Anti-TIGIT antibodies and methods of use

Abstract

The present invention provides anti-TIGIT (T cell immunoreceptor with Ig and ITIM domains) antibodies and methods of using the same.

Description

[0001] This application is a divisional application of the invention application with the application date of September 23, 2016, Chinese application number 201680055442.3, and invention name “Anti-TIGIT Antibodies and Methods of Use”.

[0002] Sequence Listing

[0003] This application contains a sequence listing, which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on September 22, 2016, is named 50474-120WO3_Sequence_Listing_9_22_16_ST25 and is 209,318 bytes in size. Field of the Invention

[0004] The present invention relates to anti-TIGIT (T cell immunoreceptor with Ig and ITIM domains) antibodies and methods of use thereof. Background of the Invention

[0005] Immune-related diseases (e.g., cancer) are manifestations or consequences of complex biological pathways that are critical in normal physiology for responding to trauma or injury, initiating repair from trauma or injury, and initiating innate and acquired defenses. Disease or pathology occurs when these normal physiological pathways cause additional trauma or injury, which is directly related to the intensity of the response (e.g., as a consequence of abnormal regulation or overstimulation) or as a reaction to themselves.

[0006] Although the occurrence of these diseases often involves multi-step pathways and often involves multiple different biological systems / pathways, intervention at key points in one or more of these pathways may have an ameliorative or therapeutic effect. Therapeutic intervention may occur by antagonizing harmful processes / pathways or stimulating beneficial processes / pathways.

[0007] Many immune-related diseases are known and have been extensively studied. Such diseases include cancer (neoplasia), immune-mediated inflammatory diseases, non-immune-mediated inflammatory diseases, infectious diseases, and immunodeficiency diseases.

[0008] T lymphocytes (T cells) are an important component of the mammalian immune response. T cells recognize antigens associated with self-molecules encoded by genes within the major histocompatibility complex (MHC). The antigen can be displayed on the surface of antigen presenting cells (APCs), virus-infected cells, cancer cells, transplants, etc. together with MHC molecules. The T cell system removes these cells that have changed and pose a health threat to the host mammal. T cells include helper T cells and cytotoxic T cells. Helper T cells proliferate in large quantities after recognizing the antigen-MHC complex on the APC. Helper T cells also secrete a variety of cytokines (i.e., lymphokines), which play a central role in the activation of B cells, cytotoxic T cells, and a variety of other cells involved in the immune response.

[0009] Regulatory T cells (Tregs) are a subset of helper T cells that play a crucial role in suppressing autoreactive immune responses and are often found in sites of chronic inflammation, such as tumor tissue. Tregs are phenotypically defined by high cell surface expression of CD25, CLTA4, GITR, and neuropilin-1 (NRP-1) and are under the control of the transcription factor FOXP3. Tregs exert their suppressive function on activated T cells through contact-dependent mechanisms and cytokine production. Tregs also regulate immune responses through direct interactions with ligands on dendritic cells (DCs), such as CD40L ligation and interaction of CTLA4 with B7 molecules on DCs, which trigger induction of indoleamine 2,3-dioxygenase (IDO). DCs are professional APCs capable of inducing immunity or tolerance to self- or non-self antigens. DC-expanded Tregs suppress alloreactive responses in vitro, and when adoptively transferred, appropriate Tregs suppress diabetes or experimentally induced asthma in NODscid mice. Specific interactions of DC ligands with Tregs can also abrogate their suppressive function, such as the engagement of GITR in mice, suggesting that DCs may have a moonlighting role in regulating Treg function.

[0010] The molecules CTLA4 and GITR are representatives of ligands defined within the CD28-B7 and TNF superfamilies of costimulatory / inhibitory molecules, respectively. These molecules are highly expressed on Tregs but are typically upregulated on activated T cells. More recently, a protein called TIGIT (for T cell immunoreceptors with Ig and ITIM domains) has been identified as a cell surface-bound protein that is specifically expressed in T cells and has an IgV domain, a transmembrane domain, and two putative immunoreceptor tyrosine inhibitory (ITIM) motifs. TIGIT is shown to be expressed particularly on Tregs and memory T cell subsets, as well as NK cells. Because there is an unmet need for new therapeutic agents and methods for treating immune-related disorders, particularly cancer, described herein are unexpectedly effective therapeutic compositions, such as anti-TIGIT antibodies and compositions thereof, and methods for treating immune-related disorders and cancers that involve regulating the interaction between TIGIT and its binding partners. SUMMARY OF THE INVENTION

[0011] The present invention provides anti-TIGIT (T cell immunoreceptor with Ig and ITIM domains) antibodies and variants thereof having improved properties, including, for example, binding affinity, cross-reactivity, pharmacokinetics, and / or expression. In particular, the present invention provides anti-TIGIT antibodies and variants thereof having, for example, high binding affinity to human TIGIT; cross-reactivity between human TIGIT, cynomolgus monkey (cyno) TIGIT, and / or rabbit TIGIT; desirable clearance properties in cynomolgus monkeys; and biochemical and biophysical properties that confer high stability to the antibodies and variants thereof.

[0012] In one aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody binds to an epitope on human TIGIT that comprises one or more of the amino acid residues Ser78, Ser80, and Lys82 of human TIGIT. In one embodiment, the epitope comprises amino acid residues Ser80 and Lys82 of human TIGIT. In another embodiment, the epitope comprises amino acid residues Ser78, Ser80, and Lys82 of human TIGIT. In another embodiment, the epitope further comprises amino acid residue Ala67 of human TIGIT. In another embodiment, the epitope further comprises one or more additional amino acid residues selected from the group consisting of Glu60, Leu65, and Ile68 of human TIGIT. In another embodiment, the epitope further comprises one or more additional amino acid residues selected from the group consisting of Gln56, Asn70, Leu73, and His111 of human TIGIT. In another embodiment, the epitope further comprises one or more additional amino acid residues selected from the group consisting of Thr55, Asn58, Asp63, Gln64, His76, Ile77, and Pro79 of human TIGIT. In another embodiment, the epitope consists of amino acid residues Thr55, Gln56, Asn58, Glu60, Asp63, Gln64, Leu65, Ala67, Ile68, Asn70, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT.

[0013] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody binds to an epitope on human TIGIT that comprises one or more of amino acid residues Thr55, Ser80, and Lys82 of human TIGIT. In one embodiment, the epitope comprises amino acid residue Lys82 of human TIGIT. In another embodiment, the epitope comprises amino acid residues Thr55, Ser80, and Lys82 of human TIGIT. In another embodiment, the epitope further comprises amino acid residue Gln56 of human TIGIT. In another embodiment, the epitope further comprises amino acid residues Ile77 or Pro79 of human TIGIT. In another embodiment, the epitope further comprises amino acid residues Ile77 and Pro79 of human TIGIT. In another embodiment, the epitope further comprises amino acid residues Asn58 or Glu60 of human TIGIT. In another embodiment, the epitope further comprises amino acid residues Asn58 and Glu60 of human TIGIT. In another embodiment, the epitope further comprises one or more additional amino acid residues selected from the group consisting of Leu65, Ile68, Leu73, His76, Ser78, and His111 of human TIGIT. In another embodiment, the epitope further comprises amino acid residues Leu65, Ile68, Leu73, His76, Ser78, and His111 of human TIGIT. In yet another embodiment, the epitope consists of Thr55, Gln56, Asn58, Glu60, Leu65, Ile68, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT.

