Antibody that specifically binds human tim-3, pharmaceutical composition and use of such antibody or composition to enhance t cell activation or treat cancer

Abstract

This disclosure provides antibodies that bind specifically to TIM-3 (e.g., human TIM-3) and antagonize the function of TIM-3. Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for producing these antibodies, and methods of treating an individual using these antibodies are also provided.

Description

“ANTIBODY THAT SPECIFICALLY BINDS TO HUMAN TIM-3, PHARMACEUTICAL COMPOSITION AND USE OF SAID ANTIBODY OR COMPOSITION TO ENHANCE T CELL ACTIVATION OR TREAT CANCER” 1. CROSS-REFERENCE TO RELATED ORDERS

[001] This application claims the benefit of U.S. Provisional Applications Nos. 62 / 342,610, filed May 27, 2016; and 62 / 420,276, filed November 10, 2016, each of which is incorporated by reference in its entirety. 2. FIELD

[002] This disclosure relates to antibodies that bind specifically to TIM-3 (e.g., human TIM-3) and methods for using them. 3. FUNDAMENTALS

[003] T cell mucin and immunoglobulin domain 3 (TIM-3) protein is a type I membrane protein in the immunoglobulin (Ig) superfamily. It has an extracellular Ig (IgV) variable-like domain, an extracellular mucin-like domain, and a cytoplasmic domain with six conserved tyrosine residues (Monney et al. (2002) Nature 415:536-541). TIM-3 is expressed on activated type 1 helper T lymphocytes (Th1) and CD8+ T lymphocytes (Tc1), some macrophages (Monney et al. (2002) Nature 415:536-541), activated natural killer (NK) cells (Ndhlovu et al. (2012) Blood 119(16):3734-3743), and IL-17-producing Th17 cells (Nakae et al. (2007) J Leukoc Biol 81: 1258-1268).

[004] Studies have shown that TIM-3 functions to inhibit T cell, myeloid cell, and NK cell-mediated responses and promote immune tolerance. For example, TIM-3 IgV peptide fused with an immunoglobulin domain, which binds to and neutralizes TIM-3 ligands, caused hyperproliferation of Th1 cells and release of Th1 cytokines in immunized mice (Sabatos et al. (2003) Nat Immunol Petition 870260065483, dated 03 / 07 / 2026, page 12 / 214 2 / 195 4:1102-1110). In fact, in vivo administration of an anti-TIM-3 antibody intensified the pathological severity of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis (Monney et al. (2002) Nature 415:536-541). Furthermore, TIM-3 expression is upregulated in CD8+ T cells in cancer patients. For example, approximately 30% of NY-ESO-1 specific CD8+ T cells in patients with advanced melanoma exhibit upregulation of TIM-3 expression (Fourcade et al. (2010) J Exp Med 207:2175-2186).

[005] Given the apparent role of human TIM-3 in modulating immune responses, therapeutic agents designed to antagonize TIM-3 signaling represent great promise for the treatment of diseases involving TIM-3-mediated immune suppression. SUMMARY

[006] This disclosure provides antibodies that specifically bind to TIM-3 (e.g., human TIM-3) and antagonize TIM-3 function, e.g., TIM-3-mediated immune suppression. Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for producing these antibodies, and methods of treating an individual using these antibodies are also provided. The antibodies disclosed herein are particularly useful for increasing T cell activation in response to an antigen (e.g., a tumor antigen or an infectious disease antigen) and / or decreasing Treg-mediated immune suppression and, therefore, for treating cancer in an individual or treating or preventing an infectious disease in an individual.

[007] Consequently, in one aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising CDRH1, CDRH2 and CDRH3 complementarity-determining regions and a variable light chain region comprising CDRH1, CDRH2 and CDRH3 complementarity-determining regions Petition 870260065483, dated 03 / 07 / 2026, p. 13 / 214 3 / 195 of complementarity CDRL1, CDRL2 and CDRL3, where: (a) CDRH1 comprises the amino acid sequence X1X2X3X4X5S (SEQ ID NO: 48), where Xi is R, S, A, G, K, M, or T. X2 is Q, S, A, G, R, or T. X3 is N, Y, G, or Q. X4 is A or Q, and X5 is W, M, A, S, or T; (b) CDRH2 comprises the amino acid sequence of WVSAISGSGGSTY (SEQ ID NO: 2); (c) CDRH3 comprises the amino acid sequence of AKGGDYGGNYFD (SEQ ID NO: 3); (d) CDRL1 comprises the amino acid sequence of X1ASQSVX2SSYLA (SEQ ID NO: 52), where X1 is R or G; and X2 is missing or is S; (e) CDRL2 comprises the amino acid sequence of X1ASX2RAT ​​(SEQ ID NO: 53), where X1 is D or G; and X2 is N, S, or T; and (f) CDRL3 comprises the amino acid sequence of QQYGSSPX1T (SEQ ID NO: 54), where X1 is L or I.

[008] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region comprising CDRH1, CDRH2 and CDRH3 complementarity-determining regions and a variable light chain region comprising CDRL1, CDRL2 complementarity-determining regions Petition 870260065483, dated 03 / 07 / 2026, p. 14 / 214 4 / 195 and CDRL3, where: (a) CDRH1 comprises the amino acid sequence X1X2X3X4X5S (SEQ ID NO: 48), where Xi is R, S, A, G, K, M, or T. X2 is Q, S, A, G, R, or T. X3 is N, Y, G, or Q. X4 is A or Q, and X5 is W, M, A, S, or T; (b) CDRH2 comprises the amino acid sequence of WVSAISGSGGSTY (SEQ ID NO: 2); (c) CDRH3 comprises the amino acid sequence of AKGGDYGGNYFD (SEQ ID NO: 3); (d) CDRL1 comprises the amino acid sequence of X1ASQSVX2SSYLA (SEQ ID NO: 52), where X1 is R or G; and X2 is missing or is S; (e) CDRL2 comprises the amino acid sequence of X1ASX2RAT ​​(SEQ ID NO: 53), where X1 is D or G; and X2 is N, S, or T; and (f) CDRL3 comprises the amino acid sequence of QQYGSSPX1T (SEQ ID NO: 54), where X1 is L or I.

[009] In another aspect, the present invention provides an antibody or isolated antibody comprising a variable heavy chain region having CDRH1, CDRH2 and CDRH3 complementarity-determining regions and a variable light chain region having CDRL1, CDRL2 and CDRL3 complementarity-determining regions, wherein the antibody is internalized after binding to cells that Petition 870260065483, dated 03 / 07 / 2026, page 15 / 214 5 / 195 express human TIM-3, where CDRH3 comprises the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 3).

[010] In another aspect, the present invention provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region having complementarity-determining regions CDRH1, CDRH2 and CDRH3 and a variable light chain region having complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein the antibody is internalized after binding to cells expressing human TIM-3, and wherein CDRH3 comprises the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 3).

[011] In certain modalities: (a) CDRH1 comprises the amino acid sequence X1X2X3X4X5S (SEQ ID NO: 48), where X1 is R, S, A, G, K, M, or T. X2 is Q, S, A, G, R, or T. X3 is N, Y, G, or Q. X4 is A or Q, and X5 is W, M, A, S, or T; (b) CDRH2 comprises the amino acid sequence of WVSAISGSGGSTY (SEQ ID NO: 2); (c) CDRL1 comprises the amino acid sequence of X1ASQSVX2SSYLA (SEQ ID NO: 52), where X1 is R or G; and X2 is missing or is S; (d) CDRL2 comprises the amino acid sequence of X1ASX2RAT ​​(SEQ ID NO: 53), where X1 is D or G; and Petition 870260065483, dated 03 / 07 / 2026, p. 16 / 214 6 / 195 X2 is N, S, or T; and (e) CDRL3 comprises the amino acid sequence of QQYGSSPXiT (SEQ ID NO: 54), where Xi is L or I.

[012] In certain embodiments, CDRH1 comprises the amino acid sequence X1X2NAWS (SEQ ID NO: 49), where: X1 is R or A; and X2 is Q or R. In certain embodiments, CDRH1 comprises the amino acid sequence X1X2GQX3S (SEQ ID NO: 50), where: X1 is K, M, or G; X2 is A or S; and X3 is S or T. In certain embodiments, CDRH1 comprises the amino acid sequence X1X2QQAS (SEQ ID NO: 51), where: X1 is S, R, T, or G; and X2 is A, S, T, or G. In certain embodiments, CDRH1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 and 4 to 12.

[013] In certain embodiments, CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 13 to 16. In certain embodiments, CDRL2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 21. In certain embodiments, CDRL3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 and 23.

[014] In certain embodiments, CDRH1, CDRH2 and CDRH3 comprise the amino acid sequences CDRH1, CDRH2 and CDRH3, respectively, shown in the SEQ ID NOs: 1, 2 and 3; 4, 2 and 3; 5, 2 and 3; 6, 2 and 3; 7, 2 and 3; 8, 2 and 3; 9, 2 and 3; 10, 2 and 3; 11, 2 and 3; or 12, 2 and 3.

[015] In certain embodiments, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences CDRL1, CDRL2 and CDRL3, respectively, shown in the SEQ ID NOs: 13, 17 and 22; 14, 17 and 22; 15, 18 and 22; 14, 19 and 22; 14, 20 and 22; 14, 21 and 22; 16, 20 and 22; or 14, 17 and 23.

[016] In certain embodiments, CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences shown in the SEQ ID NOs: Petition 870260065483, dated 03 / 07 / 2026, p. 17 / 214 7 / 195 1, 2, 3, 14, 21 and 22; 4, 2, 3, 14, 21 and 22; 5, 2, 3, 14, 21 and 22; 6, 2, 3, 14, 21 and 22; 7, 2, 3, 14, 21 and 22; 8, 2, 3, 14, 21 and 22; 9, 2, 3, 14, 21 and 22; 10, 2, 3, 14, 21 and 22; 11, 2, 3, 14, 21 and 22; or 12, 2, 3, 14, 21 and 22, respectively.

[017] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 1, 2, 3, 14, 21 and 22, respectively.

[018] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3 comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 1, 2, 3, 14, 21 and 22, respectively.

[019] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 5, 2, 3, 14, 21 and 22, respectively.

[020] In another aspect, the present disclosure provides an antibody or anti Petition 870260065483, dated 03 / 07 / 2026, page 18 / 214 8 / 195 isolated body that specifically binds to human TIM-3 comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 5, 2, 3, 14, 21 and 22, respectively.

[021] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 9, 2, 3, 14, 21 and 22, respectively.

[022] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3 comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 9, 2, 3, 14, 21 and 22, respectively.

[023] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, Petition 870260065483, dated 03 / 07 / 2026, page 19 / 214 9 / 195 CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 1, 2, 3, 15, 18, and 22, respectively.

[024] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3 comprising a variable heavy chain region comprising complementarity-determining regions CDRH1, CDRH2 and CDRH3, and a variable light chain region comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise the amino acid sequences presented in SEQ ID NOs: 1, 2, 3, 15, 18 and 22, respectively.

[025] In certain modalities, the antibody is internalized after binding to cells that express human TIM-3.

[026] In another aspect, the present invention provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody is internalized after binding to cells expressing human TIM-3.

[027] In certain embodiments, a smaller percentage of cells expressing human TIM-3 survive in the presence of the antibody than in the presence of pab1944w (IgG1 N297A) in an assay comprising the following steps: (a) placing cells in plates expressing human TIM-3 at 2 x 104 cells per well in a tissue culture plate; (b) adding 1111 ng / ml of αHFc-NC-DM1 and 1111 ng / ml of the antibody or pab1944w (IgG1 N297A) to a final volume of 100 μl / well; (c) incubating at 37°C and 5% CO2 for 72 hours; (d) measuring the survival of cells expressing human TIM-3; and (e) calculating the percentage of survival relative to untreated cells expressing human TIM-3. In certain modes, the cell survival rate in the presence of the antibody is at least 50% lower than the cell survival rate in the presence of pab1944w (IgG1 N297A). In certain modes, cells expressing TIM-3 Petition 870260065483, dated 03 / 07 / 2026, page 20 / 214 10 / 195 human TIM-3 cells are Kasumi-3 cells. In certain embodiments, the cells expressing human TIM-3 are Kasumi-3 cells (ATCC® CRL-2725™). In certain embodiments, the cells expressing human TIM-3 are Jurkat cells genetically modified to express human TIM-3.

[028] In certain embodiments, a smaller percentage of cells expressing human TIM-3 survive in the presence of the antibody than in the presence of Hum 11 (IgG4 S228P) in an assay comprising the following steps: (a) placing cells in plates expressing human TIM-3 at 2 x 104 cells per well in a tissue culture plate; (b) adding 1111 ng / ml of αHFc-NC-DM1 and 1111 ng / ml of antibody or Hum 11 (IgG4 S228P) to a final volume of 100 μl / well; (c) incubating at 37°C and 5% CO2 for 72 hours; (d) measuring the survival of cells expressing human TIM-3; and (e) calculating the percentage of survival relative to untreated cells expressing human TIM-3. In certain modalities, the cell survival rate in the presence of the antibody is at least 50% lower than the cell survival rate in the presence of Hum11 (IgG4 S228P). In certain modalities, cells expressing human TIM3 are Kasumi-3 cells.In certain embodiments, the cells expressing human TIM-3 are Kasumi-3 cells (ATCC® CRL-2725™). In certain embodiments, the cells expressing human TIM-3 are Jurkat cells genetically modified to express human TIM-3.

[029] In certain embodiments, the antibody comprises a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 55. In certain embodiments, the antibody comprises a variable heavy chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 35. In certain embodiments, the variable heavy chain region comprises an amino acid sequence selected from the Petition 870260065483, dated 03 / 07 / 2026, page 21 / 214 11 / 195 group consisting of SEQ ID NOs: 24 to 35. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 25. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 28. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 32. In certain embodiments, the N-terminal glutamate (E) residue of a variable heavy chain region of an antibody as described in this document is replaced by a pyroglutamate (pE) residue.

[030] In certain embodiments, the antibody comprises a variable light chain region comprising the amino acid sequence of SEQ ID NO: 56. In certain embodiments, the antibody comprises a variable light chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 36 to 47. In certain embodiments, the variable light chain region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 36 to 47. In certain embodiments, the variable light chain region comprises the amino acid sequence of SEQ ID NO: 46. In certain embodiments, the N-terminal glutamate (E) residue of a variable light chain region of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[031] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 35. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 25. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 28. In certain embodiments, the variable heavy chain region comprises Petition 870260065483, dated 03 / 07 / 2026, p. 22 / 214 12 / 195 the amino acid sequence of SEQ ID NO: 32. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 58. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 65. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 70, 71, 72, 73, 74 or 75. In certain embodiments, the N-terminal glutamate (E) residue of a heavy chain of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[032] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 35. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 25. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 28. In certain embodiments, the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 32. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 58. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61.In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 65. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 70, 71, 72, 73, 74, or 75. Petition 870260065483, dated 03 / 07 / 2026, p. 23 / 214 13 / 195

[033] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable light chain region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 36 to 47. In certain embodiments, the variable light chain region comprises the amino acid sequence of SEQ ID NO: 46. In certain embodiments, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 76 or 77. In certain embodiments, the N-terminal glutamate (E) residue of a light chain of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[034] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable light chain region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 36 to 47. In certain embodiments, the variable light chain region comprises the amino acid sequence of SEQ ID NO: 46. In certain embodiments, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 76 or 77.

[035] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region and a variable light chain region, wherein the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 25 and 46; 28 and 46; 29 and 46; 30 and 46; Petition 870260065483, dated 03 / 07 / 2026, page 24 / 214 14 / 195 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46. In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 25 and 46. In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 28 and 46. In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 32 and 46. In certain embodiments, the N-terminal glutamate (E) residue of a variable heavy chain region of an antibody as described in this document is replaced by a pyroglutamate (pE) residue and / or the N-terminal glutamate (E) residue of a variable light chain region of the antibody is replaced by a pyroglutamate (pE) residue.

[036] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a variable heavy chain region and a variable light chain region, wherein the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in the SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46. In certain embodiments, the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences shown in SEQ ID NOs: 25 and 46.In certain embodiments, the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in SEQ ID Nos: 28 and 46. In certain embodiments, the amino acid sequences Petition 870260065483, dated 03 / 07 / 2026, page 25 / 214. 15 / 195 acids from the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in SEQ ID NOs: 32 and 46.

[037] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region and a variable light chain region, wherein the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46. In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 25 and 46.In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 28 and 46. In certain embodiments, the variable heavy chain region and the variable light chain region, respectively, comprise the amino acid sequences presented in SEQ ID NOs: 32 and 46.

[038] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region and a variable light chain region, wherein the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in the SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and Petition 870260065483, dated 03 / 07 / 2026, p. 26 / 214 16 / 195 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46. In certain embodiments, the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in SEQ ID NOs: 25 and 46. In certain embodiments, the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in SEQ ID NOs: 28 and 46. In certain embodiments, the amino acid sequences of the variable heavy chain region and the variable light chain region, respectively, consist of the amino acid sequences presented in SEQ ID NOs: 32 and 46.

[039] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 58, and a light chain comprising the amino acid sequence of SEQ ID NO: 69.

[040] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 58, and a light chain comprising the amino acid sequence of SEQ ID NO: 69.

[041] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 61, and a light chain comprising the amino acid sequence of SEQ ID NO: 69.

[042] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61, and a light chain comprising the amino acid sequence of SEQ ID NO: 69. Petition 870260065483, dated 03 / 07 / 2026, p. 27 / 214 17 / 195

[043] In another aspect, the present disclosure provides an antibody or isolated antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain comprising the amino acid sequence of SEQ ID NO: 69.

[044] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 65, and a light chain comprising the amino acid sequence of SEQ ID NO: 69.

[045] In certain embodiments, the N-terminal glutamate (E) residue of an antibody heavy chain as described in this document is replaced by a pyroglutamate (pE) residue and / or the N-terminal glutamate (E) residue of an antibody light chain is replaced by a pyroglutamate (pE) residue.

[046] In certain embodiments, the antibody comprises a variable heavy chain region that has an amino acid sequence derived from a human IGHV3-23 germline sequence. In certain embodiments, the antibody comprises a variable light chain region that has an amino acid sequence derived from a human germline sequence selected from the group consisting of IGKV1-27, IGKV3-11, IGKV3-20, and IGKV3D-20.

[047] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody comprises a variable heavy chain region having an amino acid sequence derived from a germline sequence of human IGHV3-23, and a variable light chain region having an amino acid sequence derived from a human germline sequence selected from the group consisting of IGKV1-27, IGKV3-11, IGKV3-20 and IGKV3D-20. Petition 870260065483, dated 03 / 07 / 2026, p. 28 / 214 18 / 195

[048] In certain embodiments, the antibody comprises a constant chain region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In certain embodiments, the constant heavy chain region is IgG1. In certain embodiments, the amino acid sequence of IgG1 comprises an N297A mutation, numbered according to the EU numbering system. In certain embodiments, the antibody comprises a constant heavy chain region comprising the amino acid sequence SEQ ID NO: 72. In certain embodiments, the amino acid sequence of IgG1 comprises an N297Q mutation, numbered according to the EU numbering system. In certain embodiments, IgG1 is non-fucosylated IgG1. In certain embodiments, the constant heavy chain region is IgG4. In certain embodiments, the amino acid sequence of IgG4 comprises an S228P mutation, numbered according to the EU numbering system.In certain embodiments, the antibody comprises a constant heavy chain region comprising the amino acid sequence SEQ ID NO: 74.

[049] In certain embodiments, the antibody comprises a constant light chain region selected from the group consisting of human IgGK and ^λ. In certain embodiments, the constant light chain region is IgGK. In certain embodiments, the antibody comprises a constant light chain region comprising the amino acid sequence of SEQ ID NO: 76. In certain embodiments, the constant light chain region is ^λ.

[050] In another aspect, the present disclosure provides an antibody or isolated antibody that cross-competes for binding to human TIM-3 with an antibody as disclosed herein. In certain embodiments, the present disclosure provides an antibody or isolated antibody that cross-competes for binding to human TIM-3 with an antibody comprising the heavy and light chain variable region amino acid sequences presented in Petition 870260065483, dated 03 / 07 / 2026, p. 29 / 214 19 / 195 SEQ ID Nos: 55 and 56, respectively. In certain embodiments, this disclosure provides an antibody or isolated antibody that cross-competes for binding to human TIM-3 with an antibody comprising the variable region amino acid sequences of heavy and light chains presented in SEQ ID Nos: 25 and 46, respectively. In certain embodiments, this disclosure provides an antibody or isolated antibody that cross-competes for binding to human TIM-3 with an antibody comprising the variable region amino acid sequences of heavy and light chains presented in SEQ ID Nos: 28 and 46, respectively. In certain embodiments, this disclosure provides an antibody or isolated antibody that cross-competes for binding to human TIM-3 with an antibody comprising the variable region amino acid sequences of heavy and light chains presented in SEQ ID Nos: 32 and 46, respectively.

[051] In another aspect, the present disclosure provides an antibody or isolated antibody that binds to the same human TIM-3 epitope as an antibody disclosed in this document. In certain embodiments, the present disclosure provides an antibody or isolated antibody that binds to the same human TIM-3 epitope as an antibody comprising the heavy and light chain variable region amino acid sequences presented in SEQ ID NOs: 55 and 56, respectively. In certain embodiments, the present disclosure provides an antibody or isolated antibody that binds to the same human TIM-3 epitope as an antibody comprising the heavy and light chain variable region amino acid sequences presented in SEQ ID NOs: 25 and 46, respectively.In certain embodiments, the present disclosure provides an antibody or isolated antibody that binds to the same epitope of human TIM-3 as an antibody comprising the variable region amino acid sequences of heavy and light chains presented in SEQ IDs 28 and 46, respectively. In certain embodiments, the present disclosure provides an antibody or isolated antibody that binds to the same epitope of TIM. Petition 870260065483, dated 03 / 07 / 2026, p. 30 / 214 20 / 195 human as an antibody comprising the variable region amino acid sequences of heavy and light chains presented in SEQ ID NOs: 32 and 46, respectively.

[052] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody specifically binds to a variant TIM-3 protein that has the amino acid sequence SEQ ID NO: 101 with a lower affinity than to a wild-type TIM3 protein that has the amino acid sequence SEQ ID NO: 79.

[053] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In one embodiment, the antibody specifically binds to a variant TIM-3 protein having the amino acid sequence SEQ ID NO: 101 with a lower affinity than to a wild-type TIM-3 protein having the amino acid sequence SEQ ID NO: 79.

[054] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody does not specifically bind to a variant TIM-3 protein that has the amino acid sequence of SEQ ID NO: 101.

[055] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In one embodiment, the antibody does not specifically bind to a variant TIM-3 protein that has the amino acid sequence of SEQ ID NO: 101.

[056] In another aspect, the present disclosure provides an antibody or isolated antibody that binds specifically to human TIM-3, wherein the binding between the antibody and a variant TIM-3 protein that has the amino acid sequence of Petition 870260065483, dated 03 / 07 / 2026, p. 31 / 214 21 / 195 SEQ ID NO: 101 is substantially weakened in relation to the binding between the antibody and a wild-type TIM-3 protein that has the amino acid sequence of SEQ ID NO: 79.

[057] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In one embodiment, the binding between the antibody and a variant TIM-3 protein having the amino acid sequence SEQ ID NO: 101 is substantially weakened relative to the binding between the antibody and a wild-type TIM-3 protein having the amino acid sequence SEQ ID NO: 79.

[058] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to human TIM-3, wherein the antibody exhibits, compared with binding to a wild-type TIM-3 protein having the amino acid sequence SEQ ID NO: 79, reduced or absent binding to a variant TIM-3 protein having the amino acid sequence SEQ ID NO: 101.

[059] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In one embodiment, the antibody exhibits, compared with binding to a wild-type TIM-3 protein having the amino acid sequence SEQ ID NO: 79, reduced or absent binding to a variant TIM-3 protein having the amino acid sequence SEQ ID NO: 101.

[060] In another aspect, the present disclosure provides an antibody or isolated antibody that binds, for example, specifically binds, to a human TIM3 epitope. In certain embodiments, the antibody binds to residue 40 of SEQ ID NO: 79.

[061] In another aspect, the present disclosure provides an antibody or isolated antibody that binds specifically to the same human TIM-3 epitope that Petition 870260065483, dated 03 / 07 / 2026, p. 32 / 214 22 / 195 any antibody of the present invention. In certain embodiments, the antibody binds to residue 40 of SEQ ID NO: 79.

[062] In another aspect, the present disclosure provides an antibody or isolated antibody that binds, for example, specifically binds, to a human TIM3 epitope. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 93. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 94. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 95. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 96. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 97.In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 98. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 99. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 100.

[063] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 93. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 94. In certain embodiments, the antibody binds to an epitope Petition 870260065483, dated 03 / 07 / 2026, p. 33 / 214 23 / 195 located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 95. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 96. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 97. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 98. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 99. In certain embodiments, the antibody binds to an epitope located in a region of human TIM-3 consisting of the amino acid sequence SEQ ID NO: 100.

[064] In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 93, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 93 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 94, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 94 in the absence of the antibody, as determined by a hydrogen / deuterium assay. In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof that... Petition 870260065483, dated 03 / 07 / 2026, page 34 / 214 24 / 195 comprises the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 95, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 95 in the absence of the antibody, as determined by a hydrogen / deuterium assay. In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 96, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 96 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 97, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 97 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In another aspect, the present disclosure provides an antibody that, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 98, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 98 in the absence of the antibody, as determined by a hydrogen / deuterium assay. In some embodiments, the reduction in hydrogen / deuterium exchange is measured using hydrogen-deuterium exchange (HDX), for example, as described in the examples herein. Petition 870260065483, dated 03 / 07 / 2026, page 35 / 214 25 / 195

[065] In another aspect, the present disclosure provides an antibody or isolated antibody that specifically binds to the same human TIM-3 epitope as any antibody of the present invention. In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 93, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 93 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 94, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 94 in the absence of the antibody, as determined by a hydrogen / deuterium assay. In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 95, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 95 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 96, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 96 in the absence of the antibody, as determined by an assay of... Petition 870260065483, dated 03 / 07 / 2026, page 36 / 214 26 / 195 hydrogen / deuterium. In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 97, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 97 in the absence of the antibody, as determined by a hydrogen / deuterium assay.In certain embodiments, the antibody, when bound to a human TIM-3 protein or fragment thereof comprising the amino acid sequence of SEQ ID NO: 102, reduces hydrogen / deuterium exchange in a region consisting of the amino acid sequence presented in SEQ ID NO: 98, relative to hydrogen / deuterium exchange in the region consisting of the amino acid sequence presented in SEQ ID NO: 98 in the absence of the antibody, as determined by a hydrogen / deuterium assay. In some embodiments, the reduction in hydrogen / deuterium exchange is measured using hydrogen-deuterium exchange (HDX), for example, as described in the examples herein.

[066] In another aspect, the present disclosure provides an antibody or isolated antibody that binds, for example, specifically binds, to the same human TIM-3 epitope as any antibody of the present invention, wherein the epitope is determined by hydrogen-deuterium exchange (HDX), for example, as described in the examples, by Pepscan analysis, for example, as described in the examples, or by alanine scanning, for example, as described in the examples.

[067] In certain embodiments, the antibody comprises a constant region of human IgG heavy chain that is a variant of a wild-type human IgG heavy chain constant region, wherein the variant human IgG heavy chain constant region binds to a human Fc gamma receptor with lower affinity than the wild-type human IgG heavy chain constant region binds to the human Fc gamma receptor. In certain embodiments, the receptor of Petition 870260065483, dated 03 / 07 / 2026, p. 37 / 214 27 / 195 Human Fc gamma is selected from the group consisting of FcyRI, FcyRII, and FcyRIII. In certain embodiments, the variant human IgG heavy chain constant region is an IgG1 constant region comprising an N297A mutation.

