones LB145.22D2.E1.1D1, LB148.19E8.G1.1A1, LB148.4A1O.1H1, and LB148.11C9.1d and hu22D2 VH6/VL3, hu22D2 VH6/VL3 N55Q, hu22D2 VH6/VL3 N55S, hu22D2 VH VL3 N55D and chimeric 22D2 were serially diluted starting at 20 µg/mL in HBSS/2% BCS and pre-incubated with 2 µg/mL human-huFc LAG3 or biotinylated human-huFc LAG3 in 96-well U-shaped polypropylene plates. bottomed in a final volume of 100 µl and incubated on ice for 30 min. After pre-incubation, human LAG3-Fc/antibody supernatant mixtures were added to blocked Daudi cells and incubated for 45 min on ice. Cells were pelleted by centrifugation at 1200 rpm and washed twice with HBSS/2% BCS. Detection of human LAG3-FC binding to Daudi cells was performed using goat anti-human IgG PE conjugate F(ab)' ₂ (Southern Biotech, cat. no.) at a 1:200 dilution of unconjugated huLAG3-huFc or at a 1:500 dilution of streptavidin -PE for biotinylated huLAG3-huFc in a staining volume of 100 µl and incubated on ice for 20 min. Cells were washed twice as described above, resuspended with HBSS/2% BCS and read on FACSCalibur. In table. 14 summarizes the IC ₅₀ for MHC class II blockade by mouse anti-human LAG3 antibody clones.

Humanized anti-human LAG3 antibodies (VH6/VL3, VH6/VL3 N55D, VH6/VL3 N55Q, VH6/VL3 N55S and chimeric 22D2) were tested for their ability to block the interaction of hLAG3 with human MHC class II as described above. Biotinylated human LAG3-huFc was used and detection was performed using streptavidin-PE.

Table 14 Mouse anti-human LAG3 antibodies block interaction of human MHC class II with recombinant . protein LAG3 human-F c

Antibody IC50( _hM ) LB145.22D2.El .1D1 2.1 LB148.19E8.G1.1A1 2.8 LB148.11С9.1С1 2.0 LB148.4A10.1H1 1.9 Chimeric 22D2 2.5 Hu22D2 VH6/VL3 (Ab 5) 2, 6 Hu22D2 VH6/VL3 N55S (AJo 7) 2.1 Hu22D2 VH6/VL3 N55D (AJo 6) 2.4 Hu22D2 VH6/VL3 N55Q (AJo 8) 2.5

Example 12 Binding of anti-Lag3 clones to T cells isolated from human and cynomolgus monkey blood.

Binding of anti-LAG3 mAb clones to primary T cells was evaluated using human and cynomolgus monkey (cyno) peripheral blood mononuclear cells (PBMCs). Human blood was obtained from local donors; cynomolgus monkey blood was obtained from Bioreclamation. PBMCs were isolated from whole blood using Ficoll-Paque Plus density gradient centrifugation (GE Healthcare, Cat # 17-1440-03) at 524xg for 40 minutes. PBMCs were collected from the media:plasma interface and washed 2 times with phosphate buffered saline (PBS). Residual erythrocytes were lysed using ammonium chloride-potassium erythrocyte lysis solution (ASA, GIBCO, cat. no. A10492-010). Cynomolgus or human PBMC (3×10 ⁶ /ml) were stimulated with 4 μg/ml PHA (Sigma, L2769) for 48 and 20 hours, respectively. For flow cytometric analysis, 1x10 ⁶ PHA-activated PBMCs were used for staining in 50 µl of FACS staining buffer (BD, cat. no. 554657). Anti-Lag3 mAb clones or appropriate isotype controls were incubated with human or cynomolgus PBMS and then detected using goat anti-mouse IgG with PE (BD 550589). As phenotypic markers, mouse anti-human CD3 antibody with pacific blue (BD 558124, clone SP34-2), mouse anti-human CD4 antibody with PerCP (BD 550631, clone L200), and mouse anti-human CD8 antibody with APC-Cy7 (BD 557834, clone SK1). Sample preparation was performed on an LSR II flow cytometer and data was analyzed using FlowJo 10.0.6 software (Tree Star, Inc.).

Flow cytometric analysis revealed binding to all analyzed anti-LAG3 clones to primary human and cynomolgus T cells. The EC _50s for binding human and cynomolgus CD4+ and CD8+ T cells were determined for humanized 22D2 antibodies and are summarized in Table 1. 15. Data correspond to 3 human donors and 3 cynomolgus monkey donors after stimulation with 4 μg/ml PHA for 40 h before staining.