[0014] In another aspect, the invention features an antibody that specifically binds human TIGIT, wherein the antibody comprises a paratope comprising one or more amino acid residues selected from the group consisting of heavy chain variable region amino acid residues Asn32, Tyr52, Arg52b, Phe53, Lys54, Tyr56, Asp58, Tyr99, Asp100, Leu100a, Leu100b, and Ala100c and light chain variable region amino acid residues Tyr27d, Tyr92, Ser93, Thr94, and Phe96. In one embodiment, the paratope consists of heavy chain variable region amino acid residues Asn32, Tyr52, Arg52b, Phe53, Lys54, Tyr56, Asp58, Tyr99, AsplOO, LeulOOa, LeulOOb, and Ala100c and light chain variable region amino acid residues Tyr27d, Tyr92, Ser93, Thr94, and Phe96. In any of the above aspects, the antibody may be capable of binding to rabbit TIGIT.

[0015] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody binds to an epitope on human TIGIT comprising one or more amino acid residues selected from the group consisting of Gln53, His111, and Tyr113 of human TIGIT. In some embodiments, the epitope further comprises Gln56 of human TIGIT. In some embodiments, the epitope further comprises Glu60, Leu65, Ile68, Asn70, Leu73, and His76 of human TIGIT.

[0016] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody comprises the following six hypervariable regions (HVRs): HVR-H1 comprising the amino acid sequence of SNSAAWN (SEQ ID NO: 1); HVR-H2 comprising the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO: 2); HVR-H3 comprising the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO: 3); HVR-L1 comprising the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO: 4); HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 5); and HVR-L3 comprising the amino acid sequence of QQYYSTPFT (SEQ ID NO: 6). In some embodiments, the antibody further comprises the following light chain variable region framework regions (FRs): FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7); FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8); FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10). In some embodiments, the antibody further comprises the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11), wherein X1 is Q or E; FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14). In some embodiments, the antibody further comprises the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 15); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14).In some embodiments, the antibody further comprises the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 16); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14). In some embodiments, the antibody is capable of binding to rabbit TIGIT.

[0017] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody comprises the following six HVRs: HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and HVR-L3 comprising the amino acid sequence of QHLHGYPX1N (SEQ ID NO: 22), wherein X1 is C or S. In some embodiments, the antibody comprises the following six HVRs: HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and HVR-L3 comprising the amino acid sequence of QHLHGYPSN (SEQ ID NO: 23). In some embodiments, the antibody further comprises the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 24); FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27).In some embodiments, the antibody comprises the following six HVRs: HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and HVR-L3 comprising the amino acid sequence of QHLHGYPCN (SEQ ID NO: 28). In some embodiments, the antibody further comprises the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 29); FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27). In some embodiments, the antibody further comprises the following light chain variable region FRs: FR-L1 comprising the amino acid sequence of DIQLTQSPTFLSASVGDRVTITC (SEQ ID NO: 30); FR-L2 comprising the amino acid sequence of WYQQNPGKAPKLLIY (SEQ ID NO: 31); FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEFTLTISSLQPEDFVTYYC (SEQ ID NO: 32); and FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 33).

[0018] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody comprises (a) a heavy chain variable region (VH) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 34 or 35; (b) a light chain variable region (VL) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36; or (c) a heavy chain variable region as in (a) and a light chain variable region as in (b). In some embodiments, the antibody comprises (a) a heavy chain variable region (VH) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 34; (b) a light chain variable region (VL) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36; or (c) a heavy chain variable region as in (a) and a light chain variable region as in (b). In some embodiments, the antibody comprises (a) a heavy chain variable region (VH) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 35; (b) a light chain variable region (VL) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36; or (c) a heavy chain variable region as in (a) and a light chain variable region as in (b). In some embodiments, the antibody is capable of binding to rabbit TIGIT.

[0019] In another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody comprises (a) a heavy chain variable region (VH) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 37; (b) a light chain variable region (VL) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 38; or (c) a heavy chain variable region as in (a) and a light chain variable region as in (b).

[0020] In yet another aspect, the invention features an antibody that specifically binds to human TIGIT, wherein the antibody comprises (a) a heavy chain variable region (VH) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 39; (b) a light chain variable region (VL) having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 40; or (c) a heavy chain variable region as in (a) and a light chain variable region as in (b).

[0021] In any of the aspects described above, the antibody may be able to bind to both human TIGIT and cynomolgus monkey (cyno) TIGIT, but not to mouse TIGIT. In some embodiments, the antibody binds to human TIGIT with a Kd of about 10 nM or less and binds to cynomolgus monkey TIGIT with a Kd of about 10 nM or less. In some embodiments, the antibody binds to human TIGIT with a Kd of about 0.1 nM to about 1 nM and binds to cynomolgus monkey TIGIT with a Kd of about 0.5 nM to about 1 nM. In some embodiments, the antibody binds to human TIGIT with a Kd of about 0.1 nM or less and binds to cynomolgus monkey TIGIT with a Kd of about 0.5 nM or less.

[0022] In any of the aspects described above, the antibody may be an antagonist antibody or an agonist antibody.

[0023] In some embodiments, the antagonist antibody specifically binds to TIGIT and inhibits or blocks the interaction of TIGIT with the poliovirus receptor (PVR). In some embodiments, the antagonist antibody inhibits intracellular signaling mediated by TIGIT binding to PVR. In some embodiments, the antagonist antibody inhibits or blocks human TIGIT binding to human PVR with an IC50 value of 10nM or less. In some embodiments, the antagonist antibody inhibits or blocks human TIGIT binding to human PVR with an IC50 value of 1nM to about 10nM. In some embodiments, the antagonist antibody inhibits or blocks cynomolgus monkey TIGIT binding to cynomolgus monkey PVR with an IC50 value of 50nM or less. In some embodiments, the antagonist antibody inhibits or blocks cynomolgus monkey TIGIT binding to cynomolgus monkey PVR with an IC50 value of 1nM to about 50nM. In some embodiments, the antagonist antibody inhibits or blocks cynomolgus monkey TIGIT binding to cynomolgus monkey PVR with an IC50 value of 1nM to about 5nM.

[0024] In some embodiments, the agonist antibody specifically binds to TIGIT and stimulates the interaction of PVR with CD226 or CD96. In some embodiments, the agonist antibody specifically binds to TIGIT and stimulates the interaction of PVR with CD226 and CD96. In some embodiments, the agonist antibody specifically binds to TIGIT and stimulates the interaction of human PVR with human CD226 and human CD96. In some embodiments, the agonist antibody specifically binds to TIGIT and stimulates the interaction of cynomolgus monkey PVR with cynomolgus monkey CD226 and cynomolgus monkey CD96.

[0025] In another aspect, the invention features an isolated antibody that competes for binding to TIGIT with the antibody of any of the above aspects.

[0026] In another aspect, the invention features an isolated antibody that binds to the same epitope as the antibody of any of the above aspects.