[068] In certain embodiments, the antibody is a human antibody. In certain embodiments, the antibody is antagonistic to human TIM-3. In certain embodiments, the antibody inactivates, reduces, or inhibits an activity of human TIM-3. In certain embodiments, the antibody inhibits the binding of human TIM-3 to phosphatidylserine. In certain embodiments, the antibody induces the production of IFNγ by peripheral blood mononuclear cells (PBMCs) stimulated with staphylococcal enterotoxin A (SEA). In certain embodiments, the antibody induces the production of IFNγ or TNFα by tumor-infiltrating lymphocytes (TILs) stimulated with anti-CD3 and anti-CD28 antibodies.

[069] In certain embodiments, the antibody is internalised after binding to cells that express human TIM-3.

[070] In another aspect, the present disclosure provides an antibody or isolated antibody as disclosed herein conjugated to a cytotoxic agent.

[071] In another aspect, the present disclosure provides an antibody or isolated antibody as disclosed herein conjugated to a cytostatic agent.

[072] In another aspect, the present disclosure provides an antibody or isolated antibody as disclosed herein conjugated to a toxin.

[073] In another aspect, the present disclosure provides an antibody or isolated antibody as disclosed herein conjugated to a radionuclide.

[074] In another aspect, the present disclosure provides an antibody or isolated antibody as disclosed herein conjugated to a detectable marker. Petition 870260065483, dated 03 / 07 / 2026, page 38 / 214 28 / 195

[075] In another aspect, the present disclosure provides a pharmaceutical composition comprising an antibody as disclosed herein and a pharmaceutically acceptable carrier or excipient.

[076] In another aspect, the present disclosure provides a polynucleotide or isolated polynucleotide encoding a heavy and / or light chain of an antibody as disclosed herein. In another aspect, the present disclosure provides a vector comprising the polynucleotide. In another aspect, the present disclosure provides a recombinant host cell comprising the polynucleotide. In another aspect, the present disclosure provides a recombinant host cell comprising the vector. In another aspect, the present disclosure provides a method for producing an antibody as disclosed herein, wherein the method comprises culturing the host cell so that the polynucleotide is expressed and the antibody is produced. In one embodiment, the method is an in vitro method.

[077] In one embodiment, the present invention relates to an antibody of the invention, or a pharmaceutical composition of the invention, or a polynucleotide of the invention, or a vector of the invention, or a recombinant host cell of the invention for use as a medicament.

[078] In one embodiment, the present invention relates to an antibody of the invention, or a pharmaceutical composition of the invention, or a polynucleotide of the invention, or a vector of the invention, or a recombinant host cell of the invention for use as a diagnostic.

[079] In another aspect, the present disclosure provides a method for increasing T cell activation in response to an antigen in an individual, wherein the method comprises administering to the individual an effective amount of an antibody or pharmaceutical composition as disclosed herein. In another aspect, the present disclosure provides a method for treating cancer in an individual, Petition 870260065483, dated 03 / 07 / 2026, p. 39 / 214 29 / 195 wherein the method comprises administering to the individual an effective amount of an antibody or pharmaceutical composition as disclosed in this document. In certain embodiments of the foregoing methods, the antibody or pharmaceutical composition is administered subcutaneously. In certain embodiments of the foregoing methods, the antibody or pharmaceutical composition is administered intravenously. In certain embodiments of the foregoing methods, the antibody or pharmaceutical composition is administered intratumorally. In certain embodiments of the foregoing methods, the antibody or pharmaceutical composition is delivered to a tumor-draining lymph node. In certain embodiments of the foregoing methods, the antibody or pharmaceutical composition is administered intra-arterially.

[080] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for enhancing T cell activation in response to an antigen.

[081] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for increasing T cell activation in response to an antigen in an individual.

[082] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for increasing T cell activation in response to an antigen in an individual comprising administering to the individual an effective amount of an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the invention.

[083] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for the treatment of cancer. Petition 870260065483, dated 03 / 07 / 2026, page 40 / 214 30 / 195

[084] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for treating cancer in an individual.

[085] In one aspect, the present invention relates to an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention for use in a method for treating cancer in an individual comprising administering to the individual an effective amount of an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the invention.

[086] In one embodiment of an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition for use in the present invention, the antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition are administered subcutaneously or intravenously. In another embodiment of an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition for use in the present invention, the antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition are administered intratumorally or intra-arterially.

[087] In certain embodiments, the foregoing methods additionally comprise administering an additional therapeutic agent to the individual. Therefore, in an embodiment of an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition for use in a method of the present invention, the method additionally comprises administering an additional therapeutic agent to the individual.

[088] In one aspect, the present invention relates to (a) an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention and (b) an additional therapeutic agent for use as a medicament. Petition 870260065483, dated 03 / 07 / 2026, p. 41 / 214 31 / 195

[089] In one aspect, the present invention relates to (a) an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention and (b) an additional therapeutic agent for use in a method for treating cancer.

[090] In one aspect, the present invention relates to a pharmaceutical composition, kit or kit of parts comprising (a) an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention and (b) an additional therapeutic agent.

[091] In certain modalities, the additional therapeutic agent is a chemotherapeutic product. In certain modalities, the additional therapeutic agent is a radiotherapeutic product.

[092] In certain embodiments, the additional therapeutic agent is a checkpoint targeting agent. In certain embodiments, the checkpoint targeting agent is selected from the group consisting of an anti-PD1 antagonist antibody, an anti-PD-L1 antagonist antibody, an anti-PD-L2 antagonist antibody, an anti-CTLA-4 antagonist antibody, an anti-TIM-3 antagonist antibody, an anti-LAG-3 antagonist antibody, an anti-CEACAM1 antagonist antibody, an anti-CD137 agonist antibody, an anti-TIGIT antagonist antibody, an anti-VISTA antagonist antibody, an anti-GITR antagonist antibody, and an anti-OX40 agonist antibody. In certain embodiments, the additional therapeutic agent is an anti-PD-1 antibody. In certain embodiments, the anti-PD-1 antibody is pembrolizumab. In certain modalities, the anti-PD-1 antibody is nivolumab.

[093] In certain embodiments, the additional therapeutic agent is an indoleamine-2,3-dioxygenase (IDO) inhibitor. In certain embodiments, the inhibitor is selected from the group consisting of epacadostat, F001287, indoximod, and NLG919. In certain embodiments, the inhibitor is epacadostat. In certain embodiments, the inhibitor is F001287. In certain embodiments, the inhibitor is indoximod. In certain embodiments, the inhibitor is Petition 870260065483, dated 03 / 07 / 2026, page 42 / 214 32 / 195 NLG919.

[094] In certain embodiments, the additional therapeutic agent is a vaccine. In certain embodiments, the vaccine comprises a heat shock protein-peptide complex (HSPPC) comprising a heat shock protein complexed with an antigenic peptide. In certain embodiments, the heat shock protein is hsc70 and is complexed with a tumor-associated antigenic peptide. In certain embodiments, the heat shock protein is gp96 protein and is complexed with a tumor-associated antigenic peptide, wherein the HSPPC is derived from a tumor obtained from an individual. In certain embodiments, the additional therapeutic agent comprises a TCR. In certain embodiments, the additional therapeutic agent is a soluble TCR. In certain embodiments, the additional therapeutic agent is a cell expressing a TCR. In certain embodiments, the additional therapeutic agent is a cell expressing a chimeric antigen receptor.In certain embodiments, the additional therapeutic agent is an antibody that binds specifically to a peptide-MHC complex. In certain embodiments, the additional therapeutic agent is an adjuvant. In one aspect, the present invention relates to (a) an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention and (b) a vaccine for use as a medicament, for example, for use in a method for the treatment of cancer, optionally, wherein the vaccine comprises a heat shock protein and peptide complex (HSPPC) comprising a heat shock protein complexed with an antigenic peptide.In one aspect, the present invention relates to a pharmaceutical composition, kit or kit of parts comprising (a) an antibody, polynucleotide, vector, recombinant host cell and / or pharmaceutical composition of the present invention and (b) a vaccine, optionally, wherein the vaccine comprises a heat shock protein and peptide complex (HSPPC) comprising a heat shock protein complexed with an antigenic peptide. Petition 870260065483, dated 03 / 07 / 2026, page 43 / 214 33 / 195 5. BRIEF DESCRIPTION OF THE DRAWINGS

[095] Figure 1 is a set of histograms showing the binding of anti-TIM-3 antibodies pab2085 (IgG1) and pab2088 (IgG1) or an isotype control antibody to wild-type murine 1624-5 cells or 1624-5 cells genetically modified to express human TIM-3, as measured by flow cytometry.

[096] Figures 2A and 2B are a pair of graphs showing the binding of anti-TIM-3 antibodies pab2085 (IgG1) (Figure 2A) and pab2088 (IgG1) (Figure 2B) to recombinant human TIM-1 His (rhTIM-1 His), recombinant human TIM-4 His (rhTIM-4 His), recombinant human TIM-3 His (rhTIM-3 His), recombinant human TIM-3 Fc (rhTIM-3 Fc), and recombinant cynomolgus TIM-3 Fc (rcmTIM-3 Fc), as measured by Luminex assay. Median fluorescence intensity (MFI) values ​​are plotted against antibody concentrations.

[097] Figures 3A, 3B, 3C and 3D are a set of histograms showing the binding of anti-TIM-3 antibodies or an isotype control antibody to murine 1624-5 cells genetically modified to express human TIM-3 (Figures 3A and 3B) or cynomolgus TIM-3 (Figures 3C and 3D), as measured by flow cytometry. The anti-TIM-3 antibodies tested in this study include pab2173, pab2174, pab2175, pab2176, pab2177, pab2178, pab2179, pab2180, pab2181, pab2182, pab2183, pab2184, pab2185, pab2186, pab2187, pab2188, pab2189, pab2190, pab2191, and pab2192, all of which contain an Fc region of IgG1.

[098] Figure 4 is a graph showing the binding of anti-TIM-3 antibody pab2085, optimized light chain variants (pab2184, pab2186, pab2187, pab2188, pab2189, pab2190, pab2191, and pab2192), or an isotype control antibody to primary human CD8+ T cells activated by anti-CD3 and anti-CD28 antibodies, as measured by flow cytometry. Optimized light chain variants contain an IgG1 variant Fc region. MFI values ​​are plotted against a range of antibody concentrations tested. Petition 870260065483, dated 03 / 07 / 2026, p. 44 / 214 34 / 195

[099] Figure 5 is a graph showing the binding of anti-TIM-3 pab2188 antibody (IgG1 variant) or an isotype control antibody to primary cynomolgus CD11b+ myeloid cells, as measured by flow cytometry. MFI values ​​are plotted against antibody concentrations.

[0100] Figures 6A and 6B are graphs showing the percentage of binding between irradiated murine WR19L lymphoma cells expressing phosphatidylserine and Fc of recombinant human TIM-3 (Figure 6A) or Fc of recombinant cynomolgus TIM-3 (Figure 6B) in the presence of a dose titration of an anti-TIM-3 antibody or an IgG1 isotype control antibody. The anti-TIM-3 antibodies tested in this study are pab2085 (IgG1) and pab2188 (IgG1 variant).

[0101] Figure 7 is a bar graph showing IFNγ production induced by anti-TIM-3 antibodies or an IgG1 isotype control antibody in combination with the anti-PD-1 pembrolizumab antibody in human peripheral blood mononuclear cells (PBMCs) after stimulation with Staphylococcus Enterotoxin A (SEA). The anti-TIM-3 antibodies tested in this study include optimized light chain variants pab2175 (IgG1), pab2176 (IgG1), pab2180 (IgG1), pab2182 (IgG1), pab2183 (IgG1 variant), pab2184 (IgG1 variant), pab2186 (IgG1 variant), pab2187 (IgG1 variant), pab2188 (IgG1 variant), pab2189 (IgG1 variant), pab2190 (IgG1 variant), pab2191 (IgG1 variant), and pab2192 (IgG1 variant).

[0102] Figures 8A, 8B, 8C, 8D, 8E, and 8F are a set of bar graphs showing IFNγ production induced by the anti-TIM-3 pab2188w antibody (IgG1 N297A) or an IgG1 isotype N297A control antibody, alone or in combination with the anti-PD-1 pembrolizumab antibody, in human PBMCs from six different donors after stimulation with SEA. The protocol used in the study represented in Figures 8A to 8F was modified from the protocol used in the study represented in Figure 7. Petition 870260065483, dated 03 / 07 / 2026, page 45 / 214 35 / 195

[0103] Figures 9A, 9B, 9C, 9D, 9E, and 9F are graphs or histograms showing the binding of anti-TIM-3 antibodies to cells expressing TIM-3. In Figures 9A, 9B, 9E, and 9F, MFI values ​​are plotted against a range of antibody concentrations tested. Figures 9C and 9D are a set of histograms showing the binding of anti-TIM-3 antibodies to cells expressing TIM-3. Anti-TIM-3 antibodies tested include pab2188w (IgG1 N297A), AM-1 (IgG1 N297A), AM-2 (IgG1 N297A), AM-3 (IgG1 N297A), AM-4 (IgG1 N297A), AM-5 (IgG1 N297A), AM-6 (IgG1 N297A), AM-7 (IgG1 N297A), AM-8 (IgG1 N297A) and AM-9 (IgG1 N297A).The cells tested were Jurkat cells ectopically expressing human TIM-3 (Figure 9A), Kasumi-3, a human acute myeloid leukemia cell line endogenously expressing TIM-3 (Figure 9B), human CD8+ T cells stimulated with Staphylococcal Enterotoxin A (SEA) (Figure 9C), cynomolgus CD8+ T cells stimulated with SEA (Figure 9D), and primary human (Figure 9E) and cynomolgus (Figure 9F) CD14+ myeloid cells.

[0104] Figures 10A, 10B, 10C and 10D are graphs showing the binding of anti-TIM-3 antibodies or an N297A IgG1 isotype control antibody to recombinant human TIM-3 His (rhTIM-3 His), recombinant cynomolgus TIM-3 Fc (rcmTIM-3 Fc), recombinant mouse TIM-3 Fc (rmTIM-3 Fc), recombinant human TIM1 His (rhTIM-1 His), recombinant human TIM-4 His (rhTIM-4 His), recombinant human OX40 His (rhOX40 His), recombinant human GITR Fc (rhGITR Fc), recombinant human DR3 Fc (rhDR3 Fc) and recombinant human CD137 Fc (rhCD137 Fc), measured by a Luminex assay. MFI values ​​are plotted against antibody concentrations. The anti-TIM-3 antibodies tested in this study include pab2188w (IgG1 N297A) (Figure 10B), AM-2 (IgG1 N297A) (Figure 10C), and AM-6 (IgG1 N297A) (Figure 10D).

[0105] Figures 11A and 11B are graphs showing the percentage of Fc binding of recombinant human TIM-3 (Figure 11A) or Fc of cynomolgus TIM-3. Petition 870260065483, dated 03 / 07 / 2026, p. 46 / 214 36 / 195 recombinant (Figure 11B) to WR19L cells expressing phosphatidylserine in the presence of a dose titration of anti-TIM-3 antibody or an N297A isotype control antibody of IgG1. The anti-TIM-3 antibodies tested in this study include pab2188w (IgG1 N297A), AM-2 (IgG1 N297A), and AM-6 (IgG1 N297A).

[0106] Figures 12A and 12B are bar graphs showing IFNγ production induced by anti-TIM-3 antibodies or an IgG1 isotype N297A control antibody, alone or in combination with the anti-PD-1 pembrolizumab antibody, in human PBMCs from two different donors after SEA stimulation. The anti-TIM-3 antibodies tested include pab2188w (IgG1 N297A), AM-1 (IgG1 N297A), AM-2 (IgG1 N297A), AM-3 (IgG1 N297A), AM-4 (IgG1 N297A), AM-5 (IgG1 N297A), AM-6 (IgG1 N297A), AM-7 (IgG1 N297A), and AM-8 (IgG1 N297A).

[0107] Figures 13A, 13B, 13C, 13D, 13E, and 13F are graphs showing the production of IFNγ or TNFα by primary tumor-infiltrating lymphocytes (TILs) induced by anti-TIM-3 antibodies or an N297A isotype control antibody of IgG1, alone or in combination with the anti-PD-1 antibody pembrolizumab. The TILs were isolated from non-small cell lung cancer (NSCLC) tumors (Figures 13A and 13B), gallbladder adenocarcinoma (Figures 13C and 13D), or breast cancer (Figures 13E and 13F) and activated with anti-CD3 / CD28 microspheres. The anti-TIM-3 antibodies tested in this study include pab2188w (IgG1 N297A), AM-2 (IgG1 N297A), and AM-6 (IgG1 N297A).

[0108] Figures 14A, 14B, and 14C are graphs showing the percentage of cell survival, relative to an untreated control group, after incubation with an anti-TIM-3 antibody or an N297A isotype control antibody of IgG1. Figures 14A and 14B show treatment with the indicated antibody in combination with an antibody-secondary drug conjugate αHFc-NC-DM1. The cells tested were Jurkat cells genetically modified for superex Petition 870260065483, dated 03 / 07 / 2026, page 47 / 214 37 / 195 pressar TIM-3 (Figure 14A) or Kasumi-3 cells, an acute myeloid leukemia cell line that endogenously expresses TIM-3 (Figure 14B). Figure 14C shows treatment with the indicated antibody as a conjugate with monomethyl auristatin E (MMAE). The anti-TIM-3 antibodies tested in this study include pab2188w (Ig297G1 NA), AM-2 (IgG1 N297A), AM-6 (IgG1 N297A), and the reference antibodies Hum11 (IgG4 S228P) and pab1944w (IgG1 N297A).

[0109] Figure 15 is a series of graphs showing the internalization of TIM-3 in Jurkat cells expressing a HaloTag-TIM-3 fusion protein when incubated with 10 μg / ml of anti-TIM-3 AM-2 antibody or an isotype control antibody, as determined by live-cell confocal fluorescence microscopy, at various time points (i.e., 0 to 3.5 hours). Black dots indicate the average fluorescence level observed for each condition at a given time point. 6. DETAILED DESCRIPTION

[0110] This disclosure provides antibodies that specifically bind to TIM-3 (e.g., human TIM-3) and antagonize TIM-3 function, e.g., TIM-3-mediated immune suppression. Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for producing these antibodies, and methods of treating an individual using these antibodies are also provided. The antibodies disclosed herein are particularly useful for increasing T cell activation in response to an antigen (e.g., a tumor antigen or an infectious disease antigen) and therefore for treating cancer in an individual or treating or preventing an infectious disease in an individual. All instances of “isolated antibodies” described herein are further contemplated as antibodies that may be, but need not be, isolated. All instances of Petition 870260065483, dated 03 / 07 / 2026, page 48 / 214 38 / 195 “isolated polynucleotides” described in this document are additionally considered as polynucleotides that may be, but do not need to be, isolated. All cases of “antibodies” described in this document are additionally considered as antibodies that may be, but do not need to be, isolated. All cases of “polynucleotides” described in this document are additionally considered as polynucleotides that may be, but do not need to be, isolated. 6.1 DEFINITIONS

[0111] As used in this document, the terms “about” and “approximately”, when used to modify a numerical value or a numerical range, indicate that deviations of 5% to 10% above (e.g., up to 5% to 10% above) and 5% to 10% below (e.g., up to 5% to 10% below) the value or range remain within the intended meaning of the value or range mentioned.

[0112] As used in this document, the term “TIM-3” refers to a T-cell mucin and immunoglobulin domain 3 (also known as T-cell mucin and immunoglobulin domain-containing protein 3 or Hepatitis A virus cell receptor 2 (HAVCR2)) which in humans is encoded by the HAVCR2 gene. Swiss-Prot accession number Q8TDQ0-1 provides an exemplary human TIM-3 amino acid sequence. The immature human TIM-3 amino acid sequence is provided as SEQ ID NO: 78. The mature human TIM-3 amino acid sequence is provided as SEQ ID NO: 79. As used in this document, the term “human TIM-3” refers to TIM-3 comprising the amino acid sequence of SEQ ID NO: 79.

[0113] As used in this document, the terms “antibody” and “antibodies” include full-length antibodies, antigen-binding fragments of full-length antibodies, and molecules comprising CDRs, VH regions, or VL regions of antibody. Examples of antibodies include Petition 870260065483, dated 03 / 07 / 2026, page 49 / 214 39 / 195 monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, a monovalent antibody heavy chain, an antibody light chain dimer, an antibody heavy chain dimer, an antibody light chain and antibody heavy chain pair, intracorporeal antibodies, heteroconjugated antibodies, antibody-drug conjugates, single-domain antibodies, monovalent antibodies, single-chain antibodies or single-chain Fvs (scFv), camelized antibodies, affybodies, Fab fragments, F(ab')2 fragments, disulfide-linked Fvs (sdFv), anti-idiotypic antibodies (anti-Id) (including, for example,Anti-anti-Id antibodies) and antigen-binding fragments of any of the above. In certain embodiments, the antibodies described herein refer to polyclonal antibody populations. The antibodies may be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgGa, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In certain embodiments, the antibodies described herein are IgG antibodies or a class (e.g., human IgG1 or IgG4 or a subclass thereof). In one specific embodiment, the antibody is a humanized monoclonal antibody. In another specific embodiment, the antibody is a human monoclonal antibody.

[0114] As used in this document, the terms “VH region” and “VL region” refer to single antibody heavy and light chain variable regions, respectively, comprising FR (Framework Regions) 1, 2, 3, and 4 and CDR (Complementarity Determinant Regions) 1, 2, and 3 (see Rabat et al., (1991) Sequences of Proteins of Immunological Interest (NIH Publication No. 913242, Bethesda), which is incorporated herein by reference). Petition 870260065483, dated 03 / 07 / 2026, p. 50 / 214 40 / 195 in its entirety).

[0115] As used in this document, the term CDR or complementarity-determining region means non-contiguous antigen-binding sites found within the variable region of both heavy and light chain polypeptides. These particular regions were described by Rabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Rabat et al., Sequences of proteins of immunological interest. (1991), by Chothia et al., J. Mol. Biol. 196:901-917 (1987), and by MacCallum et al., J. Mol. Biol. 262:732-745 (1996), all of which are incorporated herein by reference in their entirety, wherein the definitions include overlapping or subsets of amino acid residues compared to one another. In certain embodiments, the term “CDR” is a CDR as defined by MacCallum et al., J. Mol. Biol. 262:732-745 (1996) and Martin A. “Protein Sequencing and Variable Domain Structure Analysis of Antibodies” in Antibody Engineering, Rontermann and Dtibel, eds.Chapter 31, pages 422-439, Springer-Verlag, Berlin (2001). In certain embodiments, the term “CDR” is a CDR as defined by Rabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Rabat et al., Sequences of protein of immunological interest (1991). In certain embodiments, the heavy chain CDRs and the light chain CDRs of an antibody are defined using different conventions. For example, in certain embodiments, heavy chain CDRs are defined according to MacCallum (above), and light chain CDRs are defined according to Rabat (above). CDRH1, CDRH2, and CDRH3 denote the heavy chain CDRs, and CDRL1, CDRL2, and CDRL3 denote the light chain CDRs.

[0116] As used in this document, the term “framework amino acid residues (FR)” refers to those amino acids in the framework region of an immunoglobulin chain. The term “framework region” or “FR region,” as used in this document, includes amino acid residues that are part of the variable region but are not part of the CDRs (e.g., with the use of Petition 870260065483, dated 03 / 07 / 2026, page 51 / 214 41 / 195 definition of Rabat or MacCallum of CDRs).

[0117] As used in this document, the terms “variable region” and “variable domain” are used interchangeably and are common in the art. The variable region typically refers to a portion of an antibody, generally a portion of a light or heavy chain, typically about 110 to 120 amino-terminal amino acids or 110 to 125 amino acids in the mature heavy chain and about 90 to 115 amino-terminal amino acids in the mature light chain, which differs extensively in sequence between antibodies and is used in the binding and specificity of a particular antibody to its particular antigen. Variability in sequence is concentrated in those regions called complementarity-determining regions (CDRs) while the most highly conserved regions in the variable domain are called frame regions (FRs).Without wishing to adhere to any particular mechanism or theory, it is believed that CDRs of light chains and chains are primarily responsible for antibody-antigen interaction and specificity. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and human scaffold regions (FRs). In particular embodiments, the variable region is a primate variable region (e.g., non-human primate). In certain embodiments, the variable region comprises rodent or murine CDRs and primate scaffold regions (e.g., non-human primate) (FRs).

[0118] The terms “VL” and “VL domain” are used interchangeably to refer to the variable light chain region of an antibody.

[0119] The terms “VH” and “VH domain” are used interchangeably to refer to the variable heavy chain region of an antibody.

[0120] As used in this document, the terms “constant region” and “constant domain” are interchangeable and are common in the art. The constant region is a portion of antibody, for example, a carboxyl-terminal portion of Petition 870260065483, dated 03 / 07 / 2026, page 52 / 214 42 / 195 a light and / or heavy chain that is not directly involved in the binding of an antibody to an antigen, but which may exhibit various effector functions, such as interaction with an Fc receptor (e.g., an Fc gamma receptor). The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence compared to a variable domain of immunoglobulin.

[0121] As used in this document, the term “heavy chain”, when used in reference to an antibody, may refer to any distinct type, for example, alpha (α), delta (δ), epsilon (ε), gamma (γ) and mu (μ), based on the amino acid sequence of the constant domain, which generates the IgA, IgD, IgE, IgG and IgM classes of antibodies, respectively, including the IgG subclasses, for example, IgG1, IgG2, IgG3 and IgG4.

[0122] As used in this document, the term “light chain,” when used in reference to an antibody, may refer to any distinct type, for example, kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domains. Light chain amino acid sequences are well known in the art. In specific embodiments, the light chain is a human light chain.

[0123] As used in this document, the term “EU numbering system” refers to the EU numbering convention for the constant regions of an antibody, as described in Edelman, GM et al., Proc. Natl. Acad. USA, 63, 7885 (1969) and Kabat et al, Sequences of Proteins of Immunological Interest, US Dept. Health and Human Services, 5th edition, 1991, each of which is incorporated herein by reference in its entirety.

[0124] “Binding affinity” generally refers to the intensity of the total sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, “binding affinity” refers to intrinsic binding affinity which reflects Petition 870260065483, dated 03 / 07 / 2026, p. 53 / 214 43 / 195 a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured and / or expressed in various forms known in the art, including, but not limited to, equilibrium dissociation constant (Kd) and equilibrium association constant (Ka). Kd is calculated from the quotient of koff / kon, while Ka is calculated from the quotient of kon / koff. kon refers to the association rate constant of, for example, an antibody to an antigen, and koff refers to the dissociation rate constant of, for example, an antibody to an antigen. kon and koff can be determined by techniques known to the common skill in the art, such as BIAcore® or KinExA. As used in this document, a “lower affinity” refers to a higher Kd.

[0125] As used in this document, the terms “specifically binds,” “specifically recognizes,” “immunospecifically binds,” and “immunospecifically recognizes” are analogous terms in the context of antibodies and refer to molecules that bind to an antigen (e.g., epitope or immune complex) as such binding is understood by one skilled in the art. For example, a molecule that specifically binds to an antigen may bind to other peptides or polypeptides, generally with lower affinity as determined, for example, by immunoassays, BIAcore®, KinExA 3000 instrument (Sapidyne Instruments, Boise, ID), or other assays known in the art. In a specific embodiment, molecules that specifically bind to an antigen bind to the antigen with a Ka that is at least 2 logs (e.g., factors of 10), 2.5 logs, 3 logs, 4 logs, or more than the Ka when the molecules bind non-specifically to another antigen.

[0126] In another specific form, the molecules that bind specifically Petition 870260065483, dated 03 / 07 / 2026, p. 54 / 214 44 / 195 molecules that bind specifically to an antigen do not cross-react with other proteins under similar binding conditions. In another specific embodiment, molecules that bind specifically to TIM-3 do not cross-react with other non-TIM3 proteins. In a specific embodiment, an antibody is provided herein that binds to TIM-3 (e.g., human TIM-3) with higher affinity than to another unrelated antigen. In certain embodiments, an antibody is provided herein that binds to TIM-3 (e.g., human TIM-3) with an affinity of 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or higher than to another unrelated antigen, as measured, for example, by a radioimmunoassay, surface plasmon resonance or kinetic exclusion assay.In one specific embodiment, the extent of binding of an anti-TIM-3 antibody described herein to an unrelated non-TIM-3 protein is less than 10%, 15%, or 20% of the antibody binding to the TIM-3 protein as measured, for example, by a radioimmunoassay.