Table 15

EC ₅₀ for binding to human and cynomolgus monkey CD4+ and CD8+ T cells

(EC50, nM) LAG-3 people LAG-3 macaque Javanese CD4 ⁺ CD8 ⁺ CD4 ⁺ CD8 ⁺ Hu22D2 7) VH6/VL3 N55S (ab 57 49 35 41 Hu22D2 6) VH6/VL3 N55D (ab 39 33 thirty thirty Hu22D2 eight) VH6/VL3 N55Q (ab 41 35 27 31

Example 13 Effect of anti-human LAG-3 antibody treatment +/- anti-PD-1 treatment on IL-2 production in SEB stimulated human PBMCs.

Primary T Cell Assays - The first assay examined LAG3 blocking function alone or in combination with an anti-PD-1 agent in relation to increased IL-2 production by T cells after activation by human PBMC SEB (FIG. 7). Neutralization of LAG3 activity, alone and in the presence of an anti-PD-1 agent, resulted in increased production of IL-2. Examples of mesoscale amounts of IL-2 from 2 donors in dose titration of anti-LAG3 antibody are given. Hu22D2-enhanced IL-2 production by SEB-stimulated PBMC 1.54-fold (range 1.15-2.36-fold, n=8) compared to isotype control and in combination with anti-PD1 agent 1.45-fold (range 1.15-2.36 times, n=4 at 10 or 0.3 µg/mL anti-PD-1 agent) compared to anti-PD-1 agent plus isotype control, Hu22D2 alone or in combination with PD-1 agent, demonstrated comparable activity to a reference anti-LAG3 antibody. Hu2D2 is Ab6.

a-PD-1 is a fully human anti-PD1 Ig64 monoclonal antibody. The reference anti-LAG3 antibody is a fully human anti-human LAG3 monoclonal IgG4 antibody that binds to an additional loop of human LAG3.

Example 14 Effect of hu22D2 +/- anti-PD-1 treatment on IFN-γ and TNFa production in MLR stimulated human PBMCs.

The MLR system was also used to study hu22D2 activity in primary T cells. Human PBMCs are stimulated with monocyte-derived dendritic cells from a different donor, resulting in T cell activation due to MHC mismatch. In this analysis, a high degree of variability in donor responses was observed. However, preliminary data demonstrated that neutralization of LAG3 activity, alone and in the presence of anti-PD-1 antibody, enhanced T cell activation in two of three donors as measured by IFN-γ and TNF-α production (FIG. 8). Hu2D2 is Ab6.

Example 15 Pharmacokinetic and pharmacodynamic analysis of anti-LAG3 antibody in cynomolgus monkeys.

The pharmacokinetic and pharmacodynamic profile of Ab6 was evaluated in cynomolgus monkeys. Methods involving the housing and use of animals in the study were reviewed and approved. During the study, the care and use of animals was carried out in accordance with the principles described in the guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC), Animal Welfare Act, the American Veterinary Medical Association (AVMA) Euthanasia Panel on Euthanasia, and the Institute for Laboratory Animal Research (ILAR) Guide to the Care and Use of Laboratory Animals.

25 naive male cynomolgus monkeys of Chinese origin (4.0-7.0 kg at the time of dosing) were used. The animals were observed twice a day. In addition, the observation of the animals was carried out at each time point of blood sampling. Body weight was recorded once before each dosing event.

In the study, 5 males were assigned to each of the 5 groups. Animals in all groups received 5 doses of test or control products iv via the cephalic vein over 10 minutes. Doses for groups were 0.03, 0.3, 1, 10 and 30 mg/kg. Samples for pharmacokinetic studies were taken from all animals on day 1: before the dose and after 15 minutes and 1, 4, 8, 24 (day 2), 48 (day 3) and 96 (day 5) hours after the dose. Samples were also collected on Day 8: pre-dose and 1, 8, 24 (Day 9), 48, (Day 10), 120 (Day 13), and 168 (Day 15) hours post-dose. All samples were processed to obtain plasma and stored frozen at 70±10°C until analysis. Taking samples for the study of pharmacodynamics was carried out for all animals on day 1: before the dose and after 15 minutes and 1, 4, 8, 24 hours (day 2), 48 hours (day 3) and 96 hours (day 5) after the dose . Taking samples

- 116 039293 were also carried out on day 8: before the dose and after 1, 8, 24 hours (day 9), 48 hours (day 10), 120 hours (day 13) and 168 hours (day 15) after the dose. All samples were processed to obtain plasma and stored frozen at -70±10°C until analysis.