[0027] In some embodiments of any of the aspects described above, the antibody is monoclonal. In some embodiments, the antibody is human, humanized, or chimeric. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody has a clearance rate after administration (e.g., injection, e.g., intravenous injection) of less than about 10 ml / kg / day (e.g., about 3 ml / kg / day to about 10 ml / kg / day). In some embodiments, the antibody has a clearance rate of about 3 ml / kg / day to about 8 ml / kg / day. In some embodiments, the antibody is administered to a mammal (e.g., a monkey, such as a cynomolgus monkey, or a human). In some embodiments, the antibody is an antibody fragment that binds to TIGIT. In some embodiments, the antibody fragment is selected from the group consisting of Fab, Fab', Fab'-SH, Fv, single-chain variable fragment (scFv), and (Fab')2 fragments. In some embodiments, the antibody is an IgG class antibody. In some embodiments, the IgG class antibody is an IgG1 subclass antibody. In some embodiments, the antibodies described herein can be used as drugs. In some embodiments, the antibodies described herein can be used to treat cancer or delay cancer progression in a subject in need thereof. In some embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and hematological malignancies. In some embodiments, the antibodies described herein can be used to treat multiple myeloma (MM) or delay multiple myeloma (MM) progression. In some embodiments, the antibodies described herein can be used to treat immune-related diseases or delay immune-related disease progression in a subject in need thereof. In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.In some embodiments, the antibodies described herein can be used to enhance, strengthen, or stimulate an immune response or function in a subject in need thereof.

[0028] In another aspect, the invention features a polynucleotide (e.g., an isolated polynucleotide) encoding any antibody described herein. In another aspect, the invention features a vector (e.g., an expression vector) comprising a polynucleotide for expressing the antibody. In another aspect, the invention features a host cell comprising the aforementioned polynucleotide and / or vector. In some embodiments, the host cell is eukaryotic (e.g., a mammalian cell). In some embodiments, the eukaryotic cell is a 293 cell, a Chinese hamster ovary (CHO) cell, a yeast cell, or a plant cell. In some embodiments, the host cell is a prokaryotic cell. In some embodiments, the prokaryotic cell is an Escherichia coli.

[0029] In another aspect, the invention features a method for generating any antibody described herein, the method comprising cultivating a host cell comprising any of the aforementioned vectors (e.g., expression vectors) in a culture medium. In some embodiments, the method further comprises reclaiming the antibody from the host cell or the culture medium. In some embodiments, the host cell is a eukaryotic cell, such as a mammalian cell, such as a 293 cell, a Chinese hamster ovary (CHO) cell, a yeast cell, or a plant cell. In some embodiments, the host cell is a prokaryotic cell. In some embodiments, the prokaryotic cell is an Escherichia coli.

[0030] In another aspect, the invention features an immunoconjugate comprising any of the antibodies described herein and an agent (eg, a therapeutic agent, such as a cytotoxic agent).

[0031] In another aspect, the present invention features a composition comprising an antibody described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent. In some embodiments, the composition is used for diagnostic purposes (e.g., detecting TIGIT expression levels, such as TIGIT protein expression levels). In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the composition further comprises a PD-1 axis binding antagonist or another therapeutic agent.

[0032] In another aspect, the invention features the use of an antibody described herein in the manufacture of a medicament for treating cancer or delaying cancer progression in a subject in need thereof. In some embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and hematological malignancies. In some embodiments, the myeloma is MM. In some embodiments, the medicament is formulated for subcutaneous, intravenous, intramuscular, topical, oral, transdermal, intraperitoneal, intraorbital, by implant, by inhalation, intrathecally, intraventricularly, or intranasally. In some embodiments, the subject is human.

[0033] In another aspect, the invention features the use of the antibodies described herein in the manufacture of a medicament for treating an immune-related disease or delaying the progression of an immune-related disease in a subject in need thereof. In some embodiments, the immune-related disease is associated with a T-cell dysfunction disorder. In some embodiments, the T-cell dysfunction disorder is characterized by T-cell exhaustion. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity. In some embodiments, the medicament is formulated for subcutaneous, intravenous, intramuscular, topical, oral, transdermal, intraperitoneal, intraorbital, by implantation, by inhalation, intrathecal, intraventricular, or intranasal administration. In some embodiments, the subject is a human.

[0034] In another aspect, the invention features the use of an antibody described herein in the manufacture of a medicament for enhancing, increasing, or stimulating an immune response or function in a subject in need thereof. In some embodiments, the medicament is formulated for subcutaneous, intravenous, intramuscular, topical, oral, transdermal, intraperitoneal, intraorbital, by implant, by inhalation, intrathecally, intraventricularly, or intranasally administration. In some embodiments, the subject is human.

[0035] In another aspect, the invention features a method for treating cancer or delaying cancer progression in a subject, the method comprising administering to the subject an effective amount of any one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein, thereby treating the cancer or delaying cancer progression in the subject. In some embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic carcinoma, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and hematological malignancies. In some embodiments, the myeloma is MM.

[0036] In another aspect, the present invention features a method for treating an immune-related disease or delaying the progression of an immune-related disease in a subject, the method comprising administering to the subject an effective amount of one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein, thereby treating the immune-related disease or delaying the progression of the immune-related disease in the subject. In some embodiments, the immune-related disease is related to a T cell dysfunction disorder. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity.