[0127] As used in this document, the term “afucosylation” or “afucosylated” in the context of an Fc refers to a substantial lack of a fucose covalently linked, directly or indirectly, to residue 297 of the Fc region of human IgG1, numbered according to the EU numbering system, or the corresponding residue in non-IgG1 or non-human IgG1 immunoglobulins. Thus, in a composition comprising a plurality of afucosylated antibodies, at least 70% of the antibodies will not be fucosylated, directly or indirectly (e.g., via intervening sugars), at residue 297 of the Fc region of the antibodies and, in some embodiments, at least 80%, 85%, 90%, 95% or 99% will not be fucosylated, directly or indirectly, at residue 297 of the Fc region.

[0128] As used in this document, an “epitope” is a term in the art and refers to a localized region of an antigen with which an antibody Petition 870260065483, dated 03 / 07 / 2026, page 55 / 21445 / 195 can bind specifically. An epitope can be, for example, adjacent amino acids of a polypeptide (linear or contiguous epitope) or an epitope can, for example, be adjacent to two or more non-contiguous regions of a polypeptide or polypeptides (conformational, non-linear, discontinuous or non-contiguous epitope). In certain embodiments, the epitope to which an antibody binds can be determined, for example, by NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., electroscatter mass spectrometry coupled with liquid chromatography), array-based oligopeptide screening assays (e.g., restricting peptides with the use of CLIPS (Chemical Linking of Peptides on Scaffolds) to map discontinuous or conformational epitopes) and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping).For X-ray crystallography, crystallization can be performed using any of the methods known in the art (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; Chayen NE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303; each of which is incorporated herein by reference in its entirety). Antibody:antigen crystals can be studied using well-known X-ray diffraction techniques and can be refined using computer software such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985) volumes 114 and 115, eds Wyckoff HW et al.,; document no. US 2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A:361 to 423, ed Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323), each of which is incorporated herein by reference in its entirety. Mutagenesis mapping studies can be performed using any of these. Petition 870260065483, dated 03 / 07 / 2026, p. 56 / 214 46 / 195 methods known to those skilled in the art. See, for example, Champe M et al., (1995) J Biol Chem 270: 1388-1394 and Cunningham BC & Wells JA (1989) Science 244: 1081-1085, each of which is incorporated herein by reference in its entirety, for a description of mutagenesis techniques, including alanine scavenging mutagenesis techniques. CLIPS (Chemical Linking of Peptides in Scaffolds) is a technology for presenting one or more peptides in a structurally restricted configuration to behave as functional mimetics of complex protein domains. See, for example, U.S. Publications Nos. 2008 / 0139407 A1 and 2007 / 099240 A1 and U.S. Patent No. 7,972,993, each of which is incorporated herein by reference in its entirety. In one specific embodiment, the epitope of an antibody is determined using alanine-screening mutagenesis studies.In one specific embodiment, an antibody epitope is determined using hydrogen / deuterium exchange coupled with mass spectrometry. In another specific embodiment, an antibody epitope is determined using Pepscan Therapeutics' CLIPS Epitope Mapping Technology.

[0129] As used in this document, the term “an epitope located within a human TIM-3 region” consisting of a particular amino acid sequence or set of amino acid residues refers to an epitope comprising one or more amino acid residues from the specified region, wherein the specified region includes the first specified amino acid residue and the last specified amino acid residue of the human TIM-3 region. In certain embodiments, the epitope comprises each of the amino acid residues located within the specified region. In certain embodiments, one or more additional human TIM-3 amino acid residues outside the specified region bind to an antibody together with an epitope located within the specified region. Petition 870260065483, dated 03 / 07 / 2026, page 57 / 214 47 / 195

[0130] As used in this document, the terms “T cell receptor” and “TCR” are used interchangeably and refer to full-length heterodimeric αβ or γδ TCRs, antigen-binding fragments of full-length TCRs, and molecules comprising TCR CDRs or variable regions. Examples of TCRs include, but are not limited to, full-length TCRs, antigen-binding fragments of full-length TCRs, soluble TCRs lacking transmembrane and cytoplasmic regions, single-chain TCRs containing variable TCR regions bound by a flexible linker, genetically modified disulfide-linked TCR chains, monospecific TCRs, multispecific TCRs (including bispecific TCRs), TCR fusions, human TCRs, humanized TCRs, chimeric TCRs, recombinantly produced TCRs, and synthetic TCRs.The term encompasses wild-type TCRs and genetically modified TCRs (e.g., a chimeric TCR comprising a chimeric TCR chain that includes a first portion of a TCR from a first species and a second portion of a TCR from a second species).

[0131] As used in this document, the terms “major histocompatibility complex” and “MHC” are used interchangeably and refer to an MHC class I molecule and / or an MHC class II molecule.

[0132] As used in this document, the term “peptide-MHC complex” refers to an MHC molecule (MHC class I or MHC class II) with a peptide bound in the peptide-binding pocket recognized in the MHC technique.

[0133] As used in this document, the terms “treat,” “treating,” and “treatment” refer to therapeutic or preventive measures described herein. “Treatment” methods employ the administration of an antibody to an individual who has a disease or disorder or is predisposed to having such a disease or disorder, in order to prevent, cure, delay, reduce the severity of, or alleviate Petition 870260065483, dated 03 / 07 / 2026, p. 58 / 214 48 / 195 one or more symptoms of the disease or disorder or recurrent disease or disorder or in order to prolong an individual's survival beyond what would be expected in the absence of such treatment.

[0134] As used in this document, the term “effective amount” in the context of administering a therapy to an individual refers to the amount of a therapy that achieves a desired prophylactic or therapeutic effect.

[0135] As used in this document, the term “individual” includes any human being or non-human animal. In one embodiment, the individual is a human being or a non-human mammal. In another embodiment, the individual is a human being.

[0136] Determining the percentage of identity between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be performed using a mathematical algorithm. A specific non-limiting example of a mathematical algorithm used for comparing two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, each of which is incorporated herein by reference in its entirety. This algorithm is incorporated in the NBLAST and XBLAST programs of Altschul SF et al., (1990) J. Mol Biol 215: 403, which is incorporated herein by reference in its entirety.Nucleotide BLAST searches can be performed with the NBLAST nucleotide program parameters defined, for example, for score = 100, word length = 12 to obtain nucleotide sequences homologous to nucleic acid molecules described in this document. Protein BLAST searches can be performed with the XBLAST program parameters defined, for example, for score = 50, word length = 3 to obtain amino acid sequences homologous to a protein molecule described in this document. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul SF et al. Petition 870260065483, dated 03 / 07 / 2026, page 59 / 214 49 / 195 et al., (1997) Nucleic Acids Res 25: 3389-3402, which is incorporated herein by reference in its entirety. Alternatively, PSI BLAST can be used to perform a repeated search that detects distant relationships between molecules (Id.). After using the BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another non-limiting example of a specific mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is incorporated herein by reference in its entirety. This algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package.After using the ALIGN program to compare amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0137] The percentage of identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percentage identity, typically only exact correlations are counted.

[0138] As used in this document, the term “internalization” or “internalized” refers to the uptake of an antibody into an intracellular compartment of a cell after the antibody binds to an antigen expressed on the cell surface. 6.2 Anti-TIM-3 Antibodies

[0139] In one aspect, the present disclosure provides antibodies that bind specifically to TIM-3 (e.g., human TIM-3) and antagonize the function of Petition 870260065483, dated 03 / 07 / 2026, page 60 / 214 50 / 195 TIM-3. The amino acid sequences of exemplary antibodies are presented in Tables 1 to 4 in this document. TABLE 1. AMINO ACID SEQUENCES OF EXEMPLARY ANTI-TIM3 ANTIBODIES. SEQ ID NO: Description* Amino acid sequence 1 BADD456-2919 CDRH1 SSYAMS 2 BADD456-2919 CDRH2 WVSAISGSGGSTY 3 BADD456-2919 CDRH3 AKGGDYGGNYFD 4 AM-1 CDRH1 KAGQSS 5 AM-2 CDRH1 RQNAWS 6 AM-3 CDRH1 MSGQTS 7 AM-4 CDRH1 GAGQSS 8 AM-5 CDRH1 SAQQAS 9 AM-6 CDRH1 ARNAWS 10 AM-7 CDRH1 RSQQAS 11 AM-8 CDRH1 TTQQAS 12 AM-9 CDRH1 GGQQAS 13 BADD197-1181 CDRL1 RASQSVSSSYLA 14 BADD412-2513 CDRL1 RASQSVSSYLA 15 BADD456-2928 CDRL1 RASQGISNYLA 16 BADD466-3169 CDRL1 GASQSVSSSYLA 17 BADD197-1181 CDRL2 GASSRAT 18 BADD456-2928 CDRL2 AASTLQS 19 BADD466-3165 CDRL2 GASTRAT 20 BADD466-3166 DASSRAT Petition 870260065483, dated 03 / 07 / 2026, page 61 / 214 51 / 195 SEQ ID NO: Descrição* Sequência de aminoácidos CDRL2 21 BADD466-3167 CDRL2 DASNRAT 22 BADD 197-1181 CDRL3 QQYGSSPLT 23 BADD392-2234 CDRL3 QQYGSSPIT 24 BADD456-2919 VH EVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAM SWVRQAPGKGLEWVSAISGSGGSTYYADS VKGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 25 BADD466-3162 VH EVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAM SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 26 BADD466-3163 VH EVQLVESGGGLVQPRGSLRLSCAASGFTFS SYAM SWVRQAPGKGLEWVSAISGSGGSTYYADS VKGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 27 AM-1-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFK AGQS SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 28 AM-2-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFR QNAW SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 29 AM-3-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFM SGQT SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR Petition 870260065483, dated 03 / 07 / 2026, p. 62 / 214 52 / 195 SEQ ID NO: Descrição* Sequência de aminoácidos FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 30 AM-4-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFG AGQS SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 31 AM-5-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFS AQQA SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 32 AM-6-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFA RNAW SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 33 AM-7-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFR SQQA SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 34 AM-8-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFT TQQA SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 35 AM-9-VH EVQLVESGGGLVQPGGSLRLSCAASGFTFG GQQA SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD YGGNYFDYWGQGTLVTVSS 36 BADD197-1181 VL EIVLTQSPGTLSLSPGERATLSCRASQSVSS SYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSG SGSGT Petition 870260065483, dated 03 / 07 / 2026, p. 63 / 214 53 / 195 SEQ ID NO: Description* Amino acid sequence DFTLTISRLEPEDFAVYYCQQYGSSPLTFGG GTKV EIK 37 BADD412-2513 VL EIVLTQSPGTLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYGASSRATGIPDRFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 38 BADD456-2928 VL DIQMTQSPSSLSASVGDRVTITCRASQGISN YLAW YQQKPGKVPKLLIYAASTLQSGVPSRFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 39 BADD466-3164 VL EIVLTQSPATLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYGASSRATGIPDRFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVK IK 40 BADD466-3165 VL EIVLTQSPATLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYGASTRATGIPARFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 41 BADD466-3166 VL EIVLTQSPGTLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYDASSRATGIPDRFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 42 BADD466-3167 YLAW YQQKPGQAPRLLIYDASNRATGIPARFSGSG SGTD Petition 870260065483, dated 03 / 07 / 2026, p. 64 / 214 54 / 195 SEQ ID NO: Description* Amino acid sequence FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 44 BADD466-3169 VL EIVLTQSPATLSLSPGERATLSCGASQSVSS SYLA WYQQKPGLAPRLLIYDASSRATGIPDRFSGS GSGT DFTLTISRLEPEDFAVYYCQQYGSSPLTFGG GTKV EIK 45 BADD466-3170 VL EIVLTQSPATLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYDASNRATGIPARFSGSG SGTD FTLTISSLEPEDFAVYYCQQYGSSPLTFGGG TKVEI K 46 BADD466-3171 VL EIVLTQSPATLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYDASNRATGIPASFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPLTFGGG TKVEIK 47 BADD466-3172 VL EIVLTQSPGTLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYGASSRATGIPARFSGSG SGTD FTLTISRLEPEDFAVYYCQQYGSSPITFGGG TKVEI K 48 sequência consenso de CDRH1 1 X1X2X3X4X5S, em que: Xi é R, S, A, G, K, M ou T; X2éQ, S, A, G, Rou T; X3é N, Y, G ou Q; X4é A ou Q; e e X2 to A or S;and X3 is S or T 51 consensus sequence of CDRH1 4 X1X2QQAS, where: X is S, R, T or G; and X2 is A, S, T or G; Petition 870260065483, dated 03 / 07 / 2026, p. 65 / 214 55 / 195 SEQ ID NO: Description* Amino acid sequence 52 consensus sequence of CDRL1 X1ASQSVX2SSYLA, where Xi is R or G; and X2 is absent or is S 53 consensus sequence of CDRL2 XiASX2RAT, where: Xi is D or G; and X2 is N, S or T 54 consensus sequence of CDRL3 QQYGSSPX1T, where Xi is L or I 55 consensus sequence of VH EVQLVESGGGLVQPX1GSLRLSCAASGFTF X2X3 X4X5X6 S WVRX7APGKGLE WV S AIS GS GGS TY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAV YYCAKGGDYGGNYFDYWGQGTLVTVSS, where: Xi is G or R; X2 is R, S, A, G, K, M or T; X3 is Q, S, A, G, R or T; X4 is N, Y, G, or Q; X5 is A or Q; Xe is W, M, A, S, or T; and X7 is R or Q. 56 consensus sequence of VL EIVLTQSPX1TLSLSPGERATLSCX2ASQSVX3 SSYLAWYQQKPGX4APRLLIYX5ASX6RATGI PX7X8FSGSGSGTDFTLTISX9LEPEDFAVYYC QQYGSSPX10TFG GGTKVX11IK, where: Xi is A or G; X2 is R or G; Xs is absent or is S; X4 is Q or L; Xs is D or G; X6 is N, S, or T; X7 is A or D; X8 is S or R; X9 is R or S; X10 is L or I;and X11 is E or K 57 full-length IgG1 heavy chain pab2188 EVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAM SWVRRAPGKGLEWVSAISGSGGSTYYADSV KGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GGD; Petition 870260065483, dated 03 / 07 / 2026, p. 66 / 214 56 / 195 SEQ ID NO: Description* Amino acid sequence YG GN YF D YWGQ GTLVTVS S ASTKG PS VF PL APSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSG VHTFPAVLQSSGLYSLSSWTVPSSSLGTQT YICN VNHKPSNTKVDKRVEPKSCDKTHTCPPPCPA PELL GGPSVFLFPPKPKDTLMISRTPEVTCVWDV SHED P EVKFN WYVDGVEVH N AKTKP RE EQYN STY RW SVLTVLHQDWLNGKEYKCKVSNKALPAPI TIS KAKGQPREPQVYTLPPSREEMTKNQVSLTC LVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQK SLSLSPG 58 N297A heavy chain of full-length pab2188 IgG1 EVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTGGKCLVKGKGSTY DYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGP S VF LFPP KP KDTLM1S RTP E VTC VWD VS Η E DPEVKFNWYVNAKEQDVSTKWSTY VLTVL HQD WLN G KE YKC KVS N CAL P APIECTICAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YCTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSL 59SLdeSPGiaFull-length IgG4 S228P heavyweight of pab2188 EVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPCSRSTSESTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLS SWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGP PCPPCPAPEFLGGPSVFLFPPKPKDTLMISR Petition 870260065483, dated 03 / 07 / 2026, page 67 / 214 57 / 195 SEQ ID NO: Description* Amino acid sequence TPEVTCWVDVSQEDPEVQFNWYVDGVEV HNACTKPREEQFNSTYRWSVLTVLHQDWL NGKEYKCKVSNKGLPSSIEKTISKKGQPRE PQVYTLPPSQEMTKVKVKVKVKVKVL DIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSRLTVDKSRWQEGNVFSCSVMHEALHNH YTQKSLSLSLG 60 N297A heavy chain of full-length IgG1 of AM-1 YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKK TKVDKRVEPKSCDKTHTCPPCPAPELLGGP S VF LFPP KP KDTLM1S RTP E VTC VWD VS Η E DPEVKFNWYVDGVEVHNAKTKPREEQYAST YR WS VLTVL HQD WLN G KE YKC KVS N KAL PPEPQPPREKQYM NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 61 N297A heavy chain of full-length AM-2 IgG1 QNAWSWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTCGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGP S VF LFPP KP KDTLM 1S RTP E VTC VTCWD Η E DPEVKFNWYVDGVEVHNACTKPREEQYAST YR WS VLTVL HQD WLN G KE YKC KVS N CAL P APIEKTISCAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFYKFLYSKQLQQLTGW NVFSCSVMHEALHNHYTQKSLSLSPG 62 N297A heavy chain of full-length AM-3 IgG1 EVQLVESGGGLVQPGGSLRLSCAASGFTFM SGQTSWVRRAPGKGLEWVSAISGSGGSTY TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTCGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN Petition 870260065483, of 03 / 07 / 2026, p. 68 / 214 58 / 195 SEQ ID NO: Description* Amino Acid Sequence TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYAST YR WS VLTVL HQD WLN GKALK KVSKE N KVS APIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 63 heavy chain of N297A of full length IgG1 AM-4 EVQLVESGGGLVQPGGSLRLSCAASGFTFG AGQSSWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTGCLVKGKGKGSTY DYFPEPPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVKVWDVSHE DPEVKFNWYVDGVVHNAKT YPREQL WWLEQL GWLG KE YKC KVS N KAL P APIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 64 heavy chain NHYTQKSLSLSPG 64 heavy chain NHYTQKSLSLSPG complete with Ig297 AM-5 EVQLVESGGGLVQPGGSLRLSCAASGTFFS AQQASWVRRAPGKGLEWVSAISGSGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKK TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYAST YR WS VLTVL HQD WLN G KE YKC KVS N KAL P APIKTISKKGQPPEPCPAPELLGGP NQVSLTCLVKGFYPSDIAVEWESNGQPENN YCTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 65 N297A heavy chain of full-length AM-6 IgG1 RNAWSWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTCGVHTFPLQSS Petition 870260065483, of 03 / 07 / 2026, p. 69 / 214 59 / 195 SEQ ID NO: Description* Amino acid sequence GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSHE DPEVKFNWYVDGVVHNAKTQLWSTQLY WLN G KE YKC KVS N CAL P APIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSPG 66 cadeia of N2917 pesadari components of AM-7 EVQLVESGGGLVQPGGSLRLSCAASGFTFR SQQASWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVLAPSKSTGCTALGKCLVSGGSTY DYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVWDV SHEDPEVKFNWYVDGVEVHNAKTKPREEVLQH VLVLVVH KE YKC KVS N KALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPG 67 heavy chain of N297A comprimento AM-8 EVQLVESGGLVQPGGSLRLSCAASGFTFT TQQASWVRRAPGKGLEWVSAISGSGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK DYFPEPPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKK TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYAST YR WS VLTVL HQD WLN G KE YKC KVS N KAL P APIKTISKKGQPPEPCPAPELLGGP NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 68 N297A heavy chain of full-length AM-9 IgG1 GQQASWVRRAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKGGDYGGNYFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVK Petition 870260065483, of 03 / 07 / 2026, p. 70 / 214 60 / 195 SEQ ID NO: Descrição* Sequência de aminoácidos DYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSWTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVWDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYAST YR WS VLTVL HQD WLN G KE YKC KVS N KAL P APIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPG 69 sequences of video games comprimento completo BADD466-3171 HKPSNT KVDKRVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCWVDVSSHED P EVKFN WYVDGVEVH N AKTKP RE EQYN STY RWSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPG 71 Glm3 allotype of human IgG1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPPVTVSWNSGALTSGVHTFPAVLQSSG LYSL KVDKRVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCWVDVSHED P EVKFN WYVDGVEVH N AKTKP RE EQYN STY RWSVLTVLHQDWLNGKEYKCKVSNKALPA PRACTICE QVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK 72 N297A of IgG1 (with C-terminal lysine) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDKGN YFPEPTVSWNSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTYICNVNHKPSNT KVDKRVEPKSCDKTHTCPPCPPAPELLGGPS Petition 870260065483, of 03 / 07 / 2026, p. 71 / 214 61 / 195 SEQ ID NO: Descrição* Sequência de aminoácidos VFLFPPKPKDTLMISRTPEVTCVWDVSHED P EVKFN WYVDGVEVH N AKTKP RE EQYASTY RWSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPG 73 N297A of IgG1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTYICNVNHKPSNT KVDKRVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVWDVSHED P EVKFN WYVDGVEVH N AKTKP RE EQYASTY RWSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSC SVMHEALHNHYTQKSLSLSPGK 74 S228P of IgG4 (sem lisina C-terminal) ASTKGPSVFPLAPCSRSTSESTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPPCPAPEFLGGPSVFL FPP KP KDTLM IS RTP EVTCVWDVS QE DP EV QFNWYVDGVEVHNAKTKPREEQFNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE KTISKAKGQPREPQVYTLLPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 75 S228P IgG4 ASTKGPSVFPLAPCSRSTSESTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPPCPAPEFLGGPSVFL FPP KP KDTLM IS RTP EVTCVWDVS QE DP EV QFNWYVDGVEVHNAKTKPREEQFNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE KTISKAKGQPREPQVYTLPPSQEEMTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVMHEALHNHYTQKSLSLSLGK 76 IGKC*01 allotype Km3 of constant region RTVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQD Petition 870260065483, dated 03 / 07 / 2026, page 72 / 214 62 / 195 SEQ ID NO: Descrição* Sequence of amino acids from the human life group SKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 77 IGKC*01 km3 of constant região of the human life group (commuted to T109S) RSVAAPSVFIFPPSDEQLKSGTASWCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 84 IGHV3-23*04 QVQLVESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVSAISGSGGSTY 86 IGKV3-11*01 EIVLTQSPATLSLSPGERATLSCRASQSVSS YLAW YQQKPGQAPRLLIYDASNRATGIPARFSGSG SGTD FTLTIS SLEPEDFAVYYCQQRSNWP 87 IGKV3-20*01 EIVLTQSPGTLSLSPGERATLSCRASQSVSS SYLAWYQQKPGQAPRLLIYGASSRATGIPDR SSP *The CDRs for heavy chains are defined according to the MacCallum numbering system, and the CDRs for light chains are defined according to the Rabat numbering system. TABLE 2. CHAIN-BOUND AMINO ACID SEQUENCES EXAMPLE CAPTIVE ANTI-TIM-3 ANTIBODIES WEIGHT. VH CDRH1* SEQ ID NO: CDRH2* SEQ ID NO: CDRH3* SEQ ID NO: BADD456- 2919 SSYAMS 1 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 BADD466- SSYAMS 1 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 Petition 870260065483, dated 03 / 07 / 2026, page 73 / 214 63 / 195 3162 BADD466- 3163 SSYAMS 1 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-1 KAGQSS 4 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-2 RQNAWS 5 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-3 MSGQTS 6 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-4 GAGQSS 7 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-5 SAQQAS 8 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-6 ARNAWS 9 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-7 RSQQAS 10 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-8 TTQQAS 11 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 AM-9 GGQQAS 12 WVSAISGSGGSTY 2 AKGGDYGGNYFD 3 *Defined according to the MacCallum numbering system. TABLE 3. LIGHT CHAIN ​​CDR AMINO ACID SEQUENCES EXEMPLARY TYPES OF ANTI-TIM-3 ANTIBODIES. VL CDRL1* SEQ ID NO: CDRL2* SEQ ID NO: CDRL3* SEQ ID NO: BADD197- 1181 RASQSVSSSYLA 13 GASSRAT 17 QQYGSSPLT 22 BADD412- 2513 RASQSVSSYLA 14 GASSRAT 17 QQYGSSPLT 22 BADD456- 2928 RASQGISNYLA 15 AASTLQS 18 QQYGSSPLT 22 BADD466- 3164 RASQSVSSYLA 14 GASSRAT 17 QQYGSSPLT 22 BADD466- 3165 RASQSVSSYLA 14 GASTRAT 19 QQYGSSPLT 22 BADD466- 3166 RASQSVSSYLA 14 DASSRAT 20 QQYGSSPLT 22 BADD466- 3167 RASQSVSSYLA 14 DASNRAT 21 QQYGSSPLT 22 BADD466- 3168 RASQSVSSYLA 14 DASNRAT 21 QQYGSSPLT 22 Petition: 870260065483, on 03 / 07 / 2026, page. 74 / 214 64 / 195 BADD466- 3169 GASQSVSSSYLA 16 DASSRAT 20 QQYGSSPLT 22 BADD466- 3170 RASQSVSSYLA 14 DASNRAT 21 QQYGSSPLT 22 BADD466- 3171 RASQSVSSYLA 14 DASNRAT 21 QQYGSSPLT 22 BADD466- 3172 RASQSVSSYLA 14 GASSRAT 17 QQYGSSPIT 23 *Definition of the record from the Kabat number system. TABELA 4. ANTI-TIM-3 EXEMPLIFICATIVOS. Antibody Heavy chain variable region SEQ ID NO: Light chain variable region SEQ ID NO: pab2085 BADD456-2919 24 BADD197-1181 36 pab2088 BADD456-2919 24 BADD456-2928 38 pab2173 BADD466-3163 26 BADD466-3167 42 pab2174 BADD456-2919 24 BADD466-3167 42 pab2175 BADD456-2919 24 BADD466-3171 46 pab2176 BADD456-2919 24 BADD466-3168 43 pab2177 BADD466-3163 26 BADD466-3168 43 pab2178 BADD466-3163 26 BADD466-3171 46 pab2179 BADD466-3163 26 BADD466-3166 41 pab2180 BADD456-2919 24 BADD466-3166 41 pab2181 BADD466-3162 25 BADD466-3164 39 pab2182 BADD456-2919 24 BADD466-3172 47 pab2183 BADD466-3162 25 BADD466-3165 40 pab2184 BADD466-3163 26 BADD466-3172 47 pab2185 BADD466-3162 25 BADD412-2513 37 pab2186 BADD466-3162 25 BADD466-3170 45 pab2187 BADD466-3162 25 BADD466-3169 44 pab2188 BADD466-3162 25 BADD466-3171 46 pab2189 BADD466-3162 25 BADD466-3167 42 pab2190 BADD466-3162 25 BADD466-3166 41 Petition 870260065483, dated 03 / 07 / 2026, page 75 / 214 65 / 195 pab2191 BADD466-3162 25 BADD466-3168 43 pab2192 BADD466-3162 25 BADD466-3172 47 AM-1 AM-1-VH 27 BADD466-3171 46 AM-2 AM-2-VH 28 BADD466-3171 46 AM-3 AM-3-VH29 BADD466-3171 46 AM-4 AM-4-VH 30 BADD466-3171 46 AM-5 AM-5-VH 31 BADD466-3171 46 AM-6 AM-6-VH 32 BADD466-3171 46 AM-7 AM-7-VH 33 BADD466-3171 46AM-8 AM-8-VH 34 BADD466-3171 46 AM-9 AM-9-VH 35 BADD466-3171 46 TABLE 5. GENES OF CLOSEST GERMLINE RELATED. Heavy chain or light chain variable region. Closest germline gene. SEQ ID NO for germline gene: BADD456-2919 VH IGHV3-23*04 84 BADD466-3162 VH IGHV3-23*04 84 BADD466-3163 VH IGHV3-23*04 84 AM-1-VH IGHV3-23*04 84 AM-2-VH IGHV3-23*04 84 AM-3-VH IGHV3-23*04 84 AM-4-VH IGHV3-23*04 84 AM-5-VH IGHV3-23*04 84 AM-6-VH IGHV3-23*04 84 AM-7-VH IGHV3-23*04 84 AM-8-VH IGHV3-23*04 84 AM-9-VH IGHV3-23*04 84 BADD197-1181 VL IGKV3-20*01 87 BADD412-2513 VL IGKV3-20*01 87 BADD456-2928 VL IGKV1-27*01 85 BADD466-3164 VL IGKV3-20*01 87 Petition 870260065483, dated 03 / 07 / 2026, page 76 / 214 66 / 195 BADD466-3165 VL IGKV3-20*01 87 BADD466-3166 VL IGKV3-20*01 87 BADD466-3167 VL IGKV3-11*01 86 BADD466-3168 VL IGKV3-20*01 87 BADD466-3169 VL IGKV3D-20*01 88 BADD466-3170 VL IGKV3-11*01 86 BADD466-3171 VL IGKV3-11*01 86 BADD466-3172 VL IGKV3-20*01 87 TABLE 6. EXEMPLARY SEQUENCES OF TIM-3. SEQ ID NO: Description* Amino acid sequence 78 Immature human TIM-3 protein (Q8TDQ0-1) MF SHLPFDC VLLLLLLLLTRS SEVE YRAEVGQNAYLPCFYTPAAPGNLVPVCWGK GACPVFECGNWLRTDERDVNYWTSRYWL NGDFRKGDVSLTIENVTLADSGIYCCRIQIPGI MNDEKFNLKLVIKPAKVTPAPTRQRDFTAAF PRMLTTRGHGPAETQTLGSLPDINLTQISTLA NELRDSRLANDLRDSGATIRIGIYIGAGICAGL ALALIFGALIFKWYSHSKEKIQNLSLISLANLP PSGLANAVAEGIRSEENIYTIEENVYEVEEPN EYYCYVSSRQQPSQPLGCRFAMP 79 Mature human TIM-3 protein SEVEYRAEVGQNAYLPCFYTPAAPGNLVPV CWGKGACPVFECGNWLRTDERDVNYWTS RYWLNGDFRKGDVSLTIENVTLADSGIYCCR IQIPGIMNDEKFNLKLVIKPAKVTPAPTRQRD FTAAFPRMLTTRGHGPAETQTLGSLPDINLT QISTLANELRDSRLANDLRDSGATIRIGIYIGA GICAGLALALIFGALIFKWYSHSKEKIQNLSLI SLANLPPSGLANAVAEGIRSEENIYTIEENVY EVEEPNEYYCYVSSRQQPSQPLGCRFAMP 101 F40A of human TIM-3 SEVEYRAEVGQNAYLPCFYTPAAPGNLVPV CWGKGACPVAECGNWLRTDERDVNYWTS RYWLNGDFRKGDVSLTIENVTLADSGIYCCR IQIPGIMNDEKFNLKLVIKPAKVTPAPTRQRD FTAAFPRMLTTRGHGPAETQTLGSLPDINLT QISTLANELRDSRLANDLRDSGATIRIGIYIGA GICAGLALALIFGALIFKWYSHSKEKIQNLSLISLANLPPSGLANAVAEGIRSEENIYTIEENVY EVEEPNEYYCYVSSRQQPSQPLGCRFAMP 102 Fragment of human TIM-3 SEVEYRAEVGQNAYLPCFYTPAAPGNLVPV CWGKGACPVFECGNWLRTDERDVNYWTS RYWLNGDFRKGDVSLTIENVTLADSGIYCCR Petition 870260065483, dated 03 / 07 / 2026, page 77 / 214 67 / 195 IQIPGIMNDEKFNLKLVIKPAKVTPAPTRQRD FTAAFPRMLTTRGHGPAETQTLGSLPDINLT QISTLANELRDSRLANDLRDSGATIR ​​93 TIM-3 epitope PVFECGN 94 TIM-3 epitope VCWGKGACPVFECGNWL 95 TIM-3 epitope RIQIPGIMND 96 TIM-3 epitope RIQIPGIMNDEKFNLKL 97 TIM-3 epitope EKFNLKL 98 TIM-3 epitope PAAPGNLVP 99 TIM-3 epitope GKGACPVFE 100 TIM-3 epitope DFTAAFPR

[0140] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VH domain comprising one, two, or all three CDRs of a VH domain presented in Table 1 thereto. In certain embodiments, the antibody comprises CDRH1 of one of the VH domains presented in Table 1. In certain embodiments, the antibody comprises CDRH2 of one of the VH domains presented in Table 1. In certain embodiments, the antibody comprises CDRH3 of one of the VH domains presented in Table 1.