Total antibody and free (unbound antibody and partially bound) were evaluated using two different assays: universal assay and antigen capture assay, respectively. This evaluation is shown in Fig. 9 (results of the Ag capture assay) and FIG. 10 (results of universal analysis). Steady state clearance and volumetric distribution for each dose level were evaluated and are shown in Table 1. 16 (antigen capture assay) and 17 (universal assay). Differences between clearance and volumetric distribution parameters of the antibody demonstrated that a portion of the AL6 antibody interacted with its target in cynomolgus monkeys.

Table 16

LAG3 antigen capture assay - AL6 clearance and volume distribution at steady state (Vss) at various doses

Dose (mg/kg) Clearance (ml/h/kg) Vss (ml/kg) 0.03 6, 8 76.4 0.3 0.48 75.5 one 0.45 95.3 ten 0.33 72.3 thirty 0.29 63.6

The antigen capture data revealed free antibody in the sample. These antigen uptake data (data not shown) showed a 23-fold difference in clearance over the studied dose range (0.003-30 mg/kg) (Table 16).

Table 17

Total Antibody Analysis - Ab6 Clearance and Steady State Volume Distribution (Vss) at Different Doses

Dose (mg/kg) Clearance (ml/h/kg) Vss (ml/kg) 0.03 0.94 47 0.3 0.36 48.9 one 0.33 56.1 ten 0.26 60.9 thirty 0.26 66.5

Total antibody data detected a total antibody in the sample. This total antibody data (FIG. 9) showed a 3.6-fold difference in clearance over the studied dose range (0.003-30 mg/kg) (Table 17).

The total concentration of AL6 in cynomolgus monkeys was assessed at doses of 0.03, 0.3, 1, 10 and 30 mg/kg over time. This evaluation is shown in Fig. 10. As the dose was increased, antibody clearance decreased in individuals. Dose-normalized AL6 concentration over time is shown in FIG. 11. At later time points after the second IV bolus of AL6, there was a decrease in exposure indicating potential immunogenicity against the antibody.

LAG3 target-mediated clearance of AL6 antibody was observed when the concentration of antibody in monkeys was examined over time. To describe the concentration-time profiles for AL6, a two-compartment PK model was obtained with linear and non-linear (Vmax, Km) clearance parameters. These data are presented in Fig. 12.

All references cited in this specification are incorporated herein by reference as if it were specifically and individually indicated that each individual publication, database entry (e.g., Genbank sequence or GenelD entry), patent application, or patent is included in as links. Applicants imply that the statement of incorporation under 37 C.F.R. 1.57(b)(1), refers to each individual publication, database entry (e.g., Genbank sequence or GenelD entry), patent application, or patent, each explicitly identified under 37 C.F.R. 1.57(b)(2), even if such a citation is not immediately adjacent to a special inclusion statement. The inclusion of specific inclusion by reference statements, if any, in the specification does not in any way detract from that general inclusion statement. The citation of references in this specification is not intended to imply that the reference is prior art, nor is it intended to be an admission as to the content or date of these publications or documents.

The scope of the present invention is not limited to the specific embodiments described in the present invention. Indeed, various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art.

- 117039293 from the previous description and the accompanying drawings. Such modifications are intended to be within the scope of the appended claims.

The above description is considered sufficient to enable a person skilled in the art to carry out the invention in practice. Various modifications of the invention, in addition to those shown and described in the present description, will become apparent to those skilled in the art from the above description and are within the scope of the appended claims.

Claims (20)