[0037] In another aspect, the present invention features a method of increasing, enhancing, or stimulating an immune response or function in a subject, the method comprising administering to the subject an effective amount of one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein, thereby increasing, enhancing, or stimulating an immune response or function in the subject. In some embodiments, the method further comprises administering to the subject a PD-1 axis binding antagonist. In some embodiments, the PD-1 axis binding antagonist is administered before or after administration of the antibody. In some embodiments, the PD-1 axis binding antagonist is administered concurrently with the antibody. In some embodiments, the PD-1 axis binding antagonist is selected from the group consisting of a PD-1 binding antagonist, a PD-L1 binding antagonist, and a PD-L2 binding antagonist. In some embodiments, the PD-1 axis binding antagonist is a PD-1 binding antagonist. In some embodiments, the PD-1 binding antagonist inhibits PD-1 from binding to its ligand binding partner. In some embodiments, the PD-1 binding antagonist inhibits PD-1 from binding to PD-L1. In some embodiments, the PD-1 binding antagonist inhibits PD-1 from binding to PD-L2. In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In some embodiments, the PD-1 binding antagonist is selected from the group consisting of MDX 1106 (nivolumab), MK-3475 (pembrolizumab), CT-011 (pidilizumab), MEDI-0680 (AMP-514), PDR001, REGN2810, and BGB-108. In some embodiments, the PD-1 axis binding antagonist is a PD-L1 binding antagonist. In some embodiments, the PD-L1 binding antagonist inhibits PD-L1 from binding to PD-1. In some embodiments, the PD-L1 binding antagonist inhibits PD-L1 from binding to B7-1. In some embodiments, the PD-L1 binding antagonist inhibits PD-L1 from binding to both PD-1 and B7-1. In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of MPDL3280A (atezolizumab), YW243.55.S70, MDX-1105, MEDI4736 (durvalumab), and MSB0010718C (avelumab). In some embodiments, the antibody is MPDL3280A. In some embodiments, the PD-1 axis binding antagonist is a PD-L2 binding antagonist. In some embodiments, the PD-L2 binding antagonist is an anti-PD-L2 antibody. In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0038] In some embodiments, any of the treatment methods described above may further comprise administering an OX40 binding agonist to the subject. In some embodiments, the OX40 binding agonist is administered before or after administration of the antibody and / or the PD-1 axis binding antagonist. In some embodiments, the OX40 binding agonist is administered concurrently with the antibody and / or the PD-1 axis binding antagonist. In some embodiments, the OX40 binding agonist is selected from the group consisting of an OX40 agonistic antibody, an OX40L agonistic fragment, an OX40 oligomeric receptor, and an OX40 immunoadhesin. In some embodiments, the OX40 agonistic antibody depletes cells expressing human OX40. In some embodiments, the cells expressing human OX40 are CD4+ effector T cells. In some embodiments, the cells expressing human OX40 are regulatory T (Treg) cells. In some embodiments, the depletion is by ADCC and / or phagocytosis. In some embodiments, the OX40 agonistic antibody binds to human OX40 with an affinity of less than or equal to about 1 nM. In some embodiments, the OX40 agonist antibody binds to human OX40 with an affinity of less than or equal to about 0.45 nM. In some embodiments, the OX40 agonist antibody binds to human OX40 with an affinity of less than or equal to about 0.4 nM. In some embodiments, the OX40 agonist antibody binds to human OX40 with an EC50 of less than or equal to 0.3 μg / ml. In some embodiments, the OX40 agonist antibody binds to human OX40 with an EC50 of less than or equal to 0.2 μg / ml. In some embodiments, the OX40 agonist antibody increases CD4+ effector T cell proliferation and / or increases cytokine production by the CD4+ effector T cells compared to proliferation and / or cytokine production prior to treatment with the OX40 agonist antibody. In some embodiments, the OX40 agonist antibody increases memory T cell proliferation and / or cytokine production by memory T cells. In some embodiments, the cytokine production is IFN-γ production. In some embodiments, the OX40 agonist antibody inhibits Treg function. In some embodiments, the OX40 agonist antibody inhibits Treg suppression of effector T cell function (e.g., effector T cell proliferation and / or cytokine production). In some embodiments, the effector T cell is a CD4+ effector T cell. In some embodiments, the OX40 agonist antibody increases OX40 signaling in target cells expressing OX40. In some embodiments, the OX40 signaling is detected by monitoring NFkB downstream signaling. In some embodiments, the OX40 agonist antibody comprises a variant IgG1 Fc polypeptide comprising a mutation that eliminates binding to human effector cells and has reduced activity relative to an OX40 agonist antibody comprising a native sequence IgG1 Fc portion.In some embodiments, the OX40 agonist antibody comprises a variant IgG1 Fc polypeptide comprising a DANA mutation. In some embodiments, the OX40 agonist antibody comprises (a) a VH domain comprising (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 278, 279, or 280, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 281, 282, 283, 284, 285, or 286, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO: 287, 288, or 289; and (b) a VL domain comprising (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 290, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 291, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 292, 293, 294, 295, 296, 297, 298, or 299. In some embodiments, the OX40 agonist antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 278; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 281; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 287; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 290; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 291; and (f) HVR-L3 comprising an amino acid sequence selected from SEQ ID NO: 292. In some embodiments, the OX40 agonist antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 278; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 281; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 287; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 290; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 291; and (f) HVR-L3 comprising an amino acid sequence selected from SEQ ID NO: 297. In some embodiments, the OX40 agonist antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 278; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 281; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 287; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 290; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 291; and (f) HVR-L3 comprising an amino acid sequence selected from SEQ ID NO: 298.In some embodiments, the OX40 agonist antibody comprises a VH sequence that has at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 300-325. In some embodiments, the OX40 agonist antibody comprises a VH sequence that has at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 300. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 300. In some embodiments, the OX40 agonist antibody comprises a VH comprising one, two, or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 278, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 281, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 287. In some embodiments, the OX40 agonist antibody comprises a VL sequence that has at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 326-351. In some embodiments, the OX40 agonist antibody comprises a VL having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 326. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 326. In some embodiments, the OX40 agonist antibody comprises a VL comprising one, two, or three HVRs selected from: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 290; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 291; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 292. In some embodiments, the OX40 agonist antibody comprises (a) the VH sequence of SEQ ID NO: 300; (b) the VL sequence of SEQ ID NO: 326; or (c) a VH sequence as in (a) and a VL sequence as in (b). In some embodiments, the OX40 agonist antibody comprises (a) the VH sequence of SEQ ID NO: 319; (b) the VL sequence of SEQ ID NO: 345; or (c) the VH sequence as in (a) and the VL sequence as in (b). In some embodiments, the OX40 agonist antibody comprises (a) the VH sequence of SEQ ID NO: 320; (b) the VL sequence of SEQ ID NO: 346; or (c) the VH sequence as in (a) and the VL sequence as in (b). In some embodiments, the OX40 agonist antibody is antibody L106, antibody ACT35, MEDI6469, or MEDI0562. In some embodiments, the OX40 agonist antibody is a full-length IgG1 antibody.In some embodiments, the OX40 immunoadhesin is a trimeric OX40-Fc protein.

[0039] In some embodiments, any of the methods of treatment described above may further include administering to the subject an agent that reduces or inhibits one or more additional immune co-inhibitory receptors. In some embodiments, the one or more additional immune co-inhibitory receptors are selected from the group consisting of PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, and CD96. In some embodiments, any of the methods of treatment described above may further include administering to the subject an additional therapeutic agent. In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the antibody is administered subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. In some embodiments, the subject is human.

[0040] In another aspect, the invention features a kit comprising any one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein and a package insert containing instructions for using the antibodies to treat cancer or delay cancer progression in a subject. In some embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, small cell lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer, leukemia, lymphoma, myeloma, mycosis fungoides, Merkel cell carcinoma, and hematological malignancies. In some embodiments, the subject is human. In some embodiments, the myeloma is MM.

[0041] In another aspect, the invention features a kit comprising any one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein and a package insert comprising instructions for use of the antibody for treating an immune-related disease or delaying immune-related disease progression in a subject. In some embodiments, the immune-related disease is associated with a T cell dysfunction disorder. In some embodiments, the T cell dysfunction disorder is characterized by T cell exhaustion. In some embodiments, the immune-related disease is selected from the group consisting of unresolved acute infection, chronic infection, and tumor immunity. In some embodiments, the subject is a human.

[0042] In another aspect, the invention features a kit comprising any one or more (e.g., 1, 2, 3, 4, 5, or 6 or more) of the antibodies described herein and a package insert containing instructions for enhancing, increasing, or stimulating an immune response or function in a subject. In some embodiments, the subject is a human. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1A is a schematic diagram of the CHO-TIGIT binding assay.

[0044] Figure 1B It is a picture showing Figure 1A Depicted in Figure 5 are the results of a CHO-TIGIT binding assay using human TIGIT-expressing CHO cells and the indicated OMT-derived anti-TIGIT antibodies at various concentrations, as analyzed by FACS.

[0045] Figure 1C It is a picture showing Figure 1A Depicted are the results of a CHO-TIGIT binding assay using CHO cells expressing cynomolgus monkey TIGIT and the indicated OMT-derived anti-TIGIT antibodies at various concentrations, as analyzed by FACS.

[0046] Figure 1D It is a picture showing Figure 1A Depicted in Figure 5 are the results of a CHO-TIGIT binding assay using human TIGIT-expressing CHO cells and the indicated SD-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0047] Figure 1E It is a picture showing Figure 1A Depicted in Figure 5 are the results of a CHO-TIGIT binding assay using CHO cells expressing cynomolgus monkey TIGIT and the indicated SD-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0048] Figure 2A is a graph showing binding of the indicated OMT-derived anti-TIGIT antibodies to human CD4 T cells as a function of antibody concentration.

[0049] Figure 2B is a graph showing binding of the indicated OMT-derived anti-TIGIT antibodies to human CD8 T cells as a function of antibody concentration.