[0141] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VL domain comprising one, two, or all three CDRs of a VL domain disclosed in Table 1 therein. In certain embodiments, the antibody comprises CDRL1 of one of the VL domains presented in Table 1. In certain embodiments, the antibody comprises CDRL2 of one of the VL domains presented in Table 1. In certain embodiments, the antibody comprises CDRL3 of one of the VL domains presented in Table 1.

[0142] In certain modalities, the CDRs of an antibody can be determined Petition 870260065483, dated 03 / 07 / 2026, page 78 / 214 68 / 195 according to MacCallum RM et al., (1996) J Mol Biol 262: 732-745, incorporated herein by reference in its entirety). See also, for example, Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Dtibel, eds., Chapter 31, pages 422-439, Springer-Verlag, Berlin (2001), incorporated herein by reference in its entirety. In certain embodiments, the heavy chain CDRs of an antibody are determined according to MacCallum and the light chain CDRs of an antibody are determined according to a different method.

[0143] In certain embodiments, the CDRs of an antibody may be determined according to Rabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Rabat et al., Sequences of protein of immunological interest (1991), each of which is incorporated herein by reference in its entirety. In certain embodiments, the light chain CDRs of an antibody are determined according to Rabat and the heavy chain CDRs of an antibody are determined according to MacCallum (above).

[0144] In certain embodiments, the CDRs of an antibody can be determined according to the Chothia numbering scheme, which refers to the location of immunoglobulin structural loops (see, for example, Chothia C & Lesk AM, (1987), J Mol Biol 196: 901-917; Al-Lazikani B et al., (1997) J Mol Biol 273: 927-948; Chothia C et al., (1992) J Mol Biol 227: 799-817; Tramontano A et al., (1990) J Mol Biol 215(1): 175-182; and US Patent No. 7,709,226, all of which are incorporated herein by reference in their entirety). Typically, when using the Kabat numbering convention, the CDRH1 loop of Chothia is present in heavy chain amino acids 26 to 32, 33, or 34; the CDRH2 loop of Chothia is present in heavy chain amino acids 52 to 56; and the CDRH3 loop of Chothia is present in heavy chain amino acids 95 to 102, while the CDRL1 loop of Chothia is present in light chain amino acids 24 to 34. Petition 870260065483, dated 03 / 07 / 2026, page 79 / 214 The 69 / 195 Chothia CDRL2 loop is present in light chain amino acids 50 to 56, and the Chothia CDRL3 loop is present in light chain amino acids 89 to 97. The end of the Chothia CDRH1 loop, when numbered using the Kabat numbering convention, varies between H32 and H34 depending on the loop length (this is because the Kabat numbering scheme positions the insertions at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34).

[0145] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises the Chothia VH CDRs of an HV disclosed in Table 1 therein. In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises the Chothia VL CDRs of an VL disclosed in Table 1 therein. In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises the Chothia VH CDRs and the Chothia VL CDRs of an antibody disclosed in Table 1 therein. In certain embodiments, antibodies that bind specifically to TIM-3 (e.g., human TIM-3) comprise one or more CDRs, where the Chothia and Kabat CDRs have the same amino acid sequence.In certain embodiments, the present disclosure provides a single antibody that binds specifically to TIM-3 (e.g., human TIM-3) and comprises combinations of Kabat CDRs and Chothia CDRs.

[0146] In certain embodiments, the CDRs of an antibody can be determined according to the IMGT numbering system as described in Lefranc MP, (1999) The Immunologist 7: 132-136 and Lefranc MP et al., (1999) Nucleic Acids Res 27: 209-212, each of which is incorporated herein by reference. Petition 870260065483, dated 03 / 07 / 2026, p. 80 / 214 70 / 195 reference in its entirety). According to the IMGT numbering scheme, CDRH1 is in positions 26 to 35, CDRH2 is in positions 51 to 57, CDRH3 is in positions 93 to 102, CDRL1 is in positions 27 to 32, CDRL2 is in positions 50 to 52, and CDRL3 is in positions 89 to 97.

[0147] In certain embodiments, the present disclosure provides antibodies that bind specifically to TIM-3 (e.g., human TIM-3) and comprise CDRs of an antibody disclosed in Table 1 herein, as determined by the IMGT numbering system, for example, as described in Lefranc MP (1999) supra and Lefranc MP et al., (1999) supra.

[0148] In certain embodiments, the CDRs of an antibody can be determined according to the AbM numbering scheme, which refers to AbM hypervariable regions, representing a compromise between Kabat CDRs and Chothia structural loops and used by the Oxford Molecular (Oxford Molecular Group, Inc.) antibody modeling software AbM, incorporated herein by reference in its entirety. In a particular embodiment, the present disclosure provides antibodies that specifically bind to TIM-3 (e.g., human TIM-3) and comprise CDRs of an antibody disclosed in Table 1 herein as determined by the AbM numbering scheme.

[0149] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a variable heavy chain region comprising the amino acid sequences CDRH1, CDRH2, and CDRH3 of a VH domain presented in SEQ ID NO: 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35, and a variable light chain region comprising the amino acid sequences CDRL1, CDRL2, and CDRL3 of a VL domain presented in SEQ ID NO: 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, or 47, wherein each CDR is defined in accordance with the MacCallum definition, the Kabat definition, the definition of Petition 870260065483, dated 03 / 07 / 2026, page 81 / 214 71 / 195 Chothia, the combination of Kabat's definition and Chothia's definition, the IMGT numbering system, or CDR's AbM definition.

[0150] In certain embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises: (a) a CDRH1 comprises the amino acid sequence X1X2X3X4X5S (SEQ ID NO: 48), where X1 is R, S, A, G, K, M, or T. X2 is Q, S, A, G, R, or T. X3 is N, Y, G, or Q. X4 is A or Q, and X5 is W, M, A, S, or T; and / or (b) a CDRH2 comprises the amino acid sequence of WVSAISGSGGSTY (SEQ ID NO: 2); and / or (c) a CDRH3 comprises AKGGDYGGNYFD (SEQ ID NO: 3); and / or (d) a CDRL1 comprises the amino acid sequence of the amino acid sequence of X1ASQSVX2SSYLA (SEQ ID NO: 52), where X1 is R or G; and X2 is absent or is S; and / or (d) a CDRL2 comprises the amino acid sequence of X1ASX2RAT ​​(SEQ ID NO: 53), where X1 is D or G; and X2 is N, S, or T; and / or (and) a CDRL3 comprises the amino acid sequence of QQYGSSPX1T (SEQ ID NO: 54), where X1 is L or I. Petition 870260065483, dated 03 / 07 / 2026, p. 82 / 214 72 / 195

[0151] In certain embodiments, CDRH1 comprises the amino acid sequence X1X2NAWS (SEQ ID NO: 49), where: X1 is R or A; and X2 is Q or R. In certain embodiments, CDRH1 comprises the amino acid sequence X1X2GQX3S (SEQ ID NO: 50), where: X1 is K, M, or G; X2 is A or S; and X3 is S or T. In certain embodiments, CDRH1 comprises the amino acid sequence X1X2QQAS (SEQ ID NO: 51), where: X1 is S, R, T, or G; X2 is A, S, T, or G. In certain embodiments, CDRH1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 and 4 to 12. In certain embodiments, CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 13 to 16. In certain embodiments, CDRL2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 21. In certain embodiments, CDRL3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 and 23.

[0152] In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VH domain comprising the amino acid sequences CDRH1, CDRH2 and CDRH3 presented in the SEQ ID NOs: 1, 2 and 3; 4, 2 and 3; 5, 2 and 3; 6, 2 and 3; 7, 2 and 3; 8, 2 and 3; 9, 2 and 3; 10, 2 and 3; 11, 2 and 3; or 12, 2 and 3 respectively. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VH domain comprising the amino acid sequences CDRH1, CDRH2, and CDRH3 presented in SEQ ID NOs: 1, 2, and 3, respectively.In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VH domain comprising the amino acid sequences CDRH1, CDRH2, and CDRH3 presented in SEQ ID NOs: 5, 2, and 3, respectively. In certain embodiments, the present invention provides an isolated antibody that binds. Petition 870260065483, dated 03 / 07 / 2026, p. 83 / 214 73 / 195 specifically TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VH domain comprising the amino acid sequences CDRH1, CDRH2, and CDRH3 presented in SEQ ID NOs: 9, 2, and 3, respectively.

[0153] In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VL domain comprising the amino acid sequences CDRL1, CDRL2 and CDRL3 presented in SEQ ID NOs: 13, 17 and 22; 14, 17 and 22; 15, 18 and 22; 14, 19 and 22; 14, 20 and 22; 14, 21 and 22; 16, 20 and 22; or 14, 17 and 23, respectively. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a VL domain comprising the amino acid sequences CDRL1, CDRL2, and CDRL3 presented in SEQ ID NOs: 14, 21, and 22, respectively.

[0154] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a variable heavy chain region comprising the CDRH1, CDRH2 and CDRH3 regions and a variable light chain region comprising the CDRL1, CDRL2 and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 regions comprise the amino acid sequences presented in the SEQ ID NOs: 1, 2, 3, 14, 21 and 22; 4, 2, 3, 14, 21 and 22; 5, 2, 3, 14, 21 and 22; 6, 2, 3, 14, 21 and 22; 7, 2, 3, 14, 21 and 22; 8, 2, 3, 14, 21 and 22; 9, 2, 3, 14, 21 and 22; 10, 2, 3, 14, 21 and 22; 11, 2, 3, 14, 21 and 22; or 12, 2, 3, 14, 21 and 22, respectively.In certain embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a variable heavy chain region comprising the CDRH1, CDRH2, and CDRH3 regions and a variable light chain region comprising the CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, regions... Petition 870260065483, dated 03 / 07 / 2026, page 84 / 214 74 / 195 CDRL2 and CDRL3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, 14, 21, and 22, respectively. In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a variable heavy chain region comprising the CDRH1, CDRH2, and CDRH3 regions and a variable light chain region comprising the CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences presented in SEQ ID NOs: 5, 2, 3, 14, 21, and 22, respectively.In certain embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a variable heavy chain region comprising the CDRH1, CDRH2, and CDRH3 regions and a variable light chain region comprising the CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences presented in the SEQ ID NOs: 9, 2, 3, 14, 21, and 22.

[0155] In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region comprising an amino acid sequence of SEQ ID NO: 55. In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence shown in SEQ ID NO: 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35. In certain embodiments, Petition 870260065483, dated 03 / 07 / 2026, p. 85 / 214 75 / 195 The antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 24. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 25. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 26. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 27. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 28. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 29.In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 30. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 31. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 32. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 33. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 34. In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence presented in SEQ ID NO: 35.In certain embodiments, the N-terminal glutamate (E) residue of a variable heavy chain region of an antibody as described in this document is replaced by a pyroglutamate (pE) residue.

[0156] In certain embodiments, the present invention provides an iso antibody Petition 870260065483, dated 03 / 07 / 2026, p. 86 / 214 76 / 195 side that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable light chain region comprising an amino acid sequence of SEQ ID NO: 56. In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable light chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence shown in SEQ ID NO: 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47.In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 36. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 37. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 38. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 39. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 40. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence presented in SEQ ID NO: 41.In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 42. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 43. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown. Petition 870260065483, dated 03 / 07 / 2026, page 87 / 214 77 / 195 in SEQ ID NO: 44. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 45. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 46. In certain embodiments, the antibody comprises a variable light chain region having the amino acid sequence shown in SEQ ID NO: 47. In certain embodiments, the N-terminal glutamate (E) residue of a variable light chain region of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[0157] In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region comprising an amino acid sequence of SEQ ID NO: 55 and a variable light chain region comprising an amino acid sequence of SEQ ID NO: 56.In certain embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to the amino acid sequence shown in SEQ ID NO: 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 and a variable light chain region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (for example, at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%) identical to the amino acid sequence shown in SEQ ID NO: 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47.In certain embodiments, the antibody comprises a variable heavy chain region having the amino acid sequence shown in SEQ ID NO: 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 and a variable light chain region having the amino acid sequence shown. Petition 870260065483, dated 03 / 07 / 2026, p. 88 / 214 78 / 195 in SEQ ID NO: 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region that have the amino acid sequences presented in SEQ ID NO: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 24 and 36, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 24 and 38, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 26 and 42, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 24 and 42, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 24 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 24 and 43, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 26 and 43, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in... Petition 870260065483, dated 03 / 07 / 2026, page 89 / 214 79 / 195 SEQ ID NO: 26 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region that have the amino acid sequences presented in SEQ ID NO: 26 and 41, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region that have the amino acid sequences presented in SEQ ID NO: 24 and 41, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region that have the amino acid sequences presented in SEQ ID NO: 25 and 39, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region that have the amino acid sequences presented in SEQ ID NO: 24 and 47, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 40, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 26 and 47, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 37, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 45, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 44, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 46, respectively. In certain embodiments... Petition 870260065483, dated 03 / 07 / 2026, pp. 90 / 214 80 / 195 The antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 42, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 41, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 43, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 25 and 47, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 27 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 28 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 29 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 30 and 46, respectively.In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 31 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences presented in SEQ ID NO: 32 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and... Petition 870260065483, dated 03 / 07 / 2026, page 91 / 214 81 / 195 a variable light chain region having the amino acid sequences shown in SEQ ID NO: 33 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences shown in SEQ ID NO: 34 and 46, respectively. In certain embodiments, the antibody comprises a variable heavy chain region and a variable light chain region having the amino acid sequences shown in SEQ ID NO: 35 and 46, respectively. In certain embodiments, the N-terminal glutamate (E) residue of a variable heavy chain region of an antibody as described herein is replaced by a pyroglutamate (pE) residue and / or the N-terminal glutamate (E) residue of a variable light chain region of the antibody is replaced by a pyroglutamate (pE) residue.

[0158] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region having an amino acid sequence derived from a human IGHV3-23 germline sequence (e.g., IGHV3-23*04, for example, which has the amino acid sequence of SEQ ID NO: 84). One or more regions selected from framework 1, framework 2, framework 3, CDRH1 and CDRH2 (e.g., two, three, four or five of these regions) may be derived from a human IGHV3-23 germline sequence (e.g., IGHV3-23*04, for example, which has the amino acid sequence of SEQ ID NO: 84). In one embodiment, structure 1, structure 2, structure 3, CDRH1, and CDRH2 are all derived from a germline sequence of human IGHV3-23 (e.g., IGHV3-23*04, for example, which has the amino acid sequence SEQ ID NO: 84).

[0159] In certain embodiments, the present disclosure provides an antibody Petition 870260065483, dated 03 / 07 / 2026, p. 92 / 214 82 / 195 isolate that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable light chain region having an amino acid sequence derived from a human germline sequence selected from the group consisting of IGKV1-27 (e.g., IGKV1-27*01, for example, having the amino acid sequence of SEQ ID NO: 85), IGKV3-11 (e.g., IGKV311*01, for example, having the amino acid sequence of SEQ ID NO: 86), IGKV320 (e.g., IGKV3-20*01, for example, having the amino acid sequence of SEQ ID NO: 87) and IGKV3D-20 (e.g., IGKV3D-20*01, for example, having the amino acid sequence of SEQ ID NO: 88).One or more selected regions from framework 1, framework 2, framework 3, CDRL1, and CDRL2 (for example, two, three, four, or five of these regions) may be derived from a human germline sequence selected from the group consisting of IGKV1-27 (for example, IGKV1-27*01, for example, having the amino acid sequence of SEQ ID NO: 85), IGKV3-11 (for example, IGKV3-11*01, for example, having the amino acid sequence of SEQ ID NO: 86), IGKV3-20 (for example, IGKV3-20*01, for example, having the amino acid sequence of SEQ ID NO: 87), and IGKV3D-20 (for example, IGKV3D-20*01, for example, having the amino acid sequence of SEQ ID NO: 88).In one embodiment, structure 1, structure 2, structure 3, CDRL1, and CDRL2 are all derived from a selected human germline sequence from the group consisting of IGKV1-27 (e.g., IGKV1-27*01, for example, having the amino acid sequence SEQ ID NO: 85), IGKV3-11 (e.g., IGKV3-11*01, for example, having the amino acid sequence SEQ ID NO: 86), IGKV3-20 (e.g., IGKV3-20*01, for example, having the amino acid sequence SEQ ID NO: 87), and IGKV3D-20 (e.g., IGKV3D-20*01, for example, having the amino acid sequence SEQ ID NO: 88).

[0160] In certain embodiments, the present disclosure provides an antibody Petition 870260065483, dated 03 / 07 / 2026, p. 93 / 214 83 / 195 isolate that specifically binds to TIM-3 (e.g., human TIM-3), comprising a variable heavy chain region having an amino acid sequence derived from a germline sequence of human IGHV3-23 (e.g., IGHV3-23*04, for example, which has the amino acid sequence SEQ ID NO: 84), and a variable light chain region having an amino acid sequence derived from a human germline sequence selected from the group consisting of IGKV1-27 (e.g., IGKV1-27*01, for example, which has the amino acid sequence SEQ ID NO: 85), IGKV3-11 (e.g., IGKV3-11*01, for example, which has the amino acid sequence SEQ ID NO: 86), IGKV3-20 (e.g., IGKV3-20*01, for example, which has the amino acid sequence SEQ ID NO: 87) and IGKV3D-20 (for example, IGKV3D-20*01, for example, which has the amino acid sequence SEQ ID NO: 88).

[0161] In certain embodiments, the present disclosure provides an isolated antibody that cross-competes for binding to TIM-3 (e.g., human TIM-3) with an antibody comprising the variable region heavy and light chain amino acid sequences presented in the SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46, respectively.

[0162] In certain embodiments, the present disclosure provides an isolated antibody that binds to the same epitope or to an overlapping epitope of TIM-3 (for example, a human TIM-3 epitope) as an antibody described in this document, for example, an antibody comprising the heavy and light chain variable region amino acid sequences presented in the SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 Petition 870260065483, dated 03 / 07 / 2026, pp. 94 / 214 84 / 195 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46, respectively. In certain embodiments, the epitope of an antibody can be determined, for example, by NMR spectroscopy, surface plasmon resonance (BIAcore®), X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled to mass spectrometry (e.g., liquid chromatography-electron scatter mass spectrometry), array-based oligopeptide screening assays and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, crystallization can be performed using any of the methods known in the art (e.g., Giege R et al.)., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; Chayen NE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303, all of which are incorporated herein by reference in their entirety). Antibody:antigen crystals can be studied using well-known X-ray diffraction techniques and can be refined using computer software such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985) volumes 114 and 115, eds Wyckoff HW et al.,; US Patent Application No. 2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323, all of which are incorporated herein by reference in their entirety). Mutagenesis mapping studies can be performed using any method known to those skilled in the art. See, for example, Champe M et al., (1995) supra and Cunningham BC & Wells JA (1989) supra for a description of mutagenesis techniques, including alanine screening mutagenesis techniques. In one specific embodiment, the epitope of an antibody is determined with the. Petition 870260065483, dated 03 / 07 / 2026, pp. 95 / 214 85 / 195 use of alanine screening mutagenesis studies. Additionally, antibodies that recognize and bind to identical or overlapping epitopes of TIM-3 (e.g., human TIM-3) can be identified using routine techniques, such as an immunoassay, for example, by showing the ability of an antibody to block the binding of another antibody to a target antigen, i.e., a competitive binding assay. Competitive binding assays can also be used to determine whether two antibodies have similar binding specificity for an epitope. Competitive binding can be determined in an assay where the immunoglobulin under test inhibits the specific binding of a reference antibody to a common antigen, such as TIM-3 (e.g., human TIM-3).Numerous types of competitive binding assays are known, for example: direct or indirect solid-phase radioimmunoassay (RIA), direct or indirect solid-phase enzyme immunoassay (EIA), sandwich competition assay (see Stahli C et al., (1983) Methods Enzymol 9: 242-253); direct solid-phase biotin-avidin EIA (see Kirkland TN et al., (1986) J Immunol 137: 3614-3619); direct solid-phase identified assay, direct solid-phase identified sandwich assay (see Harlow E & Lane D, (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Press); direct solid-phase label RIA using I-125 label (see Morel GA et al., (1988) Mol Immunol 25(1): 7-15); EIA of biotin-avidin direct solid phase (see Cheung RC et al., (1990) Virology 176: 546-552); and RIA with direct marker (see Moldenhauer G et al.), (1990) Scand J Immunol 32: 77-82), each of which is incorporated herein by reference in its entirety. Typically, such an assay involves the use of purified antigen (e.g., TIM-3, such as human TIM-3) bound to a solid surface or cells bearing either, an unidentified test immunoglobulin, and an identified reference immunoglobulin. Competitive inhibition can be measured by determining the amount of marker bound to the solid surface or cells in the presence of the test immunoglobulin. Petition 870260065483, dated 03 / 07 / 2026, p. 96 / 214 86 / 195 Usually, the test immunoglobulin is present in excess. Usually, when a competing antibody is present in excess, it will inhibit the specific binding of a reference antibody to a common antigen by at least 50 to 55%, 55 to 60%, 60 to 65%, 65 to 70%, 70 to 75% or more. A competitive binding assay can be set up in a large number of formats using identified antigen or identified antibody. In a common version of this assay, the antigen is immobilized in a 96-well plate. The ability of unidentified antibodies to block the binding of identified antibodies to the antigen is then measured using radioactive or enzymatic markers. For more details see, for example, Wagener C et al., (1983) J Immunol 130: 2308-2315; Wagener C et al., (1984) J Immunol Methods 68: 269-274; Kuroki M et al., (1990) Cancer Res 50: 4872-4879; Kuroki M et al., (1992) Immunol Invest 21: 523-538; Kuroki M et al., (1992) Hybridoma 11: 391-407 and Antibodies: A Laboratory Manual, Ed Harlow E & Lane D editors supra, pages 386-389, all of which are incorporated into this document by reference in their entirety.

[0163] In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, or 68. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 57. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 58. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 59. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 60. Petition 870260065483, dated 03 / 07 / 2026, p. 97 / 214 87 / 195 In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 61. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 62. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 63. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 64. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 66. In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 67.In certain embodiments, the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 68. In certain embodiments, the N-terminal glutamate (E) residue of a heavy chain of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[0164] In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a light chain comprising the amino acid sequence shown in SEQ ID NO: 69. In certain embodiments, the N-terminal glutamate (E) residue of a light chain of an antibody as described herein is replaced by a pyroglutamate (pE) residue.

[0165] In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence Petition 870260065483, dated 03 / 07 / 2026, pp. 98 / 214 88 / 195 acids of SEQ ID NO: 57; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 58; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 59; and a light chain comprising the amino acid sequence of SEQ ID NO: 69.In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 60; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 61; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 62; and a light chain comprising the amino acid sequence of SEQ ID NO: 69.In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 63; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that binds... Petition 870260065483, dated 03 / 07 / 2026, p. 99 / 214 89 / 195 specifically TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 64; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 65; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 66; and a light chain comprising the amino acid sequence of SEQ ID NO: 69.In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 67; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 68; and a light chain comprising the amino acid sequence of SEQ ID NO: 69. In certain embodiments, the N-terminal glutamate (E) residue of a heavy chain of an antibody as described herein is replaced by a pyroglutamate (pE) residue and / or the N-terminal glutamate (E) residue of a light chain of the antibody is replaced by a pyroglutamate (pE) residue.

[0166] Any constant Ig region can be used in the antibodies disclosed in this document. In certain embodiments, the Ig region is a molecule Petition 870260065483, dated 03 / 07 / 2026, pp. 100 / 214 90 / 195 of human immunoglobulin IgG, IgE, IgM, IgD, IgA or IgY, any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or any subclass (e.g., IgG2a and IgG2b) of immunoglobulin molecule.

[0167] In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 70, 71, 72, 73, 74 or 75. In certain embodiments, the present invention provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 76 or 77.

[0168] In certain embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of an antibody described in this document (e.g., CH2 domain (residues 231 to 340 of human IgG1) and / or CH3 domain (residues 341 to 447 of human IgG1) and / or the hinge region, numbered according to the EU numbering system, to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding and / or antigen-dependent cellular cytotoxicity.