1. An antibody or antigen-binding fragment that specifically binds to human LAG3, comprising: (1) a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35); or (2) a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35); or (3) a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35); or (4) a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35); or (5) a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); - 118 039293 CDR-H2, which contains the amino acid sequence DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35). 2. An antibody or antigen-binding fragment according to claim 1, including a light chain variable domain containing CDR-L1 which contains the amino acid sequence KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2, which contains the amino acid sequence GASNLES (SEQ ID NO: 39); and CDR-L3, which contains the amino acid sequence QQSTEDPRT (SEQ ID NO: 40); and a heavy chain variable domain containing CDR-H1, which contains the amino acid sequence DYNVD (SEQ ID NO: 33); CDR-H2, which contains the amino acid sequence DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3 which contains the amino acid sequence NYRWFGAMDH (SEQ ID NO: 35). 3. An antibody or antigen-binding fragment according to claim 1, including: (1) a light chain variable domain containing the amino acid sequence of amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain containing the amino acid sequence of amino acids 1-119 of SEQ ID NO: 106; or (2) a light chain variable domain comprising the amino acid sequence of amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain comprising the amino acid sequence of amino acids 1-119 of SEQ ID NO: 108; or (3) a light chain variable domain comprising the amino acid sequence of amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain comprising the amino acid sequence of amino acids 1-119 of SEQ ID NO: 110; or (4) a light chain variable domain comprising the amino acid sequence of amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain comprising the amino acid sequence of amino acids 1-119 of SEQ ID NO: 112; or (5) a light chain variable domain comprising the amino acid sequence of amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain comprising the amino acid sequence of amino acids 1-119 of SEQ ID NO: 122. 4. An antibody or antigen-binding fragment according to claim 1, including: (1) an immunoglobulin light chain containing the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain containing the amino acid sequence of SEQ ID NO: 106; or (2) an immunoglobulin light chain comprising the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 108; or (3) an immunoglobulin light chain comprising the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 110; or (4) an immunoglobulin light chain comprising the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 112; or (5) an immunoglobulin light chain comprising the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 114; or (6) an immunoglobulin light chain comprising the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 116; or (7) an immunoglobulin light chain comprising an amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and - 119 039293 immunoglobulin heavy chain containing the amino acid sequence of SEQ ID NO: 118; or (8) an immunoglobulin light chain containing the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126 and an immunoglobulin heavy chain containing the amino acid sequence of SEQ ID NO: 120; or (9) an immunoglobulin light chain containing the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and an immunoglobulin heavy chain containing the amino acid sequence of SEQ ID NO: 122. 5. An antibody or antigen-binding fragment according to any one of claims 1-4, which is humanized. 6. An antibody or antigen-binding fragment that specifically binds to human LAG3 that contains a light chain variable domain containing amino acids 21-131 of SEQ ID NO: 126 and a heavy chain variable domain containing amino acids 1-119 of SEQ ID NO: 116. 7. An antibody or antigen-binding fragment according to claim 6, which contains an immunoglobulin light chain containing the amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and an immunoglobulin heavy chain containing the amino acid sequence of SEQ ID NO: 116. 8. An antibody that specifically binds to human LAG3, which consists of two immunoglobulin heavy chains and two immunoglobulin light chains, where each immunoglobulin light chain contains an amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and each immunoglobulin heavy chain contains an amino acid a sequence of amino acids 1-119 of SEQ ID NO: 116, and a human IgG4 constant region. 9. An antibody that specifically binds to human LAG3, which consists of two immunoglobulin heavy chains and two immunoglobulin light chains, where each immunoglobulin light chain contains an amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and each immunoglobulin heavy chain contains an amino acid the sequence of SEQ ID NO: 116. 10. An antibody that specifically binds to human LAG3, which consists of two immunoglobulin heavy chains and two immunoglobulin light chains, where each immunoglobulin light chain consists of an amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and each immunoglobulin heavy chain consists of from the amino acid sequence of SEQ ID NO: 116. 11. An antibody that specifically binds to human LAG3, which consists of two immunoglobulin heavy chains and two immunoglobulin light chains, where each immunoglobulin light chain consists of an amino acid sequence of amino acids 21-238 of SEQ ID NO: 126, and each immunoglobulin heavy chain contains amino acid sequence from amino acids 1-119 of SEQ ID NO: 116. 12. An antibody or antigen-binding fragment according to any one of claims 1-11, which is glycosylated with one or more CHO N-linked glycans selected from the group consisting of G0-F, G1F, G2-F, G0, G1, G2 and Man5. 13. An antibody or antigen-binding fragment according to any one of claims 1 to 11, which is expressed in Chinese hamster ovary cells. 14. A polynucleotide encoding the light and heavy chain of an antibody or antigen-binding fragment according to claims 1-11. 15. An expression vector containing a polynucleotide according to claim 14. 16. A pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of claims 1-13 and an anti-PD-1 antibody. 17. Pharmaceutical composition containing an antibody according to claim 2 and pembrolizumab. 18. Pharmaceutical composition containing an antibody according to claim 9 and pembrolizumab. 19. A pharmaceutical composition containing an antibody according to claim 10 and pembrolizumab. 20. A pharmaceutical composition containing an antibody according to claim 11 and pembrolizumab.