[0050] Figure 2C is a graph showing binding of the indicated SD-derived anti-TIGIT antibodies to human CD4 T cells as a function of antibody concentration.

[0051] Figure 2D is a graph showing binding of the indicated SD-derived anti-TIGIT antibodies to human CD8 T cells as a function of antibody concentration.

[0052] Figure 3A is a graph showing the results of a blocking ELISA assay as a function of antibody / Fab concentration using plates coated with human PVR-Fc fusion protein, human TIGIT, and the indicated anti-TIGIT antibodies (10A7 and 4.1D3.Q1E in Fab or IgG format) or an anti-CSF1 Fab control.

[0053] Figure 3B is a graph showing the results of a blocking ELISA assay as a function of antibody / Fab concentration using plates coated with cynomolgus PVR-Fc fusion protein, cynomolgus monkey TIGIT, and the indicated anti-TIGIT antibodies (10A7 and 4.1D3.Q1E in Fab or IgG format) or an anti-CSF1 Fab control.

[0054] Figure 3C is a graph showing the results of a blocking ELISA assay as a function of antibody / Fab concentration using plates coated with murine PVR-Fc fusion protein, murine TIGIT, and the indicated anti-TIGIT antibodies (10A7 and 4.1D3.Q1E in Fab or IgG format) or an anti-CSF1 Fab control.

[0055] Figure 3D Is a table showing Figure 3A-3C The results of the blocking ELISA assay are depicted in FIG.

[0056] Figure 4A Schematic diagram of the CHO-TIGIT PVR blocking assay.

[0057] Figure 4B It is a picture showing Figure 4A Depicted in Figure 5 are the results of a CHO-TIGIT PVR blocking assay using human TIGIT-expressing CHO cells and the indicated OMT-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0058] Figure 4C It is a picture showing Figure 4A Depicted in Figure 5 are the results of a CHO-TIGIT PVR blocking assay using CHO cells expressing cynomolgus monkey TIGIT and the indicated OMT-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0059] Figure 4D It is a picture showing Figure 4ADepicted in Figure 5 are the results of a CHO-TIGIT PVR blocking assay using human TIGIT-expressing CHO cells and the indicated SD-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0060] Figure 4E It is a picture showing Figure 4A Depicted in Figure 5 are the results of a CHO-TIGIT PVR blocking assay using CHO cells expressing cynomolgus monkey TIGIT and the indicated SD-derived anti-TIGIT antibodies at different concentrations, as analyzed by FACS.

[0061] Figure 5A is a graph showing the pharmacokinetic clearance of the indicated anti-TIGIT antibodies in cynomolgus monkeys following intravenous administration of 10 mg / kg as a function of serum concentration (μg / ml) over time (days).

[0062] Figure 5B Is a table showing Figure 5A Calculated clearance values ​​for each antibody tested in the pharmacokinetic clearance experiments in .

[0063] Figure 5C is a graph showing the pharmacokinetic clearance of anti-TIGIT antibodies h10A7.K4G3 and 4.1D3 in cynomolgus monkeys following 10 mg / kg intravenous administration as a function of serum concentration (μg / ml) over time (days; d0-d7).

[0064] Figure 5D Is a table showing Figure 5C Calculated clearance values ​​for the anti-TIGIT antibodies h10A7.K4G3 and 4.1D3 tested in the pharmacokinetic clearance experiments in FIG.

[0065] Figure 6A A perspective view of the crystal structure of 4.1D3 Fab bound to human TIGIT, with the 4.1D3 heavy chain (HC) and light chain (LC) regions labeled and color-coded.

[0066] Figure 6B A perspective view of the crystal structure of 4.1D3 Fab bound to human TIGIT superimposed with the known PVR-TIGIT structure (PDB 3UDW), showing that the 4.1D3 antibody and PVR have overlapping binding sites for TIGIT.

[0067] Figure 6C Figure 1 is a perspective view of the crystal structure of 4.1D3 Fab bound to human TIGIT, indicating the relative positioning of the PVR binding site. The relative positioning of the 72-79 loop of TIGIT is also indicated.

[0068] Figure 6D and Figure 6E are perspective views of the crystal structure of 4.1D3 Fab bound to human TIGIT from different angles, identifying certain key contact residues of 4.1D3 (paratope residues) and TIGIT (epitope residues). Figure 6E The boxed residues indicated in the figure were identified as functionally important epitope residues by alanine scanning mutagenesis.

[0069] Figure 7A is a graph showing the K of each identified anti-TIGIT antibody against the indicated human TIGIT alanine scanning mutants. D Value (nM).

[0070] Figure 7B is a graph showing the K of each identified anti-TIGIT antibody against the indicated human TIGIT alanine scanning mutants. D Values ​​(nM) are normalized to wild-type human TIGIT target. Alanine scanning mutants of TIGIT that result in a 1- to 10-fold or greater decrease in binding affinity are shaded in light grey and dark grey, respectively.

[0071] Figure 7C is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for 10A7 recognition by alanine scanning mutagenesis.

[0072] Figure 7D is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for h1A5 recognition by alanine scanning mutagenesis.

[0073] Figure 7E is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for 4.1D3 recognition by alanine scanning mutagenesis.

[0074] Figure 7F is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for 7.4A3 recognition by alanine scanning mutagenesis.

[0075] Figure 7G is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for 4.1A4 recognition by alanine scanning mutagenesis.

[0076] Figure 7H is a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for h6B2 recognition by alanine scanning mutagenesis.

[0077] Figure 7Iis a ribbon rendering of the human TIGIT structure, with spheres indicating residues identified as important for h7E7 recognition by alanine scanning mutagenesis.

[0078] Figure 8 A perspective view of the crystal structure of 1A5 Fab bound to human TIGIT, with the 1A5 heavy chain (HC) and light chain (LC) regions labeled and color-coded.

[0079] Figure 9 is a perspective view of the crystal structures of the 4.1D3-TIGIT, 1A5-TIGIT, and 10A7-TIGIT complexes, superimposed on each other with respect to TIGIT, showing that the three anti-TIGIT antibodies bind to human TIGIT at different epitopes.

[0080] Figure 10A is a perspective view of the crystal structure of 10A7 Fab bound to human TIGIT, with the 10A7 heavy chain (HC) and light chain (LC) regions labeled and color-coded.

[0081] Figure 10B Figure 1 is a perspective view of the crystal structure of 10A7 Fab bound to human TIGIT, indicating the relative positioning of the PVR binding site. The relative positioning of the 72-79 loop of TIGIT is also indicated.

[0082] Figure 11 A series of perspective views of the 4.1D3 (top left), 10A7 (center center), and 1A5 (bottom right) antibodies and the relative positioning of Ser78, Ser80, and Lys82 residues of human TIGIT. The structures indicate that Ser78, Ser80, and Lys82 residues of human TIGIT are key epitope residues for 4.1D3, but not for 10A7 or 1A5, where these three residues are located farther away from the antibody binding pocket.

[0083] Figure 12 is a series of graphs showing the expression of CD45, PD-1, TIGIT, and CD226 on CD4+ and CD8+ T cells isolated from the bone marrow of multiple myeloma (MM) patients (top row) and PD-1, TIGIT, and CD226 on CD4+ and CD8+ T cells isolated from the peripheral blood of healthy patients (bottom row), as assessed by multicolor flow cytometry.