[0169] In certain embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain) so that the number of cysteine ​​residues in the hinge region is altered (e.g., increased or decreased) as described, for example, in U.S. Patent No. 5,677,425, which is incorporated herein by reference in its entirety. The number of cysteine ​​residues in the hinge region of the CH1 domain may be altered, for example, to facilitate assembly. Petition 870260065483, dated 03 / 07 / 2026, pages 101 / 214 91 / 195 of the light and heavy chains or alter (e.g., increase or decrease) the stability of the antibody.

[0170] In a particular embodiment, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant domain or FcRn binding fragment thereof (preferably an Fc domain fragment or Fc hinge) to alter (e.g., decrease or increase) the antibody half-life in vivo. See, for example, International Publications in WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631; and US Patents Nos. 5,869,046, 6,121,022, 6,277,375, and 6,165,745, all of which are incorporated herein by reference in their entirety, for examples of mutations that will alter (e.g., decrease or increase) the half-life of an antibody in vivo.In some embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant domain or FcRn-binding fragment thereof (preferably an Fc domain fragment or Fc-hinge) to alter and decrease the antibody's half-life in vivo. In other embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into an IgG constant domain or FcRn-binding fragment thereof (preferably an Fc domain fragment or Fc-hinge) to alter and increase the antibody's half-life in vivo. In a specific embodiment, antibodies may have one or more amino acid mutations (e.g., substitutions) in the second constant domain (CH2) (residues 231 to 340 of human IgG1) and / or the third constant domain (CH3) (residues 341 to 447 of human IgG1), numbered according to the EU numbering system.In a specific embodiment, the constant region of IgG1 of an antibody described herein comprises a methionine (M) to tyrosine (Y) substitution at position 252, a serine (S) to threonine (T) substitution at position 254, and a threonine (T) to glutamic acid (E) substitution at position 256, numbered from. Petition 870260065483, dated 03 / 07 / 2026, pages 102 / 214 92 / 195 according to the EU numbering system. See US Patent No. 7,658,921, which is incorporated herein by reference in its entirety. This type of mutant IgG, termed “YTE mutant,” has been shown to exhibit a fourfold increased half-life compared with wild-type versions of the same antibody (see Dall'Acqua WF et al., (2006) J Biol Chem 281: 23514-23524, which is incorporated herein by reference in its entirety). In certain embodiments, an antibody comprises a constant IgG domain comprising one, two, three, or more amino acid substitutions of amino acid residues at positions 251 to 257, 285 to 290, 308 to 314, 385 to 389, and 428 to 436, numbered according to the EU numbering system.

[0171] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of an antibody described herein (e.g., CH2 domain (residues 231 to 340 of human IgG1) and / or CH3 domain (residues 341 to 447 of human IgG1) and / or the hinge region, numbered according to the EU numbering system), to increase or decrease the antibody's affinity for an Fc receptor (e.g., an activated Fc receptor) on the surface of an effector cell. Mutations in the Fc region of an antibody that decrease or increase the antibody's affinity for an Fc receptor and techniques for introducing such mutations into the Fc receptor or a fragment thereof are known to one skilled in the art. Examples of mutations in the Fc receptor of an antibody that can be performed to alter the antibody's affinity for an Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109: 6181-6186, in Patent U.S. no. 6,737,056 and in International Publications in WO 02 / 060919; WO 98 / 23289; and WO 97 / 34631, all of which are incorporated herein by reference in their entirety.

[0172] In another embodiment, one, two or more amino acid substitutions are introduced into an Fc region of the IgG constant domain to alter the Petition 870260065483, dated 03 / 07 / 2026, pp. 103 / 214 93 / 195 effector function(s) of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, and 322, numbered according to the EU numbering system, may be substituted for a different amino acid residue so that the antibody has an altered affinity for an effector ligand but retains the antigen-binding capability of the original antibody. The effector ligand to which the affinity is altered may be, for example, an Fc receptor or the C1 component of complement. This approach is described in more detail in U.S. Patents Nos. 5,624,821 and 5,648,260, each of which is incorporated herein by reference in its entirety. In some modalities, deletion or inactivation (through point mutations or other means) of a constant region domain can reduce Fc receptor binding of circulating antibody, thereby increasing tumor localization.For example, see U.S. Patents Nos. 5,585,097 and 8,591,886, each of which is incorporated herein by reference in its entirety, for a description of mutations that delete or inactivate the constant domain and thus increase tumor localization. In certain embodiments, one or more amino acid substitutions may be introduced into the Fc region of an antibody described herein to remove potential glycosylation sites in the Fc region, which may reduce Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem 276: 6591-6604, which is incorporated herein by reference in its entirety).In several embodiments, one or more of the following mutations in the constant region of an antibody described herein may be performed: an N297A substitution; an N297Q substitution; an L235A substitution and an L237A substitution; an L234A substitution and an L235A substitution; an E233P substitution; an L234V substitution; an L235A substitution; a C236 deletion; a P238A substitution; a D265A substitution; an A327Q substitution; or a P329A substitution. Petition 870260065483, dated 03 / 07 / 2026, pages 104 / 214 94 / 195 numbered according to the EU numbering system. In certain embodiments, a mutation selected from the group consisting of D265A, P329A, and a combination thereof, numbered according to the EU numbering system, may be carried in the constant region of an antibody described in this document.

[0173] In one specific embodiment, an antibody described herein comprises the constant domain of an IgG1 with an amino acid substitution N297Q or N297A, numbered according to the EU numbering system. In another embodiment, an antibody described herein comprises the constant domain of an IgG1 with a mutation selected from the group consisting of D265A, P329A, and a combination thereof, numbered according to the EU numbering system. In another embodiment, an antibody described herein comprises the constant domain of an IgG1 with a mutation selected from the group consisting of L234A, L235A, and a combination thereof, numbered according to the EU numbering system.In certain embodiments, the amino acid residues in the constant region of an antibody described in this document at positions corresponding to L234, L235, and D265 in a human IgG1 heavy chain, numbered according to the EU numbering system, are not L, L, and D, respectively. This approach is described in detail in International Publication WO 14 / 108483, which is incorporated herein by reference in its entirety. In a particular embodiment, the amino acids corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain are F, E, and A; or A, A, and A, respectively, numbered according to the EU numbering system.

[0174] In certain embodiments, one or more amino acids selected from amino acid residues 329, 331 and 322 in the constant region of an antibody described herein, numbered according to the EU numbering system, Petition 870260065483, dated 03 / 07 / 2026, pp. 105 / 214 95 / 195 can be replaced by a different amino acid residue so that the antibody has altered C1q binding and / or reduced or abolished complement-dependent cytotoxicity (CDC). This approach is described in more detail in US Patent No. 6,194,551 (Idusogie et al.), which is incorporated herein by reference in its entirety. In some embodiments, one or more amino acid residues within amino acid positions 231 to 238 in the N-terminal region of the CH2 domain of an antibody described herein are altered to thereby change the antibody's ability to fix complement, numbered according to the EU numbering system. This approach is further described in International Publication WO 94 / 29351, which is incorporated herein by reference in its entirety.In certain embodiments, the Fc region of an antibody described in this document is modified to increase the antibody's ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or increase the antibody's affinity for an Fcy receptor by mutating one or more amino acids (e.g., introducing amino acid substitutions) at the following positions: 238, 239, 248, 249, 252, 254, 255, 256, 258, 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, 328, 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, numbered according to the EU numbering system. This approach is further described in International Publication WO 00 / 42072, which is incorporated herein by reference in its entirety.

[0175] In certain embodiments, an antibody described in this document comprises the constant region of an IgG4 antibody and the serine at amino acid residue 228 of the heavy chain, numbered according to the EU numbering system, is replaced by proline. In certain embodiments, the present invention provides Petition 870260065483, dated 03 / 07 / 2026, pp. 106 / 214 96 / 195 an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 74. In certain embodiments, the present invention provides an isolated antibody that specifically binds to TIM3 (e.g., human TIM-3), wherein the antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 75.

[0176] In certain embodiments, any of the constant region mutations or modifications described herein may be introduced into one or both of the heavy chain constant regions of an antibody described herein that has two heavy chain constant regions.

[0177] In certain embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3) and functions as an antagonist.

[0178] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and decreases the activity of TIM-3 (e.g., human TIM-3) by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% as assessed by methods described herein and / or known to one skilled in the art, relative to the activity of TIM-3 (e.g., human TIM-3) without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)).In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and decreases the activity of TIM-3 (e.g., human TIM-3) by at least about 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold. Petition 870260065483, dated 03 / 07 / 2026, pages 107 / 214 97 / 195 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times or 100 times as assessed by methods described herein and / or known to one skilled in the art, with respect to the activity of TIM-3 (e.g., human TIM-3) without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)). Non-limiting examples of TIM-3 (e.g., human TIM-3) activity may include TIM-3 (e.g., human TIM-3) signaling, binding of TIM-3 (e.g., human TIM-3) to TIM-3 (e.g., human TIM-3) ligand (e.g., phosphatidylserine), and inhibition of cytokine production (e.g., IFN-γ and / or TNFα). In certain embodiments, the present disclosure provides a single antibody that specifically binds to TIM-3 (e.g., human TIM-3) and disables, reduces, or inhibits TIM-3 activity (e.g., human TIM-3).In specific modalities, a decrease in TIM-3 activity (e.g., human TIM-3) is evaluated as described in the Examples below.

[0179] In specific embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and reduces the binding of TIM-3 (e.g., human TIM-3) to its ligand (e.g., phosphatidylserine) by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%, as assessed by the methods described herein (see Examples, below) or known to one skilled in the art, with respect to the binding of TIM-3 (e.g., human TIM-3) to its ligand (e.g., phosphatidylserine) without any antibody or with an unrelated antibody (for example, an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)).In specific embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and reduces the binding of TIM-3 (e.g., human TIM-3) to its ligand (e.g., phosphatidylserine) by at least approximately... Petition 870260065483, dated 03 / 07 / 2026, pp. 108 / 214 98 / 195 1.2 times, 1.3 times, 1.4 times, 1.5 times, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, 4.5 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times or 100 times, as assessed by the methods described herein (see Examples, below) or known to one skilled in the art, with respect to the binding of TIM-3 (e.g., human TIM-3) to its ligand (e.g., phosphatidylserine) without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)).

[0180] In specific embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and increases cytokine production (e.g., IFN-γ and / or TNF-α) by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%, as assessed by the methods described in this document (see Examples below) or known to one skilled in the art, relative to cytokine production without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)).In specific embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM3 (e.g., human TIM-3) and increases cytokine production (e.g., IFNγ and / or TNF-α) by at least about 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold, as assessed by methods described herein (see Examples, below) or known to one skilled in the art, relative to cytokine production without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM3). TIM-3 (e.g., human TIM-3). Petition 870260065483, dated 03 / 07 / 2026, pp. 109 / 214 99 / 195

[0181] In specific embodiments, the present disclosure provides a single antibody that specifically binds to TIM-3 (e.g., human TIM-3) and, alone or in combination with an anti-PD-1 antibody (e.g., pembrolizumab or nivolumab), increases IFN-γ production in human peripheral blood mononuclear cells (PBMCs) in response to Staphylococcus Enterotoxin A (SEA) stimulation by at least about 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold, as appropriate. evaluated by methods described in this document (see Examples below) or known to one skilled in the art, regarding the production of IFN-γ without any antibody or with an unrelated antibody (e.g., an antibody that does not specifically bind to TIM-3 (e.g., human TIM-3)).

[0182] In certain embodiments, human peripheral blood mononuclear cells (PBMCs) stimulated with Staphylococcus A Enterotoxin A (SEA) in the presence of an antibody described in this document that specifically binds to TIM-3 (e.g., human TIM-3) have increased IFN-γ production by at least approximately 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold compared to PBMCs stimulated only with SEA without any antibody or with an unrelated antibody (e.g., an antibody that does not bind). specifically TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art.

[0183] In specific embodiments, the present disclosure provides an isolated antibody that binds specifically to TIM-3 (e.g., human TIM-3) and Petition 870260065483, dated 03 / 07 / 2026, pp. 110 / 214 100 / 195 alone or in combination with an anti-PD-1 antibody (e.g., pembrolizumab or nivolumab) increases the production of IFN-γ and / or TNFα in tumor-infiltrating lymphocytes (TILs) in response to anti-CD3 antibody and anti-CD28 antibody stimulation by at least about 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold, as assessed by methods described in this document (see Examples below) or known to one skilled in the art, with respect to the production of IFN-γ and / or TNFα without an antibody that specifically binds to TIM-3 (e.g., human TIM-3). In one embodiment, the TILs are from non-small cell lung cancer (NSCLC) tumors. In another embodiment, the TILs are from gallbladder adenocarcinoma tumors.In another scenario, the TILs are related to breast cancer tumors.

[0184] In certain embodiments, tumor-infiltrating lymphocytes (TILs) stimulated with anti-CD3 and anti-CD28 antibodies in the presence of an antibody described in this document that specifically binds to TIM-3 (e.g., human TIM-3) have increased IFN-γ and / or TNFα production by at least approximately 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold compared to TILs stimulated only with anti-CD3 and anti-CD28 antibodies without an antibody that specifically binds to TIM-3. TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art. In one embodiment, the TILs are from non-small cell lung cancer (NSCLC) tumors.In another instance, the TILs are from gallbladder adenocarcinoma tumors. In yet another instance, the TILs are from breast cancer tumors. Petition 870260065483, dated 03 / 07 / 2026, pages 111 / 214 101 / 195

[0185] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3) and is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3). In the specific embodiments, at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% of the antibody described herein is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described herein (see Examples, below) or known to one skilled in the art. In certain embodiments, a lower percentage of cells expressing TIM-3 (e.g., human TIM-3) survive in the presence of the antibody described herein than in the presence of a reference anti-TIM-3 antibody (e.g., human TIM-3) in an assay comprising the following steps: (a) Place the cells in plates expressing TIM-3 (e.g., human TIM-3) at 2 x 104 cells per well in a tissue culture plate; (b) add the same concentrations of αHFc-NC-DM1 and the antibody described herein or the reference anti-TIM-3 antibody (e.g., human TIM-3) (e.g., 1.5 ng / ml, 4.6 ng / ml, 13.7 ng / ml, 41.2 ng / ml, 123.5 ng / ml, 370 ng / ml, 1111 ng / ml or 3333 ng / ml) to a final volume of 100 μl / well; (c) incubate at 37°C and 5% CO2 for 72 hours; (d) measure the survival of cells expressing TIM-3 (e.g., human TIM-3); and (e) calculate the percentage of cell survival relative to untreated cells expressing TIM-3 (e.g., human TIM-3).

[0186] In certain embodiments, the percentage of cell survival in the presence of the antibody described herein is at least approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, Petition 870260065483, dated 03 / 07 / 2026, pages 112 / 214 102 / 195 85%, 90%, 95%, 98%, or 99% lower than the percentage of cell survival in the presence of the reference anti-TIM-3 antibody (e.g., human TIM-3). In certain embodiments, the percentage of cell survival in the presence of the antibody described herein is at least approximately 1.2 times, 1.3 times, 1.4 times, 1.5 times, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, 4.5 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times, or 100 times lower than the percentage of cell survival in the presence of the reference anti-TIM-3 antibody (e.g., human TIM-3). In certain embodiments, the reference anti-TIM-3 antibody (e.g., human TIM-3) is pab1944w (IgG1 N297A). In certain embodiments, the reference anti-TIM-3 antibody (e.g., human TIM-3) is Hum11 (IgG4 S228P).In certain embodiments, the cells expressing TIM-3 (e.g., human TIM-3) are Kasumi-3 cells. In certain embodiments, the cells expressing TIM-3 (e.g., human TIM-3) are Kasumi-3 cells (ATCC® CRL-2725™). In certain embodiments, the cells expressing TIM-3 (e.g., human TIM-3) are Jurkat cells genetically modified to express TIM-3 (e.g., human TIM-3).

[0187] In certain embodiments, up to 50% of cells expressing TIM-3 (e.g., human TIM-3) survive in the presence of the antibody described herein compared to untreated cells expressing TIM-3 (e.g., human TIM-3) in an assay comprising the following steps: (a) Place the cells in plates expressing TIM-3 (e.g., human TIM-3) at 2 x 104 cells per well in a tissue culture plate; (b) add the same concentrations of αHFc-NC-DM1 and the antibody described in this document (e.g., 1.5 ng / ml, 4.6 ng / ml, 13.7 ng / ml, 41.2 ng / ml, 123.5 ng / ml, 370 ng / ml, 1111 ng / ml or 3333 ng / ml) to a final volume of 100 μl / well; Petition 870260065483, dated 03 / 07 / 2026, pages 113 / 214 103 / 195 (c) incubate at 37°C and 5% CO2 for 72 hours; (d) measure the survival of cells expressing TIM-3 (e.g., human TIM-3); and (e) calculate the percentage of cell survival relative to untreated cells expressing TIM-3 (e.g., human TIM-3).

[0188] In certain embodiments, at most 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of cells expressing TIM-3 (e.g., human TIM-3) survive in the presence of the antibody described herein compared to untreated cells expressing TIM-3 (e.g., human TIM-3). In certain embodiments, αHFc-NC-DM1 and the antibody described herein are added at a concentration of 1111 ng / ml, and a maximum of 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of cells expressing TIM-3 (e.g., human TIM-3) survive in the presence of the antibody described herein compared to untreated cells expressing TIM-3 (e.g., human TIM-3).In certain embodiments, αHFc-NC-DM1 and the antibody described herein are added at a concentration of 1111 ng / ml, and a maximum of 50% of cells expressing TIM-3 (e.g., human TIM-3) survive in the presence of the antibody described herein compared to untreated cells expressing TIM-3 (e.g., human TIM-3).

[0189] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described herein (see Examples, below) or known to one skilled in the art, and wherein the antibody comprises a CDRH1 comprising the amino acid sequence X1X2X3X4X5S Petition 870260065483, dated 03 / 07 / 2026, pp. 114 / 214 104 / 195 (SEQ ID NO: 48), in which Xi is R, S, A, G, K, M, or T. X2 is Q, S, A, G, R, or T. X3 is N, Y, G, or Q. X4 is A or Q, and X5 is W, M, A, S, or T.

[0190] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRH1 comprising the amino acid sequence X1X2NAWS (SEQ ID NO: 49), wherein X1 is either R or A; and X2 is either Q or R.

[0191] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRH1 comprising the amino acid sequence X1X2GQX3S (SEQ ID NO: 50), wherein X1 is K, M, or G; Petition 870260065483, dated 03 / 07 / 2026, pp. 115 / 214 105 / 195 X2 is A or S; and X3 is either S or T.

[0192] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRH1 comprising the amino acid sequence X1X2QQAS (SEQ ID NO: 51), wherein X1 is S, R, T, or G; and X2 is A, S, T, or G.

[0193] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples, below) or known to one skilled in the art, and wherein the antibody comprises a CDRH2 comprising the amino acid sequence WVSAISGSGGSTY (SEQ ID NO: 2).

[0194] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), Petition 870260065483, dated 03 / 07 / 2026, pp. 116 / 214 106 / 195 as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRH3 comprising the amino acid sequence AKGGDYGGNYFD (SEQ ID NO: 3).

[0195] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRL1 comprising the amino acid sequence X1ASQSVX2SSYLA (SEQ ID NO: 52), wherein X1 is R or G; and X2 is missing or it is S.

[0196] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples, below) or known to one skilled in the art, and wherein the antibody comprises a CDRL2 comprising the amino acid sequence X1ASX2RAT ​​(SEQ ID NO: 53), wherein X1 is D or G; and X2 is N, S, or T.

[0197] In certain embodiments, the present disclosure provides an antibody Petition 870260065483, dated 03 / 07 / 2026, pp. 117 / 214 107 / 195 isolate that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a CDRL3 comprising the amino acid sequence QQYGSSPXiT (SEQ ID NO: 54), wherein Xi is L or I.

[0198] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody cross-competes for binding to TIM-3 (e.g., human TIM-3) with an antibody comprising the variable region heavy and light chain amino acid sequences presented in the SEQ IDs. Nos: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 27 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46;33 and 46; 34 and 46; or 35 and 46, respectively.

[0199] In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), Petition 870260065483, dated 03 / 07 / 2026, pp. 118 / 214 108 / 195 as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody binds to the same epitope or an overlapping epitope of TIM-3 (e.g., a human TIM3 epitope) as an antibody described in this document, for example, an antibody comprising the variable region heavy and light chain amino acid sequences presented in the SEQ ID NOs: 24 and 36; 24 and 38; 26 and 42; 24 and 42; 24 and 46; 24 and 43; 26 and 43; 26 and 46; 26 and 41; 24 and 41; 25 and 39; 24 and 47; 25 and 40; 26 and 47; 25 and 37; 25 and 45; 25 and 44; 25 and 46; 25 and 42; 25 and 41; 25 and 43; 25 and 47; 25 and 46; 28 and 46; 29 and 46; 30 and 46; 31 and 46; 32 and 46; 33 and 46; 34 and 46; or 35 and 46, respectively.In certain embodiments, the present disclosure provides an isolated antibody that specifically binds to TIM-3 (e.g., human TIM-3), wherein at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% of the antibody is internalized after binding to cells expressing TIM-3 (e.g., human TIM-3), as assessed by methods described in this document (see Examples below) or known to one skilled in the art, and wherein the antibody comprises a human IgG heavy chain constant region that is a variant of a wild-type human IgG heavy chain constant region, wherein the variant human IgG heavy chain constant region binds to an Fc gamma receptor. Human IgG with lower affinity than the wild-type human heavy chain constant region binds to the human Fc gamma receptor.In certain embodiments, the human Fc gamma receptor is selected from the group consisting of FcyRI, FcyRII, and FcyRIII. In certain embodiments, the variant human IgG heavy chain constant region is an IgG1 constant region comprising an N297A mutation, numbered according to the EU numbering system. 6.3 Pharmaceutical Composition Petition 870260065483, dated 03 / 07 / 2026, pp. 119 / 214 109 / 195

[0200] Compositions comprising an anti-TIM-3 antibody (e.g., human TIM-3) described herein having the desired degree of purity in a physiologically acceptable carrier, excipient, or stabilizer are provided herein (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are non-toxic to receptors at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenolic, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight polypeptides (less than about 10 residues);Proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0201] In a specific embodiment, the pharmaceutical compositions comprise an anti-TIM-3 antibody (e.g., human TIM-3) described in this document and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable vehicle. In a specific embodiment, the pharmaceutical compositions comprise an effective amount of an antibody described in this document and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable vehicle. In some embodiments, the antibody is the only active ingredient included in the pharmaceutical composition. Petition 870260065483, dated 03 / 07 / 2026, pp. 120 / 214 110 / 195 The pharmaceutical compositions described in this document may be useful for inhibiting the activity of TIM-3 (e.g., human TIM-3) and treating a condition, such as cancer or an infectious disease. In one embodiment, the present invention relates to a pharmaceutical composition of the present invention comprising an anti-TIM-3 antibody of the present invention for use as a medicament. In another embodiment, the present invention relates to a pharmaceutical composition of the present invention for use in a method for treating cancer or an infectious disease. In yet another embodiment, the present invention relates to the use of a pharmaceutical composition of the invention to prepare a medicament for treating cancer or an infectious disease.

[0202] Pharmaceutically acceptable vehicles used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include Sodium Chloride Injection, Ringers Injection, Isotonic Dextrose Injection, Sterile Water Injection, Dextrose, and Lactated Ringers Injection. Non-aqueous parenteral vehicles include fixed vegetable oils, cottonseed oil, corn oil, sesame oil, and peanut oil.Antimicrobial agents in bacteriostatic or fungistatic concentrations may be added to parenteral preparations packaged in multidose containers, including phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include carboxymethylcellulose. Petition 870260065483, dated 03 / 07 / 2026, page 121 / 214. 111 / 195 dica, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Emulsifying agents include polysorbate 80 (TWEEN® 80). A metal ion chelating or sequestering agent includes EDTA. Pharmaceutical vehicles also include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0203] A pharmaceutical composition may be formulated for any route of administration to an individual. Specific examples of routes of administration include intranasal, oral, pulmonary, transdermal, intradermal, and parenteral. Parenteral administration, characterized by subcutaneous, intramuscular, or intravenous injection, is also contemplated in this document. Injectables may be prepared in conventional forms, such as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline solution, dextrose, glycerol, or ethanol.Additionally, if desired, the pharmaceutical compositions to be administered may also contain smaller amounts of non-toxic auxiliary substances, such as humectants and emulsifiers, pH buffering agents, stabilizers, solubility enhancers and other similar agents, such as, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins.

[0204] Preparations for parenteral administration of an antibody include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent shortly before use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle shortly before use, and sterile emulsions. Solutions may be aqueous or non-aqueous.

[0205] If administered intravenously, suitable vehicles include Petition 870260065483, dated 03 / 07 / 2026, pages 122 / 214 112 / 195 physiological saline solution or phosphate-buffered saline (PBS) and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol and polypropylene glycol and mixtures thereof.

[0206] Topical mixtures comprising an antibody are prepared as described for local and systemic administration. The resulting mixture may be a solution, suspension, emulsion or similar and may be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal plasters or any other formulations for topical administration.

[0207] An anti-TIM-3 antibody (e.g., human TIM-3) described herein may be formulated as an aerosol for topical application, such as by inhalation (see, for example, U.S. Patents Nos. 4,044,126, 4,414,209 and 4,364,923, which describe aerosols for delivering a steroid useful for the treatment of inflammatory diseases, particularly asthma, and are incorporated herein by reference in their entirety). Such formulations for administration to the respiratory tract may be in the form of an aerosol or solution for a nebulizer or as a microfine powder, alone or in combination with an inert carrier, such as lactose. In such case, the particles of the formulation will, in one embodiment, have diameters smaller than 50 microns, in another embodiment, smaller than 10 microns.

[0208] An anti-TIM-3 antibody (e.g., human TIM-3) described herein may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eyes, in the form of gels, creams and lotions, and for application to the eyes or for intracisternal or intraspinal application. Topical administration is contemplated for transdermal distribution and also for administration to the eyes or mucous membranes or for inhalation therapies. Then Petition 870260065483, dated 03 / 07 / 2026, pp. 123 / 214 113 / 195 Nasal solutions of the antibody alone or in combination with other pharmaceutically acceptable excipients may also be administered.

[0209] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those skilled in the art and can be used to administer an antibody. For example, such patches are disclosed in U.S. Patents Nos. 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010,715, 5,985,317, 5,983,134, 5,948,433 and 5,860,957, all of which are incorporated herein by reference in their entirety.

[0210] In certain embodiments, a pharmaceutical composition comprising an antibody described herein is a lyophilized powder, which can be reconstituted for administration as solutions, emulsions, and other mixtures. It can also be reconstituted and formulated as solids or gels. The lyophilized powder is prepared by dissolving an antibody described herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. In some embodiments, the lyophilized powder is sterile. The solvent may contain an excipient that enhances the stability or another pharmacological component of the powder or reconstituted solution prepared from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agent.The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate, or another similar buffer known to those skilled in the art, in one embodiment, at a pH close to neutral. Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those skilled in the art provides the desired formulation. In one embodiment, the resulting solution will be divided into vials for lyophilization. Each vial will contain a single dosage or multiple dosages of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4°C. Petition 870260065483, dated 03 / 07 / 2026, pp. 124 / 214 114 / 195 room temperature. Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or another suitable vehicle. The exact amount depends on the compound selected. This amount can be determined empirically.

[0211] The anti-TIM-3 antibodies (e.g., TIM-3) described in this document and other compositions provided herein may also be formulated to be targeted to a particular tissue, receptor, or other area of ​​the body of the individual to be treated. Many such targeting methods are well known to those skilled in the art. All such targeting methods are contemplated herein for use in the present compositions. For non-limiting examples of bleaching methods, see, for example, U.S. Patents Nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542 and 5,709,874, all of which are incorporated herein by reference in their entirety. In one specific embodiment, an antibody described in this document targets a tumor.

[0212] The compositions to be used for in vivo administration may be sterile. This is readily achieved by filtration through, for example, sterile filtration membranes. 6.4 METHODS OF USE AND USES

[0213] In another aspect, the present disclosure provides a method for treating an individual using the anti-TIM-3 antibodies (e.g., human TIM-3) disclosed herein. Any disease or disorder in an individual that could benefit from inhibition of TIM-3 function (e.g., human TIM-3) can be treated using the anti-TIM-3 antibodies (e.g., human TIM-3). Petition 870260065483, dated 03 / 07 / 2026, pp. 125 / 214 115 / 195 disclosed in this document. The anti-TIM-3 antibodies (e.g., human TIM-3) disclosed in this document are particularly useful for inhibiting immune system tolerance to tumors and, consequently, can be used as immunotherapy for individuals with cancer. For example, in certain embodiments, this disclosure provides a method for increasing T cell activation in response to an antigen in an individual, wherein the method comprises administering to the individual an effective amount of an anti-TIM-3 antibody (e.g., human TIM-3) or pharmaceutical composition thereof, as disclosed in this document. In certain embodiments, this disclosure provides a method for treating cancer in an individual, wherein the method comprises administering to the individual an effective amount of the antibody or pharmaceutical composition as disclosed in this document.In certain embodiments, this disclosure provides an antibody or pharmaceutical composition as disclosed herein for use in a method for treating cancer or an infectious disease. In certain embodiments, this disclosure provides an antibody or pharmaceutical composition as disclosed herein for use as a medicament. In another embodiment, this disclosure provides the use of an antibody or pharmaceutical composition as disclosed herein to prepare a medicament for treating cancer or an infectious disease.

[0214] Cancers that can be treated with anti-TIM-3 antibodies (e.g., human TIM-3) or pharmaceutical compositions disclosed herein include, without limitation, a solid tumor, a hematologic cancer (e.g., leukemia, lymphoma, myeloma, e.g., multiple myeloma), and a metastatic lesion. In one embodiment, the cancer is a solid tumor. Examples of solid tumors include malignancies, e.g., sarcomas and carcinomas, e.g., adenocarcinomas of various organ systems, such as those affecting the lung, Petition 870260065483, dated 03 / 07 / 2026, pages 126 / 214 116 / 195 breast, ovarian, lymphoid, gastrointestinal (e.g., colon), anal, genital and gastrointestinal tract (e.g., renal, urothelial, bladder cells, prostate), pharynx, CNS (e.g., brain, neural or glial cells), head and neck, skin (e.g., melanoma) and pancreas, as well as adenocarcinomas which include malignancies such as colon cancer, rectal cancer, renal cell carcinoma, liver cancer, lung cancer (e.g., non-small cell lung cancer or small cell lung cancer), small bowel cancer and esophageal cancer. The cancer may be early, intermediate, late stage or metastatic. In certain modalities, the cancer is associated with elevated PD-1 activity (e.g., elevated PD-1 expression).

[0215] In one embodiment, the cancer is chosen from among a lung cancer (e.g., lung adenocarcinoma or non-small cell lung cancer (NSCLC) (e.g., an NSCLC with squamous and / or non-squamous histology or an NSCLC adenocarcinoma)), a melanoma (e.g., an advanced melanoma), a renal cancer (e.g., a renal cell carcinoma), a liver cancer (e.g., hepatocellular carcinoma), a myeloma (e.g., multiple myeloma), a prostate cancer, a breast cancer (e.g., a breast cancer that does not express one, two, or all of the estrogen receptor, progesterone receptor, or Her2 / neu, e.g., a triple-negative breast cancer), an ovarian cancer, a colorectal cancer, a pancreatic cancer, a head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC)), anal cancer, gastroesophageal cancer (e.g., squamous cell carcinoma esophageal), mesothelioma, nasopharyngeal cancer,Thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, or a lymphoproliferative disease (e.g., post-transplant lymphoproliferative disease). In one embodiment, the cancer is NSCLC. In one embodiment, the cancer is renal cell carcinoma. In one embodiment, the cancer is ovarian cancer. In a specific embodiment, the ovarian cancer is a... Petition 870260065483, dated 03 / 07 / 2026, pages 127 / 214 117 / 195 platinum-refractive ovarian cancer.

[0216] In one form, the cancer is a hematological cancer, for example, leukemia, lymphoma, or myeloma. In another form, the cancer is a leukemia, for example, acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), acute myeloblastic leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), chronic lymphocytic leukemia (CLL), or hairy cell leukemia.In one modality, the cancer is a lymphoma, for example, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like diffuse large B-cell lymphoma (ABC), diffuse large central germ cell (GCB) lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, recurrent follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, or extranodal marginal zone lymphoma. In another modality, the cancer is a myeloma, for example, multiple myeloma.

[0217] In another modality, the cancer is chosen from among a carcinoma (for example, advanced or metastatic carcinoma), melanoma, or a lung carcinoma, for example, a non-small cell lung carcinoma.

[0218] In one modality, the cancer is lung cancer, for example, lung adenocarcinoma, non-small cell lung cancer, or small cell lung cancer.

[0219] In one embodiment, the cancer is a melanoma, for example, an advanced melanoma. In another embodiment, the cancer is an advanced or unresectable melanoma that does not respond to other therapies. In other embodiments, the cancer is a melanoma with a BRAF mutation (for example, a BRAF V600 mutation). In still other embodiments, the anti-TIM-3 antibody (for example, human TIM-3) or pharmaceutical composition disclosed herein is administered after Petition 870260065483, dated 03 / 07 / 2026, pages 128 / 214 118 / 195 treatment with an anti-CTLA-4 antibody (e.g., ipilimumab) with or without a BRAF inhibitor (e.g., vemurafenib or dabrafenib).

[0220] In another form, the cancer is a hepatocellular carcinoma, for example, an advanced hepatocellular carcinoma, with or without a viral infection, for example, chronic viral hepatitis.

[0221] In another modality, the cancer is prostate cancer, for example, advanced prostate cancer.

[0222] In yet another form, the cancer is a myeloma, for example, multiple myeloma.

[0223] In yet another modality, the cancer is a renal cancer, for example, a renal cell carcinoma (RCC) (e.g., a metastatic RCC, clear cell renal cell carcinoma (CRCCC) or renal papillary cell carcinoma).

[0224] In yet another modality, the cancer is chosen from among lung cancer, melanoma, kidney cancer, breast cancer, colorectal cancer, leukemia, or a metastatic cancer lesion.

[0225] In certain embodiments, this disclosure provides a method for preventing or treating an infectious disease in an individual, wherein the method comprises administering to the individual an effective amount of an anti-TIM-3 antibody (e.g., human TIM-3) or a pharmaceutical composition thereof, as disclosed in this document. In one embodiment, methods for preventing and / or treating an infection (e.g., a viral infection, a bacterial infection, a fungal infection, a protozoan infection, or a parasitic infection) are provided in this document. The infection prevented and / or treated in accordance with the methods may be caused by an infectious agent identified in this document. In a specific embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) described in this document or a composition thereof is the only active agent administered to an individual. In some embodiments, an anti-TIM-3 antibody (e.g., human TIM-3) Petition 870260065483, dated 03 / 07 / 2026, pp. 129 / 214 119 / 195 example, human TIM-3) described in this document or a composition thereof is used in combination with anti-infective interventions (e.g., antivirals, antibacterials, antifungals or anthelmintics) for the treatment of infectious diseases. Therefore, in one embodiment, the present invention relates to an antibody and / or pharmaceutical composition of the present invention for use in a method for preventing and / or treating an infectious disease, optionally wherein the antibody or pharmaceutical composition is the only active agent administered to an individual or wherein the antibody or pharmaceutical composition is used in combination with anti-infective interventions.

[0226] The infectious diseases that can be treated and / or prevented by anti-TIM-3 antibodies (e.g., human TIM-3) or pharmaceutical compositions disclosed herein are caused by infectious agents, including, but not limited to, bacteria, parasites, fungi, protozoa, and viruses. In one specific embodiment, the infectious disease treated and / or prevented by anti-TIM3 antibodies (e.g., human TIM-3) or pharmaceutical compositions disclosed herein is caused by a virus.Viral diseases or viral infections that can be prevented and / or treated in accordance with the methods described in this document include, but are not limited to, those caused by hepatitis A, hepatitis B, hepatitis C, influenza (e.g., influenza A or influenza B), varicella, adenovirus, herpes simplex type I (HSV-I), herpes simplex type II (HSV-II), rinderpest, rhinovirus, echovirus, rotavirus, respiratory syncytial virus, papillomavirus, papova virus, cytomegalovirus, echinovirus, arbovirus, huntavirus, coxsackievirus, mumps virus, measles virus, rubella virus, poliovirus, smallpox, Epstein-Barr virus, human immunodeficiency virus type I (HIV-I), human immunodeficiency virus type II (HIV-II), and viral disease agents such as meningitis, encephalitis, dengue fever, or smallpox. Petition 870260065483, dated 03 / 07 / 2026, pages 130 / 214 120 / 195

[0227] Bacterial infections that can be prevented and / or treated include infections caused by Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Proteus vulgaris, Staphylococcus viridans, and Pseudomonas aeruginosa. Bacterial diseases caused by bacteria (e.g., Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Proteus vulgaris, Staphylococcus viridans, and Pseudomonas aeruginosa) that can be prevented and / or treated in accordance with the methods described herein include, but are not limited to, Mycobacteria rickettsia, Mycoplasma, Neisseria, S. pneumoniae, Borrelia burgdorferi (Lyme disease), Bacillus anthracis (anthrax), tetanus, Streptococcus, Staphylococcus, mycobacteria, pertussis, cholera, plague, diphtheria, chlamydia, S. aureus, and Legionella.

[0228] Protozoan diseases or protozoan infections caused by protozoa that can be prevented and / or treated in accordance with the methods described in this document include, but are not limited to, leishmaniasis, coccidiosis, trypanosomiasis, schistosomiasis, or malaria. Parasitic diseases or parasitic infections caused by parasites that can be prevented and / or treated in accordance with the methods described in this document include, but are not limited to, chlamydia and rickettsia.

[0229] Fungal diseases or fungal infections that can be prevented and / or treated in accordance with the methods described in this document include, but are not limited to, those caused by Candida infections, zygomycosis, Candida mastitis, progressive disseminating trichosporonosis with latent trichosporonemia, disseminated candidiasis, pulmonary paracoccidioidomycosis, pulmonary aspergillosis, Pneumocystis carinii pneumonia, cryptococcal meningitis, coccidioidal meningoencephalitis and cerebrospinal vasculitis, Aspergillus niger infection, Fusarium keratitis, paranasal sinus mycoses, Aspergillus fumigatus endocarditis, discoloration Petition 870260065483, dated 03 / 07 / 2026, pages 131 / 214 121 / 195 tibial dysplasia, Candida glabrata vaginitis, oropharyngeal candidiasis, X-linked chronic granulomatous disease, athlete's foot, cutaneous candidiasis, fungal placentitis, disseminated trichosporonosis, allergic bronchopulmonary aspergillosis, fungal keratitis, Cryptococcus neoformans infection, fungal peritonitis, Curvularia geniculata infection, staphylococcal endophthalmitis, sporotrichosis, and dermatophytosis.

[0230] In certain embodiments, these methods additionally comprise administering an additional therapeutic agent to the individual. In certain embodiments, the additional therapeutic agent is a chemotherapeutic product, a radiotherapeutic product, or a checkpoint targeting agent. In certain embodiments, the chemotherapeutic agent is a hypomethylating agent (e.g., azacitidine). In certain embodiments, the checkpoint targeting agent is selected from the group consisting of an anti-CTLA-4 antagonist antibody, an anti-PD-L1 antagonist antibody, an anti-PD-L2 antagonist antibody, an anti-PD-1 antagonist antibody, an anti-TIM-3 antagonist antibody, an anti-LAG-3 antagonist antibody, an anti-CEACAM1 antagonist antibody, an anti-CD137 agonist antibody, an anti-TIGIT antagonist antibody, an anti-VISTA antagonist antibody, an anti-GITR antagonist antibody, and an anti-OX40 agonist antibody.

[0231] In one embodiment, the present invention relates to an antibody and / or pharmaceutical composition of the present invention for use in a method of the present invention, wherein the method further includes administering an additional therapeutic agent to the individual. In one embodiment, the present invention relates to (a) an antibody and / or pharmaceutical composition of the present invention and (b) an additional therapeutic agent for use as a medicament. In another embodiment, the present invention relates to (a) an antibody and / or pharmaceutical composition of the present invention and (b) an additional therapeutic agent for use in a method for the treatment of cancer. In another embodiment, the present invention relates to a pharmaceutical composition, kit or kit of parts comprising (a) an antibody and / or composition Petition 870260065483, dated 03 / 07 / 2026, pp. 132 / 214 122 / 195 pharmaceutical of the present invention and (b) an additional therapeutic agent. In one embodiment, the additional therapeutic agent is a chemotherapeutic product, a radiotherapeutic product, or a checkpoint bleaching agent.

[0232] In certain embodiments, an anti-PD-1 antibody is used in methods disclosed in this document. In certain embodiments, the anti-PD-1 antibody is nivolumab, also known as BMS-936558 or MDX1106, developed by Bristol-Myers Squibb. In certain embodiments, the anti-PD-1 antibody is pembrolizumab, also known as lambrolizumab or MK-3475, developed by Merck & Co. In certain embodiments, the anti-PD-1 antibody is pidilizumab, also known as CT-011, developed by CureTech. In certain embodiments, the anti-PD-1 antibody is MEDI0680, also known as AMP-514, developed by Medimmune. In certain embodiments, the anti-PD-1 antibody is PDR001 developed by Novartis Pharmaceuticals. In certain embodiments, the anti-PD-1 antibody is REGN2810 developed by Regeneron Pharmaceuticals. In certain embodiments, the anti-PD-1 antibody is PF-06801591 developed by Pfizer. In certain embodiments, the anti-PD-1 antibody is BGB-A317 developed by BeiGene.In certain embodiments, the anti-PD-1 antibody is TSR-042 developed by AnaptysBio and Tesaro. In certain embodiments, the anti-PD-1 antibody is SHR-1210 developed by Hengrui.

[0233] Other non-limiting examples of anti-PD-1 antibodies that may be used in treatment methods disclosed herein are disclosed in the following patents and patent applications, all of which are incorporated herein by reference in their entirety for all purposes: U.S. Patent No. 6,808,710; U.S. Patent No. 7,332,582; U.S. Patent No. 7,488,802; U.S. Patent No. 8,008,449; U.S. Patent No. Try US patent no. 8,354,509; US Patent Try US No. 8,747,847; US Patent no 8,114,845; US Patent No. 8,168,757; PA no. 8,686,119; US Patent No. 8,735,553; Pa no 8,779,105; US Patent No. 8,927,697; Shovel Petition 870260065483, dated 03 / 07 / 2026, pp. 133 / 214 123 / 195, US patent no. 8,993,731; US ​​patent no. 9,102,727; US patent no. 9,205,148; US publication no. 2013 / 0202623 A1; US ​​publication no. 2013 / 0291136 A1; US ​​publication no. 2014 / 0044738 A1; US ​​publication no. 2014 / 0356363 A1; US ​​publication no. 2016 / 0075783 A1; and PCT publication no. WO 2013 / 033091 A1; PCT publication no. WO 2015 / 036394 A1; PCT publication no. WO 2014 / 179664 A2; PCT publication no. WO 2014 / 209804 A1; PCT publication in WO 2014 / 206107 A1; PCT publication in WO 2015 / 058573 A1; PCT publication in WO 2015 / 085847 A1; PCT publication in WO 2015 / 200119 A1; PCT publication in WO 2016 / 015685 A1; PCT publication in WO TOC 2016 / 020856 A1.

[0234] In certain embodiments, an anti-PD-L1 antibody is used in methods disclosed in this document. In certain embodiments, the anti-PD-L1 antibody is atezolizumab developed by Genentech. In certain embodiments, the anti-PD-L1 antibody is durvalumab developed by AstraZeneca, Celgene, and Medimmune. In certain embodiments, the anti-PD-L1 antibody is avelumab, also known as MSB0010718C, developed by Merck Serono and Pfizer. In certain embodiments, the anti-PD-L1 antibody is MDX-1105 developed by Bristol-Myers Squibb. In certain embodiments, the anti-PD-L1 antibody is AMP-224 developed by Amplimmune and GSK.

[0235] Non-limiting examples of anti-PD-L1 antibodies that can be used in treatment methods disclosed herein are disclosed in the following patents and patent applications, all of which are incorporated herein by reference in their entirety for all purposes: U.S. Patent No. 7,943,743; U.S. Patent No. 8,168,179; U.S. Patent No. 8,217,149; U.S. Patent No. 8,552,154; U.S. Patent No. 8,779,108; U.S. Patent No. 8,981,063; U.S. Patent No. 9,175,082; U.S. Publication No. 2010 / 0203056 A1; U.S. Publication No. 2003 / 0232323 A1; PCT Publication No. WO 2013 / 0323249 A1; PCT publication in WO 2014 / 0341917 A1; PCT publication in WO 2014 / 0044738 A1; PCT publication in WO 2015 / 0203580 A1; Petition 870260065483, dated 03 / 07 / 2026, pages 134 / 214 124 / 195 PCT Publication in WO 2015 / 0225483 A1; PCT Publication in WO 2015 / 0346208 A1; US ​​Publication in TOC 2015 / 0355184 A1; and PCT Publication in WO 2014 / 100079 A1; PCT Publication in WO 2014 / 022758 A1; PCT Publication in WO 2014 / 055897 A2; PCT Publication in WO 2015 / 061668 A1; PCT Publication in WO 2015 / 109124 A1; PCT Publication in WO 2015 / 195163 A1; PCT Publication in WO 2016 / 000619 A1; and PCT Publication in WO 2016 / 030350 A1.

[0236] In certain embodiments, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed in this document is administered to an individual in combination with a compound that targets an immunomodulatory enzyme (or enzymes), such as IDO (indoleamine-(2,3)-dioxygenase) and / or TDO (tryptophan 2,3-dioxygenase). Therefore, in one embodiment, the additional therapeutic agent is a compound that targets an immunomodulatory enzyme (or enzymes), such as an indoleamine-(2,3)-dioxygenase (IDO) inhibitor. In certain embodiments, such a compound is selected from the group consisting of epacadostate (Incyte Corp; see, for example, WO 2010 / 005958, which is incorporated herein by reference in its entirety), F001287 (Flexus Biosciences / BristolMyers Squibb), indoximode (NewLink Genetics), and NLG919 (NewLink Genetics). In one embodiment, the compound is epacadostate. In another embodiment, the compound is F001287. In yet another embodiment, the compound is indoximode.In another embodiment, the compound is NLG919. In a specific embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed herein is administered to an individual in combination with an IDO inhibitor to treat cancer. The IDO inhibitor, as described herein for use in cancer treatment, is present in a solid dosage form of a pharmaceutical composition, such as a tablet, pill, or capsule, wherein the pharmaceutical composition includes an IDO inhibitor and a pharmaceutically acceptable excipient. As such, the antibody as described herein and the IDO inhibitor as described in... Petition 870260065483, dated 03 / 07 / 2026, pages 135 / 214 125 / 195 of this document may be administered separately, sequentially, or concomitantly as separate dosage forms. In one embodiment, the antibody is administered parenterally, and the IDO inhibitor is administered orally. In particular embodiments, the inhibitor is selected from the group consisting of epacadostat (Incyte Corporation), F001287 (Flexus Biosciences / Bristol-Myers Squibb), indoximod (NewLink Genetics), and NLG919 (NewLink Genetics). Epacadostat was described in PCT Publication WO 2010 / 005958, which is incorporated herein by reference in its entirety for all purposes. In one embodiment, the inhibitor is epacadostat. In another embodiment, the inhibitor is F001287. In another embodiment, the inhibitor is indoximod. In another embodiment, the inhibitor is NLG919.

[0237] In certain embodiments, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed herein is administered to an individual in combination with a vaccine. The vaccine may be, for example, a peptide vaccine, a DNA vaccine, or an RNA vaccine. In certain embodiments, the vaccine is a heat shock protein-based tumor vaccine or a heat shock protein-based pathogen vaccine. In certain embodiments, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed herein is administered to an individual in combination with a vaccine as described in WO 2016 / 183486 (e.g., a vaccine comprising at least one synthetic peptide comprising a cancer-specific mutation present in the individual's cancer), incorporated herein by reference in its entirety.In one specific embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed in this document is administered to an individual in combination with a heat shock protein-based tumor vaccine. Heat shock proteins (HSPs) are a family of highly conserved proteins found ubiquitously across all species. Their expression... Petition 870260065483, dated 03 / 07 / 2026, pages 136 / 214 126 / 195 can be strongly induced to much higher levels as a result of heat shock or other forms of stress, including exposure to toxins, oxidative stress, or glucose deprivation. Five families have been classified according to molecular weight: HSP-110, -90, -70, -60, and -28. HSPs distribute immunogenic peptides through the cross-presentation pathway onto antigen-presenting cells (APCs), such as macrophages and dendritic cells (DCs), leading to T cell activation. HSPs function as chaperone vehicles for tumor-associated antigenic peptides that form complexes capable of inducing tumor-specific immunity. After the release of dying tumor cells, HSP-antigen complexes are collected by antigen-presenting cells (APCs), where the antigens are processed into peptides that bind to MHC class I and class II molecules, leading to the activation of antitumor CD8+ and CD4+ T cells.The immunity produced by HSP complexes derived from tumor preparations is specifically targeted against the unique antigenic peptide repertoire expressed by each individual's cancer. Therefore, in one embodiment, the present invention relates to (a) an antibody and / or pharmaceutical composition of the present invention and (b) a vaccine for use as a medicament, for example, for use in a method for treating cancer. In one embodiment, the present invention relates to a pharmaceutical composition, kit or kit of parts comprising (a) an antibody and / or pharmaceutical composition of the present invention and (b) a vaccine. In one embodiment, the vaccine is a heat shock protein-based tumor vaccine. In one embodiment, the vaccine is a heat shock protein-based pathogen vaccine.

[0238] A heat shock protein-peptide complex (HSPPC) is a protein-peptide complex consisting of a heat shock protein non-covalently complexed with antigenic peptides. HSPPCs produce both inactive and adaptive immune responses. In a specific embodiment, Petition 870260065483, dated 03 / 07 / 2026, pages 137 / 214 127 / 195 The antigenic peptide (or peptides) exhibits antigenicity for the cancer being treated. HSPPCs are effectively sized by APCs via membrane receptors (primarily CD91) or by binding to Toll-like receptors. HSPPC internalization results in functional maturation of APCs with chemokine and cytokine production leading to activation of natural killer (NK) cells, monocytes, and Th1- and Th2-mediated immune responses. In certain embodiments, the HSPPCs used in methods disclosed herein comprise one or more heat shock proteins from the hsp60, hsp70, or hsp90 family of stress proteins complexed with antigenic peptides. In certain embodiments, the HSPPCs comprise hsc70, hsp70, hsp90, hsp110, grp170, gp96, calreticulin, or combinations of two or more thereof.

[0239] In a specific embodiment, the heat shock protein-peptide complex (HSPPC) comprises recombinant heat shock proteins (e.g., hsp70 or hsc70) or a peptide-binding domain thereof complexed with recombinant antigenic peptides. Recombinant heat shock proteins can be produced by recombinant DNA technology, for example, using human hsc70 sequence as described in Dwomiczak and Mirault, Nucleic Acids Res. 15:5181-5197 (1987) and GenBank accession number P11142 and / or Y00371, each of which is incorporated herein by reference in its entirety. In certain embodiments, the Hsp70 sequences are as described in Hunt and Morimoto Proc. Natl. Acad. Sci. EUA 82 (19), 64556459 (1985) and GenBank accession number P0DMV8 and / or M11717, each of which is incorporated herein by reference in its entirety.Antigenic peptides can also be prepared using recombinant DNA methods known in the art.

[0240] In certain embodiments, antigenic peptides comprise a Petition 870260065483, dated 03 / 07 / 2026, pp. 138 / 214 128 / 195 modified amino acid. In certain embodiments, the modified amino acid comprises a post-translational modification. In certain embodiments, the modified amino acid comprises a mimetic of a post-translational modification. In certain embodiments, the modified amino acid is a Tyr, Ser, Thr, Arg, Lys, or His that has been phosphorylated at a hydroxyl or amine side chain. In certain embodiments, the modified amino acid is a mimetic of a Tyr, Ser, Thr, Arg, Lys, or His amino acid that has been phosphorylated at a hydroxyl or amine side chain.

[0241] In a specific embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) disclosed herein is administered to an individual in combination with a heat shock protein-peptide complex (HSPPC), e.g., heat shock protein-peptide complex 96 (HSPPC-96), to treat cancer. HSPPC-96 comprises a 96 kDa heat shock protein (Hsp), gp96, complexed with antigenic peptides. HSPPC-96 is a cancer immunotherapy manufactured from an individual's tumor and contains the antigenic "fingerprint" of the cancer. In certain embodiments, this fingerprint contains unique antigens that are only present in those specific cancer cells of the particular individual, and the vaccine injection is intended to stimulate the individual's immune system to recognize and attack any cells with the specific cancer fingerprint.Therefore, in one embodiment, the present invention relates to an antibody and / or pharmaceutical composition of the present invention in combination with a heat shock protein and peptide complex (HSPPC) for use as a medicament and / or for use in a method for the treatment of cancer.

[0242] In certain modalities, HSPPC, for example, HSPPC-96, is produced from an individual's own tumor tissue. In one specific modality, HSPPC (e.g., HSPPC-96) is produced from a cancerous tumor or metastasis produced from the same that is being treated. In another specific modality, HSPPC (e.g., HSPPC-96) is autologous to the individual being treated. In certain Petition 870260065483, dated 03 / 07 / 2026, pp. 139 / 214 In 129 / 195 embodiments, the tumor tissue is non-necrotic tumor tissue. In certain embodiments, at least 1 gram (e.g., at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 grams) of non-necrotic tumor tissue is used to produce a vaccine regimen. In certain embodiments, after surgical resection, non-necrotic tumor tissue is frozen before use in vaccine preparation. In some embodiments, HSPPC, e.g., HSPPC-96, is isolated from tumor tissue by purification techniques, filtered, and prepared as an injectable vaccine. In certain embodiments, an individual receives 6 to 12 doses of HSPPC, e.g., HSPCC-96. In such modalities, doses of HSPPC, for example, HSPPC-96, can be administered weekly for the first 4 doses and then bi-weekly for the additional 2 to 8 doses.

[0243] Other examples of HSPPCs that can be used in accordance with the methods described in this document are disclosed in the following patents and patent applications, all of which are incorporated herein by reference in their entirety: U.S. Patents Nos. 6,391,306, 6,383,492, 6,403,095, 6,410,026, 6,436,404, 6,447,780, 6,447,781 and 6,610,659.

[0244] In certain embodiments, an anti-TIM-3 antibody disclosed herein is administered to an individual in combination with an adjuvant. Various adjuvants may be used depending on the treatment context. Non-limiting examples of suitable adjuvants include, but are not limited to, Complete Freund's Adjuvant (CFA), Incomplete Freund's Adjuvant (IFA), ISA montanide (incomplete Seppic adjuvant), the Ribi Adjuvant System (RAS), Titer Max, muramyl peptides, Syntex Adjuvant Formulation (SAF), alum (aluminum hydroxide and / or aluminum phosphate), aluminum salt adjuvants, Gerbu® adjuvants, nitrocellulose-absorbed antigen, encapsulated or captured antigen, monophosphoryl lipid Petition 870260065483, dated 03 / 07 / 2026, pages 140 / 214 130 / 195 3-De-O-acylated (3D-MPL), immunostimulatory oligonucleotides, toll-like receptor (TLR) ligands, mannan-binding lectin (MBL) ligands, STING agonists, immunostimulatory complexes such as saponins, Quil A, QS-21, QS7, ISCOMATRIX, and others. Other adjuvants include CpG oligonucleotides and double-stranded RNA molecules such as poly(A) and poly(U). Combinations of the above adjuvants may also be used. See, for example, U.S. Patents 6,645,495; 7,029,678; and 7,858,589, all of which are incorporated herein by reference in their entirety. In one embodiment, the adjuvant used in this document is QS-21 STIMULON.