[0084] Figure 13 A series of flow cytometry images (left) and accompanying images (right) show that TIGIT and PD-1 are co-expressed on CD4+ and CD8+ T cells in the bone marrow of MM patients. Detailed Description of the Invention

[0085] I. General Technology

[0086] The techniques and procedures described or referred to herein are generally well understood and commonly employed by those skilled in the art and are performed using conventional methods, such as, for example, the widely utilized methods described in Sambrook et al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (FM Ausubel et al., eds., (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GR Taylor, eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual, and Animal Cell Culture (RI Freshney, ed. (1987)); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Manual; and Notebook (JECellis, 1998) Academic Press; Animal Cell Culture (RIFreshney), 1987); Introduction to Cell and Tissue Culture (JPMather and PERoberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A.Doyle, JBGriffiths, and DG Newell, 1993-8) J.Wiley and Sons; Handbook of Experimental Immunology (eds. DMWeir and CC Blackwell); Gene Transfer Vectors for Mammalian Cells (JMMiller and MPEdited by Calos, 1987); PCR: The Polymerase Chain Reaction, (Edited by Mullis et al., 1994); Current Protocols in Immunology (Edited by J.E. Coligan et al., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (Edited by D. Catty., IRL Press, 1988 - 1989); Monoclonal Antibodies: A Practical Approach (Edited by P. Shepherd and C. Dean, Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (Edited by M. Zanetti and J.D. Capra, Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (Edited by V.T. DeVita et al., J.B. Lippincott Company, 1993).

[0087] II. Definitions

[0088] As used herein, the term "TIGIT" or "T cell immunoreceptor having Ig and ITIM domains" refers to any native TIGIT from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. TIGIT is also known in the art as DKFZp667A205, FLJ39873, protein 9 containing V cluster and immunoglobulin domains, protein 3 containing V cluster and transmembrane domains, VSIG9, VSTM3, and WUCAM. The term encompasses "full-length," unprocessed TIGIT (e.g., full-length human TIGIT having the amino acid sequence of SEQ ID NO: 352) as well as any form of TIGIT that results from processing in the cell (e.g., processed human TIGIT without a signal sequence having the amino acid sequence of SEQ ID NO: 353). The term also encompasses naturally occurring variants of TIGIT, such as splice variants or allelic variants. An exemplary amino acid sequence of human TIGIT can be found in UniProt Accession No. Q495A1.

[0089] The terms "anti-TIGIT antibody" and "antibody that specifically binds to TIGIT" refer to an antibody that can bind to TIGIT with sufficient affinity so that the antibody can be used as a diagnostic and / or therapeutic agent to target TIGIT. In one embodiment, the extent to which the anti-TIGIT antibody binds to unrelated, non-TIGIT proteins is less than about 10% of the binding of the antibody to TIGIT, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to TIGIT has a specific affinity of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8 M or less, such as 10 -8 M to 10 -13 M, for example 10 -9 M to 10 -13 In certain embodiments, the anti-TIGIT antibody binds to a TIGIT epitope that is conserved among TIGIT from different species or an epitope on TIGIT that allows cross-species reactivity, such as an epitope comprising amino acid residues Ser78, Ser80, and Lys82.

[0090] The term "antibody" includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibody compositions with multiple epitope specificities, multispecific antibodies (e.g., bispecific antibodies), diabodies, and single-chain molecules, and antibody fragments (e.g., Fab, F(ab')2, and Fv). The term "immunoglobulin" (Ig) is used interchangeably with "antibody" herein.

[0091] When used to describe the various antibodies disclosed herein, the term "isolated antibody" means an antibody that has been identified and separated and / or recovered from the cell or cell culture expressing it. The contaminating components of its natural environment refer to substances that typically interfere with the diagnosis or therapeutic use of the polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to a purity greater than 95% or 99%, as measured by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007). In preferred embodiments, the antibody is purified (1) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (2) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver stain. Isolated antibodies include antibodies in situ within recombinant cells, since at least one component of the polypeptide's natural environment will not be present. However, isolated polypeptides will generally be prepared by at least one purification step.

[0092] The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light chains (L) and two identical heavy chains (H). IgM antibodies are composed of 5 basic heterotetrameric units and an additional polypeptide called a J chain and contain 10 antigen binding sites, while IgA antibodies contain 2-5 basic 4-chain units that can polymerize to form multivalent assemblies in combination with J chains. In the case of IgG, the 4-chain unit is usually about 150,000 daltons. Each light chain is linked to the heavy chain by a covalent disulfide bond, while the two heavy chains are linked to each other by one or more disulfide bonds, the number of which depends on the isotype of the heavy chain. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (V) at the N-terminus. H ), followed by three (for α and γ chains) or four (for μ and ε isoforms) constant domains (C H Each light chain has a variable domain (V L ), followed by a constant domain at the other end (C L ). V L With V H Arranged together, and C L The first constant domain of the heavy chain (C H 1) Arranged together. It is believed that specific amino acid residues form an interface between the light chain and heavy chain variable domains. A V H and a V LPaired together to form an antigen binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and chapter 6. Based on the amino acid sequence of its constant domain, L chains from any vertebrate species can be classified into one of two distinct types, called kappa (κ) and lambda (λ). Based on the amino acid sequence of its heavy chain constant domain (C H ) amino acid sequence, immunoglobulins can be classified into different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated α, δ, ε, γ, and μ, respectively. H Due to minor differences in sequence and function, the gamma and alpha classes can be further divided into subclasses, for example humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0093] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of an antibody heavy or light chain. The heavy chain variable domain and light chain variable domain may be referred to as "VH" and "VL," respectively. These domains are generally the most variable parts of an antibody (relative to other antibodies of the same class) and contain the antigen-binding site.

[0094] The term "variable" refers to the fact that certain segments of the variable domain differ widely in sequence between antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not evenly distributed across the entire span of the variable domain. Instead, it is concentrated in three segments called hypervariable regions (HVRs) in both the light and heavy chain variable domains. The more highly conserved portions of the variable domain are called framework regions (FRs). The variable domains of native heavy and light chains each contain four FR regions, which mostly adopt a β-pleated sheet conformation and are connected by three HVRs that form loops connecting and, in some cases, forming part of the β-pleated sheet structure. The HVRs in each chain are held together in close proximity by the FR regions and, together with the HVRs of the other chain, contribute to the formation of the antibody's antigen-binding site (see Kabat et al., Sequences of Immunological Interest, 5th ed., National Institute of Health, Bethesda, MD. (1991)). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity.

[0095] A "blocking antibody" or "antagonist antibody" is an antibody that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, a blocking antibody or antagonist antibody substantially or completely inhibits the biological activity of the antigen. The anti-TIGIT antibodies of the present invention can block signaling through PVR, PVRL2, and / or PVRL3, thereby restoring the functional response of T cells (e.g., proliferation, cytokine production, target cell killing) from a dysfunctional state in response to antigen stimulation.

[0096] "Agonistic antibodies" or "activating antibodies" are antibodies that enhance or initiate signaling through the antigen to which they bind. In some embodiments, agonistic antibodies cause or activate signaling in the absence of a natural ligand. The OX40 agonistic antibodies of the present invention can increase memory T cell proliferation, increase cytokine production by memory T cells, inhibit Treg cell function, and / or inhibit Treg cell suppression of effector T cell function (such as effector T cell proliferation and / or cytokine production).

[0097] An "epitope" is a portion of an antigen to which an antibody specifically binds. For polypeptide antigens, an epitope is generally a peptide portion of about 4-15 amino acid residues, which may be continuous or discontinuous.

[0098] An "antibody that binds to the same epitope as a reference antibody" refers to an antibody that blocks the binding of the reference antibody to its antigen by 50% or more in a competition assay, and conversely, the reference antibody blocks the binding of the antibody to its antigen by 50% or more in a competition assay. Exemplary competition assays are provided herein.

[0099] "Paratope" means the portion of an antibody that selectively binds to an antigenic epitope. The paratope typically includes amino acids from the VH and VL chains of an antibody, particularly from the HVR regions of an antibody.

[0100] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical, except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerizations, amidations) that may be present in very small amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Unlike typical polyclonal antibody preparations that contain different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. Beyond their specificity, the advantage of monoclonal antibodies is that they are synthesized by hybridoma cultures and are not contaminated by other immunoglobulins. The modifier "monoclonal" indicates the characteristic of the antibody being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring the antibody to be generated by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention can be produced by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein., Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14(3):253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567), phage display technology (see, e.g., Clackson et al., Nature 352:624-628 (1991); Marks et al., J. Mol. Biol. 222:581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2):299-310 (2004); Lee et al., J. Mol. Biol. 340(5):1073-1093 (2004); Fellouse, Proc. Nat. Acad. Sci. USA 101(34):12467-12472 (2004); Lee et al., J. Immunol. Methods 284(1-2):119-132 (2004)), and techniques for producing human or human-like antibodies in animals that have part or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc.Natl.Acad.Sci.USA 90:2551 (1993); Jakobovits et al., Nature 362:255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016; Marks et al., Bio / Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-813 (1994); Fishwild et al., Nature Biotechnol. 14:845-851 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); Lonberg and Huszar, Intern. Rev. Immunol. 13:65-93 (1995)). .

[0101] The term "naked antibody" refers to an antibody that is not conjugated to a cytotoxic moiety or a radiolabel.

[0102] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably to refer to an antibody in its substantially complete form, as distinguished from an antibody fragment. Specifically, intact antibodies include those having heavy and light chains (including an Fc region). The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, intact antibodies may possess one or more effector functions.

[0103] "Antibody fragments" comprise a portion of an intact antibody, preferably the antigen-binding and / or variable regions of an intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (see U.S. Patent 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10):1057-1062 (1995)); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, and a residual "Fc" fragment, the name of which reflects its ability to crystallize readily. The Fab fragment consists of an intact L chain and the variable region (V H ) and the first constant domain of the heavy chain (C H1) Composition. Each Fab fragment is monovalent for antigen binding, that is, it has one antigen binding site. Pepsin treatment of the antibody produces a larger F(ab')2 fragment, which is roughly equivalent to two Fab fragments connected by disulfide bonds, with different antigen binding activities and still able to cross-link antigen. The Fab' fragment is due to the C H Fab fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the constant domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine ​​residues in the constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0104] The Fc fragment contains the carboxyl-terminal portions of two H chains held together by disulfide bonds. The effector functions of an antibody are determined by sequences in the Fc region, which is also recognized by Fc receptors (FcRs) found on certain cell types.

[0105] "Fv" is the smallest antibody fragment that contains a complete antigen recognition and binding site. This fragment consists of a dimer of one heavy-chain variable region domain and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains arise six hypervariable loops (three loops each from the heavy and light chains) that contribute the amino acid residues required for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only the three HVRs specific for an antigen) has the ability to recognize and bind antigen, albeit with lower affinity than the entire binding site.

[0106] "Single-chain Fv", also abbreviated as "sFv" or "scFv", is a Fv that contains V H and V L Preferably, the sFv polypeptide is in the V H With V L The domains further comprise polypeptide linkers that enable the sFv to form the desired structure for antigen binding. For a review of sFv, see Pluckthun, in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., Springer-Verlag, New York, pp. 269-315, 1994.

[0107] The "functional fragment" of an antibody of the present invention comprises a portion of an intact antibody, generally including the antigen-binding or variable region of the intact antibody or an Fc region of an antibody that retains or has improved FcR binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0108] The term "diabody" refers to a H and V L Small antibody fragments prepared by constructing sFv fragments (see previous paragraph) using short linkers (about 5-10 residues) between domains. Due to the short linker, the V domains perform interchain rather than intrachain pairing, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. Bispecific diabodies are heterodimers of two "crossover" sFv fragments, in which the V domains of the two antibodies are H and V L The domains are present on different polypeptide chains. Diabodies are described in more detail in, for example, EP 404,097; WO 93 / 11161; Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993).

[0109] Monoclonal antibodies are specifically included herein as "chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired biological activity (U.S. Patent No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA 81: 6851-6855 (1984)). Chimeric antibodies of interest herein include "primatized" antibodies in which the antigen-binding region of the antibody is derived from an antibody generated by immunizing a macaque monkey with, for example, an antigen of interest. As used herein, "humanized antibodies" are a subset of "chimeric antibodies".

[0110] "Humanized" forms of non-human (e.g., mouse) antibodies refer to chimeric antibodies that minimally comprise sequences derived from non-human immunoglobulins. In one embodiment, a humanized antibody refers to an immunoglobulin (immunoglobulin) in which the HVR residues (defined below) in the human immunoglobulin (recipient antibody) are replaced with HVR residues from a non-human species (donor antibody) (such as mouse, rat, rabbit, or non-human primate) with desired specificity, affinity, and / or ability. In some cases, the framework ("FR") residues of the human immunoglobulin are replaced with corresponding non-human residues. In addition, humanized antibodies may be included in residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further improve the performance of the antibody, such as binding affinity. In general, a humanized antibody will comprise at least one, typically two, substantially all variable domains, in which all or substantially all hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or substantially all FR regions are those of a human immunoglobulin sequence, although the FR regions may contain one or more individual FR residue substitutions that improve antibody performance (such as binding affinity, isomerization, immunogenicity, etc.). The number of these amino acid substitutions in the FRs is generally no more than 6 in the H chain and no more than 3 in the L chain. The humanized antibody will optionally also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For more details, see, for example, Jones et al., Nature 321: 522-525 (1986); Riechmann et al., Nature 332: 323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2: 593-596 (1992). See also, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1: 105-115 (1998); Harris, Biochem. Soc. Transactions 23: 1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5: 428-433 (1994); and U.S. Pat. Nos. 6,982,321 and 7,087,409.

[0111] "Human antibody" refers to an antibody that has an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human and / or is produced using any of the techniques disclosed herein for producing human antibodies. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol. 227: 381 (1991); Marks et al., J. Mol. Biol. 222: 581 (1991)). Also useful for preparing human monoclonal antibodies are the methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol. 147 (1): 86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering an antigen to a transgenic animal, such as an immunized xenogeneic mouse, that has been modified to produce human antibodies in response to antigenic stimulation but whose endogenous loci have been disabled (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584, regarding XENOMOUSE). TM See also, e.g., Li et al., Proc. Natl. Ascad. Sci. USA, 103:3557-3562 (2006), for human antibodies generated via human B-cell hybridoma technology.