[0245] In certain embodiments, an anti-TIM-3 antibody disclosed herein is administered to an individual in combination with an additional therapeutic agent comprising a TCR. In certain embodiments, the additional therapeutic agent is a soluble TCR. In certain embodiments, the additional therapeutic agent is a cell expressing a TCR. Therefore, in one embodiment, the present invention relates to an antibody and / or pharmaceutical composition of the present invention in combination with an additional therapeutic agent comprising a TCR for use as a medicament and / or for use in a method for the treatment of cancer.

[0246] In certain embodiments, an anti-TIM-3 antibody disclosed in this document is administered to an individual in combination with a cell expressing a chimeric antigen receptor (CAR). In certain embodiments, the cell is a T cell.

[0247] In certain embodiments, an anti-TIM-3 antibody disclosed in this document is administered to an individual in combination with a TCR mimetic antibody. In certain embodiments, the TCR mimetic antibody is an antibody that specifically binds to a peptide-MHC complex. For non-limiting examples of TCR mimetic antibodies, see, for example, U.S. Patent No. 9,074,000 and U.S. Publications Nos. 2009 / 0304679 A1 and 2014 / 0134191 A1, all of which are Petition 870260065483, dated 03 / 07 / 2026, pp. 141 / 214 131 / 195 incorporated into this document by reference in their entirety.

[0248] The anti-TIM-3 antibody (e.g., human TIM-3) and the additional therapeutic agent (e.g., chemotherapeutic, radiotherapeutic, checkpoint targeting agent, IDO inhibitor, vaccine, adjuvant, a soluble TCR, a cell expressing a TCR, a cell expressing a chimeric antigen receptor and / or a TCR mimetic antibody) may be administered separately, sequentially, or concomitantly as separate dosage forms. In one embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) is administered parenterally, and an IDO inhibitor is administered orally.

[0249] An antibody or pharmaceutical composition described herein may be delivered to an individual by a variety of routes. These include, but are not limited to, parenteral, intranasal, intratracheal, oral, intradermal, topical, intramuscular, intraperitoneal, transdermal, intravenous, intrathecal, intratumoral, conjunctival, intra-arterial, and subcutaneous routes. In certain embodiments, the antibody or pharmaceutical composition is delivered intravenously. Pulmonary administration may also be employed, for example, by the use of an inhaler or nebulizer and formulation with an aerosolizing agent for use as a spray. In certain embodiments, the antibody or pharmaceutical composition described herein is delivered subcutaneously or intravenously. In certain embodiments, the antibody or pharmaceutical composition described herein is delivered intra-arterially.In certain embodiments, the antibody or pharmaceutical composition described herein is delivered intratumorally. In certain embodiments, the antibody or composition described herein is delivered to a lymph node draining a tumor.

[0250] The amount of an antibody or compound that will be effective in treating and / or preventing a condition will depend on the nature of the disease and can be determined by standard clinical techniques. Petition 870260065483, dated 03 / 07 / 2026, pp. 142 / 214 132 / 195

[0251] The precise dose to be used in a compound will also depend on the route of administration and the severity of the infection or disease caused by the medication and should be decided according to the professional's judgment and the circumstances of each individual. For example, effective doses may also vary depending on the means of administration, target site, physiological state of the patient (including age, body weight and health), whether the patient is a human or an animal, other medications administered, or whether the treatment is prophylactic or therapeutic. Usually, the patient is a human, but non-human mammals, including transgenic mammals, may also be treated. Treatment dosages are ideally titrated to optimize safety and efficacy.

[0252] An anti-TIM-3 antibody (e.g., human TIM-3) described in this document can also be used to assess TIM-3 protein levels (e.g., human TIM-3) in a biological sample using classical immunohistological methods known to those skilled in the art, including immunoassays such as enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, or Western blotting. Suitable antibody assay markers are known in the art and include enzyme markers such as glucose oxidase; radioisotopes such as iodine (125I,121I), carbon (14C), sulfur (35S), tritium (3H), indium (121In), and technetium (99Tc); luminescent markers such as luminol; and fluorescent markers such as fluorescein, rhodamine, and biotin. Such markers can be used to identify an antibody described in this document.Alternatively, a second antibody that recognizes an anti-TIM-3 antibody (e.g., human TIM-3) described in this document can be identified and used in combination with an anti-TIM-3 antibody (e.g., human TIM-3) to detect TIM-3 protein levels (e.g., human TIM-3). Therefore, in one embodiment, the present invention relates to the use of an antibody of the present invention. Petition 870260065483, dated 03 / 07 / 2026, pp. 143 / 214 133 / 195 for in vitro detection of TIM-3 protein (e.g., human TIM-3) in a biological sample. In another embodiment, the present invention relates to the use of an anti-TIM-3 antibody of the invention to assess and / or detect levels of TIM-3 protein (e.g., human TIM-3) in a biological sample in vitro, optionally wherein the anti-TIM-3 antibody is conjugated to a radionuclide or detectable marker and / or carries a marker described in this document and / or wherein an immunohistological method is used.

[0253] The assessment of TIM-3 protein expression level (e.g., human TIM-3) is intended to include measuring or estimating quantitatively and qualitatively the TIM-3 protein level (e.g., human TIM-3) in a first biological sample directly (e.g., by determining or estimating the absolute protein level) or relatively (e.g., by comparing it to the disease-associated protein level in a second biological sample). The TIM-3 polypeptide expression level (e.g., human TIM-3) in the first biological sample can be measured or estimated and compared to a standard TIM-3 protein level (e.g., human TIM-3), where the standard is taken from a second biological sample obtained from an individual who does not have the disorder or is determined by averaging the levels of a population of individuals who do not have the disorder.As will be understood in the technique, once the “standard” TIM-3 polypeptide level (e.g., human TIM-3) is known, it can be repeatedly used as a standard for comparison. Therefore, in another embodiment, the present invention relates to an in vitro method for evaluating and / or detecting TIM-3 protein levels, for example, human TIM-3 protein levels, in a biological sample, comprising qualitatively or quantitatively measuring or estimating the TIM-3 protein level, for example, human TIM-3 protein, in a biological sample, by an immunohistological method.

[0254] As used in this document, the term “biological sample” Petition 870260065483, dated 03 / 07 / 2026, pages 144 / 214 134 / 195 refers to any biological sample obtained from an individual, cell line, tissue, or other cell source that potentially expresses TIM-3 (e.g., human TIM-3). Methods for obtaining tissue and body fluid biopsies from animals (e.g., humans) are well known in the art. Biological samples include peripheral blood mononuclear cells.

[0255] An anti-TIM-3 antibody (e.g., human TIM-3) described herein can be used for prognostic, diagnostic, monitoring, and screening applications, including in vitro and in vivo applications well known to those skilled in the art and based on this description. Prognostic, diagnostic, monitoring, and screening assays and kits for in vitro estimation and evaluation of immune system status and / or immune response can be used to predict, diagnose, and monitor to evaluate patient samples, including those known to have or suspected of having immune system dysfunction, or in relation to anticipated or desired immune system response, antigen response, or vaccine response.The estimation or assessment of immune system status and / or immune response is also useful for determining a patient's suitability for a clinical trial of a drug or for the administration of a particular chemotherapeutic agent, a radiotherapeutic agent, or an antibody, including combinations thereof, versus a different agent or antibody. This type of prognostic and diagnostic assessment and monitoring is already in practice using antibodies against the HER2 protein in breast cancer (HercepTest™, Dako), where the assay is also used to evaluate patients for antibody therapy with the use of Herceptin®. In vivo applications include targeted cell therapy and immune system modulation and radioimaging of immune responses. Therefore, in one embodiment, the present invention relates to an anti-TIM-3 antibody and / or pharmaceutical composition of the present invention for use as a diagnostic tool.In one embodiment, the present invention relates to an anti-TIM-3 antibody. Petition 870260065483, dated 03 / 07 / 2026, pp. 145 / 214 135 / 195 and / or pharmaceutical composition of the present invention for use in a method for predicting, diagnosing and / or monitoring an individual who has or is suspected of having an immune system dysfunction and / or in relation to an anticipated or desired immune system response, antigen response or vaccine response. In another embodiment, the present invention relates to the use of the anti-TIM-3 antibody of the invention, for predicting, diagnosing and / or monitoring an individual who has or is suspected of having an immune system dysfunction and / or in relation to an anticipated or desired immune system response, antigen response or vaccine response by evaluating and / or detecting TIM-3 protein levels in a biological sample from the individual in vitro.

[0256] In one embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) can be used in immunohistochemistry of biopsy samples. In one embodiment, the method is an in vitro method. In another embodiment, an anti-TIM-3 antibody (e.g., human TIM-3) can be used to detect levels of TIM-3 (e.g., human TIM-3) or levels of cells containing TIM-3 (e.g., human TIM-3) on their membrane surface, wherein the levels can then be linked to certain disease symptoms. The anti-TIM-3 antibodies (e.g., human TIM-3) described in this document may carry a detectable or functional marker and / or may be conjugated to a radionuclide or detectable marker.When fluorescence markers are used, currently available fluorescence-activated cell sorting and microscopy (FACS) analysis or a combination of both procedures from methods known in the art can be used to identify and quantify specific binding numbers. The anti-TIM-3 antibodies (e.g., human TIM-3) described herein may carry or be conjugated to a fluorescence marker. Exemplary fluorescence markers include, for example, reactive and conjugated probes, e.g., Amino. Petition 870260065483, dated 03 / 07 / 2026, pages 146 / 214 136 / 195 coumarin, fluorescein and Texas red, Alexa Fluor dyes, Cy dyes and DyLight dyes. An anti-TIM-3 antibody (e.g., human TIM-3) may carry or be conjugated to a radioactive tracer or radionuclide, such as the isotopes 3H, 14C, 32P, 35S, 36Cl, 51Cr, 57Co, 58Co, 59Fe, 67Cu, 90Y, 99Tc, 111In, 117Lu, 121I, 124I, 125I, 1311, 198Au, 211At, 213Bi, 225Ac and 186Re. When radioactive tracers are used, currently available counting procedures known in the art can be used to identify and quantify the specific binding of anti-TIM-3 antibody (e.g., human TIM-3) to TIM-3 (e.g., human TIM-3). In cases where the marker is an enzyme, detection can be performed using any of the colorimetric, spectrophotometric, fluorospectrophotometric, amperometric, or gasometric techniques currently known in the art.This can be achieved by placing a sample or a control sample in contact with an anti-TIM-3 antibody (e.g., human TIM-3) under conditions that allow the formation of a complex between the antibody and TIM-3 (e.g., human TIM-3). Any complexes formed between the antibody and TIM-3 (e.g., human TIM-3) are detected and compared in the sample and the control. In light of the specific binding of the antibodies described herein to TIM-3 (e.g., human TIM-3), the antibodies can be used to specifically detect the expression of TIM-3 (e.g., human TIM-3) on the cell surface. The antibodies described herein can also be used to purify TIM-3 (e.g., human TIM-3) by means of immunoaffinity purification.Also included in this document is an assay system that can be prepared in the form of a test kit, kit or parts kit for the quantitative analysis of the extent of the presence of, for example, TIM-3 (e.g., human TIM-3) or TIM-3 (e.g., human TIM-3) / TIM-3 ligand (e.g., human TIM-3) complexes. The system, test kit, kit or parts kit may comprise an identified component, e.g., an identified antibody, and one or more additional immunochemical reagents. Petition 870260065483, dated 03 / 07 / 2026, pages 147 / 214 137 / 195 6.5 Polynucleotides, Vectors and Methods for Producing Anti-TIM-3 Antibodies

[0257] In another aspect, polynucleotides comprising a nucleotide sequence encoding an antibody described herein or a fragment thereof (e.g., a variable light chain region and / or a variable heavy chain region) that specifically binds to a TIM-3 antigen (e.g., human TIM-3) and vectors, e.g., vectors comprising such polynucleotides for recombinant expression in host cells (e.g., E. coli and mammalian cells), are provided herein. Polynucleotides comprising nucleotide sequences encoding a heavy and / or light chain of any of the antibodies provided herein are provided herein, as well as vectors comprising such polynucleotide sequences, e.g., expression vectors for their effective expression in host cells, e.g., mammalian cells.

[0258] As used in this document, an “isolated” polynucleotide or nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the natural source (e.g., in a mouse or a human) of the nucleic acid molecule. In addition, an “isolated” nucleic acid molecule, such as a cDNA molecule, may be substantially free of other cellular material or culture medium when produced by recombinant techniques or substantially free of chemical precursors or other chemicals when chemically synthesized. For example, the term “substantially free” includes preparations of polynucleotide or nucleic acid molecules that have less than about 15%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (in particular less than about 10%) of other material, for example, cellular material, culture medium, other nucleic acid molecules, chemical precursors, and / or other chemicals.In one specific embodiment, a nucleic acid molecule (or molecules) that... Petition 870260065483, dated 03 / 07 / 2026, pp. 148 / 214 138 / 195 encodes an antibody described in this document and is isolated or purified.

[0259] In particular aspects, polynucleotides comprising antibody-coding nucleotide sequences are provided herein, which bind specifically to a TIM-3 polypeptide (e.g., human TIM-3) and comprise an amino acid sequence as described herein, as well as antibodies that compete with such antibodies for binding to a TIM-3 polypeptide (e.g., human TIM-3) (e.g., in a dose-dependent manner) or that bind to the same epitope as that of such antibodies.

[0260] In certain respects, polynucleotides comprising a nucleotide sequence encoding either the light chain or the heavy chain of an antibody described herein are provided in this document. The polynucleotides may comprise nucleotide sequences encoding a light chain comprising VL FRs and CDRs of antibodies described herein (see, for example, Table 1) or nucleotide sequences encoding a heavy chain comprising VH FRs and CDRs of antibodies described herein (see, for example, Table 1).

[0261] Also provided in this document are polynucleotides encoding an anti-TIM-3 antibody (e.g., human TIM-3) that are optimized, for example, by codon / RNA optimization, substitution with heterologous signaling sequences, and elimination of mRNA instability elements. Methods for generating optimized nucleic acids encoding an anti-TIM-3 antibody (e.g., human TIM-3) or a fragment thereof (e.g., light chain, heavy chain, VH domain, or VL domain) for recombinant expression by introducing codon changes and / or eliminating inhibitory regions in the mRNA can be performed by adapting the optimization methods described, for example, Petition 870260065483, dated 03 / 07 / 2026, pages 149 / 214 139 / 195 in U.S. Patents Nos. 5,965,726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498, consequently, all of which are incorporated herein by reference in their entirety. For example, potential splicing sites and instability elements (e.g., A / T-rich or A / U-rich elements) within RNA can undergo mutation without altering the amino acids encoded by the nucleic acid sequences to increase RNA stability for recombinant expression. The alterations use genetic code degeneracy, for example, with the use of an alternative codon for an identical amino acid. In some embodiments, it may be desirable to alter one or more codons to encode a conservative mutation, for example, a similar amino acid with chemical structure and properties and / or function as the original amino acid.Such methods can increase the expression of an anti-TIM-3 antibody (e.g., human TIM-3) or fragment thereof by at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold or more relative to the expression of an anti-TIM-3 antibody (e.g., human TIM-3) encoded by non-optimized polynucleotides.

[0262] In certain embodiments, an optimized polynucleotide sequence encoding an anti-TIM-3 antibody (e.g., human TIM-3) described herein or a fragment thereof (e.g., VL domain and / or VH domain) can hybridize to an antisense polynucleotide (e.g., complementary) of a non-optimized polynucleotide sequence encoding an anti-TIM-3 antibody (e.g., human TIM-3) described herein or a fragment thereof (e.g., VL domain and / or VH domain). In specific embodiments, an optimized nucleotide sequence encoding an anti-TIM-3 antibody (e.g., human TIM-3) described herein or a fragment hybridizes under high stringency conditions to an antisense polynucleotide of a non-optimized polynucleotide sequence encoding an antibody Petition 870260065483, dated 03 / 07 / 2026, pages 150 / 214 140 / 195 anti-TIM-3 (e.g., human TIM-3) described herein or a fragment thereof. In a specific embodiment, an optimized nucleotide sequence encoding an anti-TIM-3 antibody (e.g., human TIM-3) described herein or a fragment thereof hybridizes under high-stringency, intermediate, or lower-stringency hybridization conditions to an antisense polynucleotide of a non-optimized nucleotide sequence encoding an anti-TIM-3 antibody (e.g., human TIM-3) described herein or a fragment thereof. Information on hybridization conditions has been described; see, for example, U.S. Patent Application Publication No. 2005 / 0048549 (e.g., paragraphs 72 to 73), which is incorporated herein by reference in its entirety.

[0263] Polynucleotides can be obtained and the nucleotide sequence of the polynucleotides determined by any method known in the art. The nucleotide sequences encoding the antibodies described in this document, for example, antibodies described in Table 1, and modified versions of these antibodies can be determined using methods well known in the art, for example, nucleotide codons known to encode particular amino acids are assembled in such a way as to generate a nucleic acid encoding the antibody. Such an antibody-encoding polynucleotide can be assembled from chemically synthesized oligonucleotides (for example, as described in Kutmeier G et al., (1994), BioTechniques 17: 242-246, incorporated herein by reference in its entirety), which, in summary, involves the synthesis of overlapping oligonucleotides containing portions of the antibody-coding sequence, annealing and ligation of these oligonucleotides, and then amplification of the linked oligonucleotides by PCR.

[0264] Alternatively, a polynucleotide encoding an antibody described Petition 870260065483, dated 03 / 07 / 2026, pages 151 / 214 141 / 195 in this document can be generated from nucleic acid from a suitable source (e.g., a hybridoma) using methods well-known in the art (e.g., PCR and other molecular cloning methods). For example, PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of a known sequence can be performed using genomic DNA obtained from hybridoma cells that produce the antibody of interest. Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the light chain and / or heavy chain of an antibody. Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the variable light chain region and / or variable heavy chain region of an antibody.The amplified nucleic acids can be cloned into vectors for expression in host cells and for further cloning, for example, to generate chimeric or humanized antibodies.

[0265] If a clone containing a nucleic acid encoding a particular antibody is not available, but the sequence of the antibody molecule is known, a nucleic acid encoding the antibody may be chemically synthesized or obtained from a suitable source (e.g., an antibody cDNA library or a cDNA library generated from, or nucleic acid, preferably poly A+RNA, isolated from any tissue or cells expressing the antibody or such as hybridoma cells selected to express an antibody described in this document) by PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of the sequence or cloning using an oligonucleotide probe specific for the particular gene sequence to identify, for example, a cDNA clone from a cDNA library encoding the antibody.The amplified nucleic acids generated by PCR can then be cloned into replicable cloning vectors using any well-known method in the art. Petition 870260065483, dated 03 / 07 / 2026, pages 152 / 214 142 / 195

[0266] The DNA encoding anti-TIM-3 antibodies (e.g., human TIM-3) described in this document can be readily isolated and sequenced using conventional procedures (e.g., using oligonucleotide probes that are capable of specifically binding to genes encoding the heavy and light chains of anti-TIM-3 antibodies (e.g., human TIM-3)). Hybridoma cells can serve as a source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells, such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells (e.g., CHO GS System™ (Lonza) CHO cells), or myeloma cells that do not otherwise produce immunoglobulin protein, to achieve the synthesis of recombinant anti-TIM-3 antibodies (e.g., human TIM-3) in host cells.

[0267] To generate whole antibodies, PCR primers including VH or VL nucleotide sequences, a restriction site, and a flanking sequence to protect the restriction site can be used to amplify the VH or VL sequences into scFv clones. Using sets of cloning procedures known to those skilled in the art, the PCR-amplified heavy chain domains can be cloned into vectors expressing a VH constant region, for example, the human gamma 4 constant region, and the PCR-amplified VL domains can be cloned into vectors expressing a light chain constant region, for example, the human kappa or lambda constant regions. In certain embodiments, the vectors to express the VH or VL domains comprise an EF-M promoter, a secretion signal, a cloning site for the variable region, constant domains, and a selection marker such as neomycin.The VH and VL domains can also be cloned into a vector that expresses the necessary constant regions. The heavy chain conversion vectors and light chain conversion vectors are then co-transfected into cell lines to generate cell lines. Petition 870260065483, dated 03 / 07 / 2026, pages 153 / 214 143 / 195 stable or transient cells expressing full-length antibodies, for example, IgG, using sets of procedures known to those skilled in the art.

[0268] DNA can also be modified, for example, by using the coding sequence for human light and heavy chain constant domains instead of murine sequences, or by covalently joining all or part of the coding sequence for a non-immunoglobulin polypeptide to the immunoglobulin coding sequence.

[0269] Polynucleotides are also provided that hybridize under high-stringency, intermediate-stringency, or less-stringency hybridization conditions to polynucleotides encoding an antibody described herein. In specific embodiments, the polynucleotides described herein hybridize under high-stringency, intermediate-stringency, or less-stringency hybridization conditions to polynucleotides encoding a VH domain and / or VL domain provided herein.

[0270] The hybridization conditions have been described in the art and are known to one skilled in the art. For example, hybridization under stringent conditions may involve hybridization to filter DNA bound in 6x sodium chloride / sodium citrate (SSC) at about 45°C followed by one or more washes in 0.2x SSC / 0.1% SDS at about 50 to 65°C; hybridization under highly stringent conditions may involve hybridization to filter nucleic acid bound in 6x SSC at about 45°C followed by one or more washes in 0.1x SSC / 0.2% SDS at about 68°C. Hybridization under other stringent hybridization conditions is known to those skilled in the art and has been described, see, for example, Ausubel FM et al., eds., (1989) Current Protocols in Molecular Biology, Vol. I, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York on pages 6.3.1-6.3.6 and 2.10.3, which is incorporated herein by reference in its entirety. Petition 870260065483, dated 03 / 07 / 2026, pp. 154 / 214 144 / 195

[0271] In certain aspects, cells (e.g., host cells) expressing (e.g., recombinantly) antibodies described herein that bind specifically to TIM-3 (e.g., human TIM-3) and related expression polynucleotides and vectors are provided herein. Vectors (e.g., expression vectors) comprising polynucleotides comprising nucleotide sequences encoding anti-TIM-3 antibodies (e.g., human TIM-3) or a fragment for recombinant expression in host cells, preferably mammalian cells, are provided herein. Host cells comprising such vectors for recombinantly expressing anti-TIM-3 antibodies (e.g., human TIM-3) described herein (e.g., humanized or human antibody) are also provided herein.In one particular aspect, methods are provided in this document for producing an antibody described herein, comprising expressing such antibody from a host cell.

[0272] Recombinant expression of an antibody described herein (e.g., a full-length antibody, heavy and / or light chain antibody, or a single-chain antibody described herein) that specifically binds to TIM-3 (e.g., human TIM-3) involves constructing an expression vector containing a polynucleotide encoding the antibody. Once a polynucleotide encoding an antibody molecule, heavy and / or light chain antibody, or a fragment thereof (e.g., variable regions of heavy and / or light chains) described herein has been obtained, the vector for producing the antibody molecule can be produced using recombinant DNA technology with procedures known in the art. In this way, methods for preparing a protein after expression of a polynucleotide containing an antibody or antibody fragment (e.g., chain Petition 870260065483, dated 03 / 07 / 2026, pp. 155 / 214 145 / 195 light or heavy chain) encoding nucleotide sequences are described in this document. Methods well known to those skilled in the art can be used to construct expression vectors containing antibody or antibody fragment (e.g., light chain or heavy chain) encoding appropriate translational and transcriptional control sequences and signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Replicable vectors are also provided comprising a nucleotide sequence encoding an antibody molecule described in this document, a heavy or light chain of an antibody, a variable region of a heavy or light chain of an antibody or a fragment thereof, or a CDR of a heavy or light chain, operationally linked to a promoter.Such vectors may include, for example, the nucleotide sequence encoding the constant region of the antibody molecule (see, for example, international publications WO 86 / 05807 and WO 89 / 01036; and US patent 5,122,464, which are incorporated herein by reference in their entirety) and variable regions of the antibody may be cloned into such a vector for expression of the entire light chain, the entire heavy chain, or both the light chain and the entire heavy chain.

[0273] An expression vector can be transferred into a cell (e.g., host cell) by conventional techniques, and the resulting cells can then be cultured by conventional techniques to produce an antibody described herein or a fragment thereof. In this way, host cells containing a polynucleotide encoding an antibody described herein or fragments thereof, or a heavy or light chain thereof, or a fragment thereof, or a single-chain antibody described herein, operationally linked to a Petition 870260065483, dated 03 / 07 / 2026, pages 156 / 214 146 / 195 promoter for the expression of such sequences in the host cell. In certain embodiments for the expression of double-chain antibodies, vectors encoding both the heavy and light chains individually can be co-expressed in the host cell for expression of the entire immunoglobulin molecule, as detailed below. In certain embodiments, a host cell contains a vector comprising a polynucleotide encoding either the heavy chain or the light chain of an antibody described herein, or a fragment thereof.In specific embodiments, a host cell contains two different vectors: a first vector comprising a polynucleotide encoding a heavy chain or a variable region of a heavy chain of an antibody described herein, or a fragment thereof; and a second vector comprising a polynucleotide encoding a light chain or a variable region of a light chain of an antibody described herein, or a fragment thereof. In other embodiments, a first host cell comprises a first vector comprising a polynucleotide encoding a heavy chain or a variable region of a heavy chain of an antibody described herein, or a fragment thereof, and a second host cell comprises a second vector comprising a polynucleotide encoding a light chain or a variable region of a light chain of an antibody described herein.In specific embodiments, a heavy chain / variable heavy chain region expressed by a first cell has associated with a light chain / variable light chain region of a second cell to form an anti-TIM-3 antibody (e.g., human TIM-3) described herein. In certain embodiments, a host cell population comprising such a first host cell and such a second host cell is provided herein.

[0274] In a particular form, a is provided in this document Petition 870260065483, dated 03 / 07 / 2026, pages 157 / 214 147 / 195 vector population comprising a first vector comprising a polynucleotide encoding a light chain / variable light chain region of an anti-TIM-3 antibody (e.g., human TIM-3) described herein, and a second vector comprising a polynucleotide encoding a heavy chain / variable heavy chain region of an anti-TIM-3 antibody (e.g., human TIM-3) described herein.

[0275] A variety of host expression vector systems can be used to express antibody molecules described herein (see, for example, U.S. Patent No. 5,807,715, which is incorporated herein by reference in its entirety). Such host expression systems represent vehicles by which coding sequences of interest can be produced and subsequently purified, but they also represent cells that, when transformed or transfected with the appropriate nucleotide coding sequences, can express an antibody molecule described herein in situ. These include, but are not limited to, microorganisms such as bacteria (e.g., E. coli and B.subtilis) transformed with recombinant plasmid DNA, cosmid DNA, or bacteriophage DNA expression vectors containing antibody-coding sequences; yeast (e.g., Saccharomyces, Pichia) transformed with recombinant yeast expression vectors containing antibody-coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing antibody-coding sequences; plant cell systems (e.g., green algae such as Chlamydomonas reinhardtii) infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing antibody-coding sequences; or mammalian cell systems. Petition 870260065483, dated 03 / 07 / 2026, pages 158 / 214 148 / 195 (e.g., COS cells (e.g., COS1 or COS), CHO, BHK, MDCK, HEK 293, NS0, PER.C6, VERO, CRL7O3O, HsS78Bst, HeLa, and NIH 3T3, HEK293T, HepG2, SP210, Rl.l, BW, LM, BSC1, BSC40, YB / 20, and BMT10 cells) harboring recombinant expression constructs containing promoters derived from mammalian cell genomes (e.g., metallothionein promoter) or mammalian viruses (e.g., adenovirus late promoter; vaccinia virus 7.5K promoter). In specific embodiments, the antibody-expressing cells described herein are CHO cells, e.g., CHO GS System™ CHO cells (Lonza). In one particular embodiment, the antibody-expressing cells described herein are human cells, for example, human cell lines. In another specific embodiment, a mammalian expression vector is pOptiVEC™ or pcDNA3.3.In one particular embodiment, bacterial cells such as Escherichia coli, or eukaryotic cells (e.g., mammalian cells), especially for the expression of whole recombinant antibody molecules, are used for the expression of a recombinant antibody molecule. For example, mammalian cells such as Chinese hamster ovary (CHO) cells, in conjunction with a vector such as the early intermediate major gene promoter element of human cytomegalovirus, are an effective expression system for antibodies (Foecking MK & Hofstetter H (1986) Gene 45: 101-105; and Cockett MI et al., (1990) Biotechnology 8(7): 662-667; each of which is incorporated herein by reference in its entirety). In certain embodiments, the antibodies described herein are produced by CHO cells or NSO cells.In one specific embodiment, the expression of nucleotide sequences encoding antibodies described herein that bind specifically to TIM-3 (e.g., human TIM-3) is regulated by a constitutive promoter, an inducible promoter, or a tissue-specific promoter.

[0276] In bacterial systems, numerous expression vectors can be Petition 870260065483, dated 03 / 07 / 2026, pp. 159 / 214 149 / 195 advantageously selected depending on the intended use for the antibody molecule being expressed. For example, when a large quantity of such an antibody must be produced, for the generation of pharmaceutical compositions of an antibody molecule, vectors that direct the expression of high levels of fusion protein products that are readily purified may be desirable. Such vectors include, but are not limited to, the E. coli pUR278 expression vector (Ruether U & Mueller-Hill B (1983) EMBO J 2: 1791-1794), in which the antibody coding sequence can be individually ligated to the vector in the frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye S & Inouye M (1985) Nuc Acids Res 13: 3101-3109; Van Heeke G & Schuster SM (1989) J Biol Chem 24: 5503-5509); and similar documents, all of which are incorporated into this document by reference in their entirety.For example, pGEX vectors can also be used to express foreign polypeptides such as fusion proteins with glutathione 5-transferase (GST). In general, such fusion proteins are soluble and can be easily purified from lysed cells by adsorption and binding to matrix glutathione-agarose microspheres followed by elution in the presence of free glutathione. pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned gene product can be released from the GST portion.

[0277] In an insect system, the Autographa californica nuclear polyhedrosis virus (AcNPV), for example, can be used as a vector to express foreign genes. The virus proliferates in Spodoptera frugiperda cells. The antibody-coding sequence can be individually cloned into non-essential regions (e.g., the polyhedrin gene) of the virus and placed under the control of an AcNPV promoter (e.g., the polyhedrin promoter).

[0278] In mammalian host cells, numerous virus-based expression systems can be used. In cases where an adenovirus is used Petition 870260065483, dated 03 / 07 / 2026, pages 160 / 214 Using a 150 / 195 expression vector, the antibody-coding sequence of interest can be ligated to an adenovirus transcription / translation control complex, for example, the late promoter and the tripartite leader sequence. This chimeric gene can then be inserted into the adenovirus genome by in vitro or in vivo recombination. Insertion into a non-essential region of the viral genome (e.g., E1 or E3 region) will result in a recombinant virus that is viable and capable of expressing the antibody molecule in infected hosts (e.g., see Logan J & Shenk T (1984) PNAS 81(12): 3655-3659, which is incorporated herein by reference in its entirety). Specific initiation signals may also be required for efficient translation of inserted antibody-coding sequences. These signals include the ATG initiation codon and adjacent sequences.Furthermore, the initiation codon needs to be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression can be enhanced by the inclusion of appropriate transcription enhancers, transcription terminators, etc. (see, for example, Bitter G et al., (1987) Methods Enzymol. 153: 516-544, which is incorporated herein by reference in its entirety).

[0279] Furthermore, a host cell strain can be chosen that modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific desired form. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products can be important for protein function. Different host cells have specific characteristics and mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure correct protein modification and processing. Petition 870260065483, dated 03 / 07 / 2026, pages 161 / 214 151 / 195 strange expressed. For this purpose, eukaryotic host cells that possess the cellular mechanisms for the appropriate processing of the primary transcript, glycosylation and phosphorylation of the gene product can be used. These mammalian host cells include, but are not limited to, CHO, VERO, BHK, Hela, MDCK, HEK 293, NIH 3T3, W138, BT483, Hs578T, HTB2, BT2O and T47D cells, NS0 (a murine myeloma cell line that does not endogenously produce any immunoglobulin chain), CRL7O3O, COS (e.g., COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, R1.1, BW, L-M, BSC1, BSC40, YB / 20, BMT10 and HsS78Bst cells. In certain embodiments, the anti-TIM-3 antibodies (e.g., human TIM-3) described herein are produced in mammalian cells, such as CHO cells.

[0280] In a specific embodiment, the antibodies described herein have reduced fucose content or no fucose content. Such antibodies can be produced using techniques known in the art. For example, the antibodies can be expressed in cells deficient or lacking the ability to fucosylate. In a specific example, cell lines with a knockout of both α1,6-fucosyltransferase alleles can be used to produce antibodies with reduced fucose content. The Potelligent® (Lonza) system is an example of such a system that can be used to produce antibodies with reduced fucose content.

[0281] For long-term high-throughput production of recombinant proteins, stable expression cells can be generated. For example, cell lines that stably express an anti-TIM3 antibody (e.g., human TIM-3) described in this document can be manipulated. In specific embodiments, a cell provided in this document stably expresses a light chain / variable light chain region and a heavy chain / variable heavy chain region that associate to form an antibody described in this document. Petition 870260065483, dated 03 / 07 / 2026, pages 162 / 214 152 / 195

[0282] In certain respects, instead of using expression vectors containing viral origins of replication, host cells can be transformed with DNA controlled through appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.) and a selectable marker. After the introduction of the foreign DNA / polynucleotide, the manipulated cells can be allowed to proliferate for 1–2 days in an enriched medium and then transferred to a selective medium. The selectable marker on the recombinant plasmid confers resistance to selection and allows the cells to stably integrate into the plasmid within their chromosomes and proliferate to form foci that can in turn be cloned and expanded into cell lines.This method can be advantageously used to manipulate cell lines that express an anti-TIM-3 antibody (e.g., human TIM-3) described in this document or a fragment thereof. Such cell lines can be particularly useful in screening and evaluating compositions that interact directly or indirectly with the antibody molecule.

[0283] Several selection systems can be used which include, but are not limited to, the herpes simplex virus genes herpes thymidine kinase (Wigler M et al., (1977) Cell 11(1): 223-232), hypoxanthine-guanine phosphoribosyltransferase (Szybalska EH & Szybalski W (1962) PNAS 48(12): 2026-2034) and adenine phosphoribosyltransferase (Lowy I et al., (1980) Cell 22(3): 817-823) in tk, hgprt or aprt cells, respectively, all of which are incorporated herein by reference in their entirety. Furthermore, antimetabolite resistance can be used as the basis for selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler M et al., (1980) PNAS 77(6): 3567-3570; O'Hare K et al., (1981) PNAS 78: 1527-1531); gpt, which confers resistance to mycophenolic acid (Mulligan RC & Berg P (1981) PNAS 78(4): 2072-2076); neo, which confers resistance to aminoglycoside G-418 (Wu GY & Wu CH (1991) Biotherapy 3: 87-95; Tolstoshev P (1993) Ann Petition 870260065483, dated 03 / 07 / 2026, pp. 163 / 214 153 / 195 Rev Pharmacol Toxicol 32: 573-596; Mulligan RC (1993) Science 260: 926-932; and Morgan RA & Anderson WF (1993) Ann Rev Biochem 62: 191-217; Nabel GJ & Feigner PL (1993) Trends Biotechnol 11(5): 211-215); and hygro, which confers resistance to hygromycin (Santerre RF et al., (1984) Gene 30(1-3): 147-156), each of which is incorporated herein by reference in its entirety. The methods commonly known in recombinant DNA technology can be routinely applied to select the desired recombinant clone and such methods are described, for example, in Ausubel FM et al., (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); Kriegler M, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13, Dracopoli NC et al., (eds.), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994); Colbere-Garapin F et al., (1981) J Mol Biol 150: 1-14, each of which is incorporated herein by reference in its entirety.

[0284] The expression levels of an antibody molecule can be increased by vector amplification (for an analysis, see Bebbington CR & Hentschel CCG, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3 (Academic Press, New York, 1987), which is incorporated herein by reference in its entirety). When a marker in the antibody-expressing vector system is amplifiable, increasing the level of inhibitor present in the host cell culture will increase the number of copies of the marker gene. Since the amplified region is associated with the antibody gene, antibody production will also increase (Crouse GF et al., (1983) Mol Cell Biol 3: 257-266, which is incorporated herein by reference in its entirety.

[0285] The host cell can be cotransfected with two or more expression vectors described in this document, where the first vector encodes a Petition 870260065483, dated 03 / 07 / 2026, pages 164 / 214 The first vector encodes a heavy chain-derived polypeptide, while the second vector encodes a light chain-derived polypeptide. The two vectors may contain identical selectable markers that enable equal expression of heavy and light chain polypeptides. Host cells may be co-transfected with different amounts of the two or more expression vectors. For example, host cells may be transfected with any of the following ratios of a first expression vector and a second expression vector: 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, or 1:50.

[0286] Alternatively, a single vector that encodes and is capable of expressing both heavy and light chain polypeptides may be used. In such situations, the light chain should be placed before the heavy chain to avoid an excess of toxic free heavy chain (Proudfoot NJ (1986) Nature 322: 562-565; and Kohler G (1980) PNAS 77: 2197-2199; each of which is incorporated herein by reference in its entirety). The coding sequences for the heavy and light chains may comprise cDNA or genomic DNA. The expression vector may be monocistronic or multicistronic. A multicistronic nucleic acid construct can encode 2, 3, 4, 5, 6, 7, 8, 9, 10 or more genes / nucleotide sequences, or in the range of 2 to 5, 5 to 10, or 10 to 20 genes / nucleotide sequences.For example, a bicistronic nucleic acid construct might comprise, in the following order, a promoter, a first gene (e.g., the heavy chain of an antibody described herein), and a second gene (e.g., the light chain of an antibody described herein). In such an expression vector, transcription of both genes can be driven by the promoter, while translation of mRNA from the first gene can occur via a termination-dependent scan mechanism, and translation of mRNA from the second gene can occur via a termination-independent mechanism, for example, by an IRES. Petition 870260065483, dated 03 / 07 / 2026, pages 165 / 214 155 / 195

[0287] Once an antibody molecule described herein has been produced by recombinant expression, it can be purified by any method known in the art for the purification of an immunoglobulin molecule, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after protein A, and size column chromatography), centrifugation, differential solubility, or by any other standard technique for protein purification. Additionally, the antibodies described herein can be fused to heterologous polypeptide sequences described herein or otherwise known in the art to facilitate purification.

[0288] In specific embodiments, an antibody described herein is isolated or purified. In general, an isolated antibody is one that is substantially free of other antibodies with antigenic specificities different from the isolated antibody. For example, in a particular embodiment, a preparation of an antibody described herein is substantially free of cellular material and / or chemical precursors. The term “substantially free of cellular material” includes preparations of an antibody in which the antibody is separated from the cellular components of the cells from which it is isolated or recombinantly produced.Thus, an antibody that is substantially free of cellular material includes antibody preparations that have less than about 30%, 20%, 10%, 5%, 2%, 1%, 0.5%, or 0.1% (by dry weight) of heterologous protein (also referred to herein as a “contaminant protein”) and / or variants of an antibody, for example, different post-translational modified forms of an antibody or other different versions of an antibody (e.g., antibody fragments). When the antibody is produced recombinantly, it is also generally substantially free of culture medium, that is, the culture medium represents less than about 20%, 10%, 2%, 1%, 0.5%, or 0.1% of the volume of the preparation. Petition 870260065483, dated 03 / 07 / 2026, pp. 166 / 214 156 / 195 Protein preparation. When the antibody is produced by chemical synthesis, it is generally substantially free of chemical precursors or other chemicals, that is, it is separated from the chemical precursors or other chemicals involved in protein synthesis. Correspondingly, such preparations of an antibody have less than about 30%, 20%, 10%, or 5% (by dry weight) of chemical precursors or compounds other than the antibody of interest. In one particular embodiment, the antibodies described herein are isolated or purified.

[0289] Antibodies or fragments thereof that bind specifically to TIM-3 (e.g., human TIM-3) can be produced by any method known in the art for antibody synthesis, for example, by chemical synthesis or by recombinant expression techniques. The methods described herein employ, unless otherwise indicated, conventional sets of procedures in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields within the skill of the art. These sets of procedures are described, for example, in the references cited herein and are fully explained in the literature. See, for example, Maniatis T et al., (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Sambrook J et al., (1989), Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press; Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel FM et al., Current Protocols in Molecular Biology, John Wiley & Sons (1987 and annual updates); Current Protocols in Immunology, John Wiley & Sons (1987 and annual updates) Gait (ed.) (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; Eckstein (ed.) (1991) Oligonucleotide and Analogs: A Practical Approach, IRL Press; Birren B et al., (eds.). Petition 870260065483, dated 03 / 07 / 2026, pp. 167 / 214 157 / 195 (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press, all of which are incorporated herein by reference in their entirety.

[0290] In a specific embodiment, an antibody described in this document is an antibody (e.g., recombinant antibody) prepared, expressed, created, or isolated by any means involving creation, for example, through synthesis, genetic engineering of DNA sequences. In certain embodiments, such an antibody comprises sequences (e.g., DNA sequences or amino acid sequences) that do not naturally exist within the germline antibody repertoire of an animal or mammal (e.g., human) in vivo.

[0291] In one aspect, a method is provided herein for manufacturing an antibody that specifically binds to TIM-3 (e.g., human TIM-3) comprising culturing a cell or host cell described herein. In one embodiment, the method is performed in vitro. In a certain aspect, a method is provided herein for manufacturing an antibody that specifically binds to TIM-3 (e.g., human TIM-3) comprising expressing (e.g., recombinantly expressing) the antibody using a cell or host cell described herein (e.g., a cell or host cell comprising polynucleotides encoding an antibody described herein). In a particular embodiment, the cell is a single cell. In a particular embodiment, exogenous polynucleotides have been introduced into the cell.In one particular embodiment, the method additionally includes the step of purifying the antibody obtained from the host cell or cell.

[0292] Methods for producing polyclonal antibodies are known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th edition, Ausubel FM et al., eds., John Wiley and Sons, New York, which is incorporated herein by reference in its entirety). Petition 870260065483, dated 03 / 07 / 2026, pp. 168 / 214 158 / 195

[0293] Monoclonal antibodies can be prepared using a wide variety of known sets of procedures in the art, including the use of phage presentation, recombinant, and hybridoma sets of procedures, or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma sets of procedures, including those known in the art and taught, for example, in E & Lane D, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nd edition (1988); Hammerling GJ et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY, 1981), each of which is incorporated herein by reference in its entirety. The term “monoclonal antibody,” as used herein, is not limited to antibodies produced through hybridoma technology.For example, monoclonal antibodies can be produced recombinantly from host cells that exogenously express an antibody described in this document or a fragment thereof, for example, the light chain and / or heavy chain of such antibody.

[0294] In specific embodiments, a “monoclonal antibody,” as used herein, is an antibody produced by a single cell (e.g., hybridoma or host cell producing a recombinant antibody), wherein the antibody binds specifically to TIM-3 (e.g., human TIM-3) as determined, for example, by ELISA or other competitive binding or antigen-binding assay known in the art or in examples provided herein. In particular embodiments, a monoclonal antibody may be a chimeric antibody or a humanized antibody. In certain embodiments, a monoclonal antibody is a monovalent antibody or a multivalent (e.g., bivalent) antibody. In particular embodiments, a monoclonal antibody is a monospecific or multispecific (e.g., bispecific) antibody. The monoclonal antibodies described herein may be produced, for example, by Petition 870260065483, dated 03 / 07 / 2026, pp. 169 / 214 159 / 195 hybridoma method as described in Kohler G & Milstein C (1975) Nature 256: 495, which is incorporated herein by reference in its entirety, or may, for example, be isolated from phage libraries using the techniques described herein. Other methods for the preparation of clonal cell lines and monoclonal antibodies expressed through them are well known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th edition, Ausubel FM et al., supra).

[0295] The methods for producing and screening for specific antibodies using hybridoma technology are routine and well-known in the art. For example, in the hybridoma method, a mouse or other suitable host animal, such as a sheep, goat, rabbit, rat, hamster, or monkey, is immunized to elicit lymphocytes that produce or have the capacity to produce antibodies that will specifically bind to the protein (e.g., TIM-3 (e.g., human TIM-3)) used for immunization. Alternatively, lymphocytes can be immunized in vitro. The lymphocytes are then fused with myeloma cells using a suitable fusion agent, such as polyethylene glycol, to form a hybridoma cell (Goding JW (Ed), Monoclonal Antibodies: Principles and Practice, pages 59-103 (Academic Press, 1986), incorporated herein by reference in its entirety).Additionally, a set of RIMMS (repetitive multiple immunization sites) procedures can be used to immunize an animal (Kilpatrick KE et al., (1997) Hybridoma 16:381-389, incorporated herein by reference in its entirety).

[0296] In some embodiments, mice (or other animals such as rats, monkeys, gorillas, pigs, sheep, hamsters or dogs) can be immunized with an antigen (e.g., TIM-3 (e.g., human TIM-3)) and once an immune response is detected, for example, specific antibodies to the Petition 870260065483, dated 03 / 07 / 2026, pp. 170 / 214 If antigen 160 / 195 is detected in mouse serum, the mouse spleen is collected and splenocytes are isolated. The splenocytes are then fused using well-known techniques to any suitable myeloma cells, for example, cells from the SP20 cell line available from the American Type Culture Collection (ATCC®) (Manassas, VA), to form hybridomas. Hybridomas are selected and cloned by limited dilution. In certain modalities, lymph nodes from immunized mice are collected and fused with NS0 myeloma cells.

[0297] The hybridoma cells thus prepared are seeded and proliferated in a suitable culture medium which preferably contains one or more substances that inhibit the proliferation or survival of unfused parental myeloma cells. For example, if the parental myeloma cells are devoid of the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the culture medium for hybridomas will typically include hypoxanthine, aminopterin and thymidine (HAT medium), such substances prevent the growth of HGPRT-deficient cells.

[0298] Specific modalities employ myeloma cells that fuse effectively, sustain stable high-level antibody production by selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these myeloma cell lines are murine myeloma cell lines, such as the NS0 cell line or those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, CA, USA, and SP-2 or X63-Ag8653 cells available from the American Type Culture Collection, Rockville, MD, USA. Mouse-human heteromyeloma and human myeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor D (1984) J Immunol 133: 3001-3005; Brodeur et al., Techniques and applications of monoclonal antibody production, pages 51-63 (Marcel Dekker, Inc., New York, 1987), each of which is incorporated herein by reference in its entirety. Petition 870260065483, dated 03 / 07 / 2026, pp. 171 / 214 161 / 195

[0299] The culture medium in which hybridoma cells are growing is analyzed for the production of monoclonal antibodies directed against TIM-3 (e.g., human TIM-3). The binding specificity of monoclonal antibodies produced by hybridoma cells is determined by methods known in the art, for example, immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA).

[0300] After identifying hybridoma cells that produce antibodies of the desired specificity, affinity, and / or activity, clones can be subcloned by limiting dilution procedures and proliferated by standard methods (Goding JW (Ed), Monoclonal Antibodies: Principles and Practice, supra). Suitable culture media for this purpose include, for example, D-MEM medium or RPMI 1640. In addition, hybridoma cells can be cultured in vivo as ascites tumors in an animal.

[0301] The monoclonal antibodies secreted by the subclones are adequately separated from the culture medium, ascites fluid or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis or affinity chromatography.

[0302] The antibodies described herein include antibody fragments that recognize specific TIM-3 (e.g., human TIM-3) and can be generated by any technique known to those skilled in the art. For example, the Fab and F(ab')2 fragments described herein can be produced recombinantly or by proteolytic cleavage of immunoglobulin molecules using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab')2 fragments). A Fab fragment corresponds to one of the two identical arms of an antibody molecule and contains the complete light chain paired with the VH and CHI domains of the heavy chain. An F(ab')2 fragment Petition 870260065483, dated 03 / 07 / 2026, pages 172 / 214 162 / 195 contains the two antigen-binding arms of an antibody molecule linked by disulfide bridges in the hinge region.

[0303] Furthermore, the antibodies described in this document can also be generated using various phage presentation methods known in the art. In phage presentation methods, functional antibody domains are displayed on the surface of phage particles carrying the polynucleotide sequences that encode them. In particular, the DNA sequences encoding the VH and VL domains are amplified from animal cDNA libraries (e.g., human or murine cDNA libraries from affected tissues). The DNA encoding the VH and VL domains is recombined together with an scFv ligand by PCR and cloned into a phagomid vector. The vector is electroporated onto E. coli, and the E. coli is infected with helper phage. The phage used in these methods is a typically filamentous phage including fd and M13, and the VH and VL domains are usually recombinantly fused to either phage gene III or gene VIII.The phage expressing an antigen-binding domain that binds to a particular antigen can be selected or identified with antigen, for example, using antigen-identified or antigen-bound or captured antigen on a solid surface or sphere. Examples of phage presentation methods that can be used to manufacture the antibodies described in this document include those disclosed in Brinkman U et al., (1995) J Immunol Methods 182: 41-50; Ames RS et al., (1995) J Immunol Methods 184: 177-186; Kettleborough CA et al., (1994) Eur J Immunol 24: 952-958; Persic L et al., (1997) Gene 187: 9-18; Burton DR & Barbas CF (1994) Advan Immunol 57: 191-280; PCT Application No. PCT / GB91 / 001134; International Publication Nos. WO 90 / 02809, WO 91 / 10737, WO 92 / 01047, WO 92 / 18619, WO 93 / 11236, WO 95 / 15982, WO 95 / 20401 and WO 97 / 13844; and US Patent Nos. 5,698,426, 5,223,409, 5,403,484, 5,580,717, 5,427,908, 5,750,753, 5,821,047, 5,571,698, 5,427,908. Petition 870260065483, dated 03 / 07 / 2026, pp. 173 / 214 163 / 195 5,516,637, 5,780,225, 5,658,727, 5,733,743 and 5,969,108, all of which are incorporated herein by reference in their entirety.

[0304] As described in the references above, after phage selection, the antibody-coding regions of the phage can be isolated and used to generate whole antibodies, including human antibodies or any other desired antigen-binding fragment, and expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, for example, as described below. Techniques for recombinantly producing antibody fragments such as Fab, Fab', and F(ab')2 fragments can also be employed using methods known in the art, such as those disclosed in PCT publication no. WO 92 / 22324; Mullinax RL et al., (1992) BioTechniques 12(6): 864-869; Sawai H et al., (1995) Am J Reprod Immunol 34: 26-34; and Better M et al., (1988) Science 240: 1041-1043, each of which is incorporated into the present document by reference in its entirety.

[0305] In certain embodiments, to generate whole antibodies, PCR primers including VH or VL nucleotide sequences, a restriction site, and a flanking sequence to protect the restriction site can be used to amplify the VH or VL sequences from a template, for example, scFv clones. Using sets of cloning procedures known to those skilled in the art, PCR-amplified VH domains can be cloned into vectors expressing a VH constant region, and PCR-amplified VL domains can be cloned into vectors expressing a VL constant region, for example, the human kappa or lambda constant regions. The VH and VL domains can also be cloned into a vector expressing the necessary constant regions.Heavy chain conversion vectors and light chain conversion vectors are then co-transfected into cell lines to generate stable or transient cell lines that express full-length antibodies. Petition 870260065483, dated 03 / 07 / 2026, pages 174 / 214 164 / 195 example, IgG, using sets of procedures known to those skilled in the art.

[0306] A chimeric antibody is a molecule in which different portions of the antibody are derived from different immunoglobulin molecules. For example, a chimeric antibody may contain a variable region of a mouse or rat monoclonal antibody fused to a constant region of a human antibody. Methods for producing chimeric antibodies are known in the art. See, for example, Morrison SL (1985) Science 229: 1202-1207; Oi VT & Morrison SL (1986) BioTechniques 4: 214-221; Gillies SD et al., (1989) J Immunol Methods 125: 191-202; and US patents nos. 5,807,715, 4,816,567, 4,816,397 and 6,331,415, all of which are incorporated herein by reference in their entirety.

[0307] A humanized antibody has the ability to bind to a predetermined antigen and comprises a framework region that substantially has the amino acid sequence of a human immunoglobulin and CDRs that substantially have the amino acid sequence of a non-human immunoglobulin (e.g., a murine immunoglobulin). In particular embodiments, a humanized antibody also comprises at least a portion of an immunoglobulin constant (Fc) region, typically that of a human immunoglobulin. The antibody may also include CHi, hinge, CH2, CH3, and CH4 regions of the heavy chain. A humanized antibody may be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA, and IgE, and any isotype, including IgG1, IgG2, IgG3, and IgG4.Humanized antibodies can be produced using a variety of known art procedures, including, but not limited to, CDR grafting (European Patent No. EP 239400; International Publication No. WO 91 / 09967; and US Patents Nos. 5,225,539, 5,530,101 and 5,585,089), inactivation or recapping (European P...

Claims

1. Isolated antibody that binds specifically to human TIM-3, the antibody CHARACTERIZED in that it comprises a variable region of the heavy chain comprising the amino acid sequence presented in SEQ ID NO: 28, and a variable region of the light chain comprising the amino acid sequence presented in SEQ ID NO:

46.

2. Isolated antibody according to claim 1, the antibody CHARACTERIZED in that it comprises: (a) a constant region of the heavy chain selected from the group consisting of human IgG1, IgG2, IgGa, IgG4, IgA1 and IgA2, wherein optionally the constant region of the heavy chain is: (i) a variant of a wild-type human IgG constant region of the heavy chain, wherein the variant human IgG constant region of the heavy chain binds to a human Fc gamma receptor with lower affinity than the wild-type human IgG constant region of the heavy chain binds to the human Fc gamma receptor, wherein optionally the human Fc gamma receptor is selected from the group consisting of FcyRI, FcyRII and FcyRIII; (ii) an IgG1 comprising an N297A mutation, numbered according to the EU numbering system, optionally comprising the amino acid sequence of SEQ ID NO: 72;(iii) an IgG1 comprising an N297Q mutation, numbered according to the EU numbering system; (iv) a non-fucosylated IgG1; and / or (v) an IgG4 comprising an S228P mutation, numbered according to the EU numbering system, optionally comprising the amino acid sequence of SEQ ID NO: 74, and / or (b) a constant region of the light chain selected from the group consisting of human IgGK and IgGX, wherein optionally the constant region of the light chain comprises the amino acid sequence of SEQ ID NO: 76.; 3. Isolated antibody, according to claim 1 or 2, CHARACTERIZED in that the heavy chain and the light chain, respectively, comprise the amino acid sequences presented in SEQ ID NOS: 61 and 69.

4. Isolated antibody, according to any one of claims 1 to 3, the antibody CHARACTERIZED in that: (a) it is a human antibody; (b) it is antagonistic to human TIM-3; (c) it inactivates, reduces or inhibits an activity of human TIM-3; (d) it inhibits the binding of human TIM-3 to phosphatidylserine; (e) it induces the production of IFNγ by peripheral blood mononuclear cells (PBMCs) stimulated with staphylococcal enterotoxin A (SEA); (f) it induces the production of IFNγ or TNFα by tumor-infiltrating lymphocytes (TILs) stimulated with anti-CD3 and anti-CD28 antibodies; and / or (g) it is conjugated to a cytotoxic agent, cytostatic agent, toxin, radionuclide or detectable marker.

5. Pharmaceutical composition CHARACTERIZED in that it comprises the antibody, as defined in any one of claims 1 to 4, and a pharmaceutically acceptable vehicle or excipient.

6. Use of the isolated antibody, as defined in any one of claims 1 to 4, or of the pharmaceutical composition, as defined in claim 5, CHARACTERIZED in that it is for preparing a medicament to: (a) increase the activation of T cells in response to an antigen in an individual; or (b) treat cancer in an individual.

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