[0112] The term "hypervariable region", "HVR" or "HV" as used herein refers to a region in an antibody variable domain that is highly variable in sequence and / or forms a structurally defined loop. Typically, an antibody comprises six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). In natural antibodies, H3 and L3 show the greatest diversity of these six HVRs, and it is believed that H3 in particular plays a unique role in conferring precise specificity on antibodies. See, for example, Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in: Methods in Molecular Biology 248:1-25 (Lo ed., Human Press, Totowa, NJ, 2003). In fact, naturally occurring camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains. See, eg, Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0113] Many HVR descriptions are used and covered herein. The Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia refers instead to the positions of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and the Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software. The "contact" HVRs are based on analysis of available complex crystal structures. The residues in each of these HVRs are recorded below.

[0114]

[0115] HVRs may include "extended HVRs" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH. For each of these definitions, the variable domain residues are numbered according to Kabat et al., supra.

[0116] The expression "variable domain residue numbering according to Kabat" or "amino acid position numbering according to Kabat" and variations thereof refer to the numbering system used by Kabat et al., supra, for editing of antibody heavy chain variable domains or light chain variable domains. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening or insertion of a variable domain FR or HVR. For example, the heavy chain variable domain may contain a single amino acid insertion after H2 residue 52 (residue 52a according to Kabat) and inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat residue numbering of a given antibody can be determined by aligning the antibody sequence with the "standard" Kabat numbering sequence for regions of homology.

[0117] "Framework" or "FR" residues are those variable domain residues other than the HVR residues as herein defined.

[0118] A "human consensus framework" or "acceptor human framework" refers to a framework that represents the most common amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Typically, the selection of human immunoglobulin VL or VH sequences is from a subset of variable domain sequences. Typically, the sequence subset is as described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991). Examples include, for VL, the subgroup can be subgroup κI, κII, κIII or κIV, as in Kabat et al., supra. Additionally, for VH, the subgroup can be subgroup I, subgroup II, or subgroup III, as in Kabat et al., supra. Alternatively, a human consensus framework can be derived from the above frameworks, wherein specific residues, such as human framework residues, are selected based on their homology to the donor framework by aligning the donor framework sequence with a collection of various human framework sequences. The acceptor human framework or human consensus framework "derived" from a human immunoglobulin framework can comprise the same amino acid sequence thereof, or it can contain pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less.

[0119] A "VH subgroup III consensus framework" comprises a consensus sequence obtained from the amino acid sequences in variable heavy chain subgroup III of Kabat et al., supra. In one embodiment, the VH subgroup III consensus framework amino acid sequence comprises at least a portion or all of each of the following sequences: EVQLVESGGGLVQPGGSLRLSCAAS (HC-FR1) (SEQ ID NO: 229), WVRQAPGKGLEWV (HC-FR2) (SEQ ID NO: 230), RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (HC-FR3) (SEQ ID NO: 232), and WGQGTLVTVSA (HC-FR4) (SEQ ID NO: 232).

[0120] The "VLκI consensus framework" comprises a consensus sequence obtained from the amino acid sequences in variable light chain kappa subgroup I of Kabat et al., supra. In one embodiment, the VL subgroup I consensus framework amino acid sequence comprises at least a portion or all of each of the following sequences: DIQMTQSPSSLSASVGDRVTITC (LC-FR1) (SEQ ID NO: 233), WYQQKPGKAPKLLIY (LC-FR2) (SEQ ID NO: 234), GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (LC-FR3) (SEQ ID NO: 235), and FGQGTKVEIKR (LC-FR4) (SEQ ID NO: 236).

[0121] "Amino acid modification" at a designated position (e.g., an Fc region) refers to the replacement or deletion of a designated residue, or the insertion of at least one amino acid residue near a designated residue. Insertion "near" a designated residue means insertion within one to two residues thereof. Insertion can be at the N-terminus or C-terminus of a designated residue. Preferred amino acid modifications herein are substitutions.

[0122] "Affinity" refers to the strength of the sum of all non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen, such as TIGIT). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed using a dissociation constant (Kd). Affinity can be measured by conventional methods known in the art, including methods described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0123] An "affinity matured" antibody is one that has one or more alterations in one or more HVRs of the antibody that result in an improvement in the affinity of the antibody for the antigen compared to a parent antibody that does not have those alterations. In one embodiment, affinity matured antibodies have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies can be generated by procedures known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describe affinity maturation by VH and VL domain shuffling. Random mutagenesis of HVR and / or framework residues is described in, for example, Barbas et al., Proc. Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al., Gene 169:147-155 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); Hawkins et al., J. Mol. Biol. 226:889-896 (1992).

[0124] As used herein, the terms "bind," "specifically bind," or "specific for" refer to a measurable and reproducible interaction, such as binding between a target and an antibody, that determines the presence of the target in the presence of a heterogeneous population of molecules (including biological molecules). For example, an antibody that specifically binds to a target (which may be an epitope) is an antibody that binds to this target with greater affinity, avidity, more readily, and / or for a greater duration than it binds to other targets. In one embodiment, the extent to which the antibody binds to an unrelated target is less than about 10% of the extent to which the antibody binds to the target, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that specifically binds to a target has a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. In certain embodiments, the antibody specifically binds to an epitope on a protein that is conserved between proteins from different species. In another embodiment, specific binding may include but does not require exclusive binding. As used herein, the term may, for example, be represented by a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. -4 M or lower, or 10 -5 M or lower, or 10 -6 M or lower, or 10 -7 M or lower, or 10 -8 M or lower, or 10 -9 M or lower, or 10 -10 M or lower, or 10 -11 M or lower, or 10 -12 M or lower for the target Kd or 10 -4 M to 10 -6 M or 10 -6 M to 10 -10 M or 10 -7 M to 10 -9 As will be appreciated by those skilled in the art, affinity and Kd values ​​are inversely correlated. High affinity for an antigen is measured by a low Kd value. In one embodiment, the term "specific binding" refers to binding whereby a molecule binds to a specific polypeptide or an epitope on a specific polypeptide without substantially binding to any other polypeptide or polypeptide epitope.

[0125] As used herein, the term "immunoadhesin" refers to an antibody-like molecule that combines the binding specificity of a heterologous protein ("adhesin") with the effector function of an immunoglobulin constant domain. Structurally, an immunoadhesin comprises a fusion of an amino acid sequence having a desired binding specificity different from the antigen recognition and binding site of an antibody (i.e., "heterologous") and an immunoglobulin constant domain sequence. The adhesin portion of an immunoadhesin molecule is typically a continuous amino acid sequence that comprises at least the binding site of a receptor or ligand. The immunoglobulin constant domain sequence in an immunoadhesin can be obtained from any immunoglobulin, such as IgG-1, IgG-2 (including IgG2A and IgG2B), IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE, IgD or IgM. Preferably, the Ig fusion comprises a substitution of at least one variable region in the Ig molecule with a domain of a polypeptide or antibody as described herein. In a particularly preferred embodiment, the immunoglobulin fusion comprises the hinge, CH2 and CH3, or the hinge, CH1, CH2 and CH3 regions, of an IgG1 molecule. For the generation of immunoglobulin fusions, see also U.S. Patent No. 5,428,130, issued June 27, 1995. For example, useful immunoadhesins for use in the combination therapies herein include polypeptides comprising an extracellular or OX40 binding portion of OX40L or an extracellular or OX40L binding portion of OX40 fused to a constant domain of an immunoglobulin sequence, such as OX40 ECD-Fc or OX40L ECD-Fc. Immunoadhesin combinations of Ig Fc and cell surface receptor ECD are sometimes referred to as soluble receptors.

[0126] "Fusion protein" and "fusion polypeptide" refer to a polypeptide having two covalently linked parts, wherein each part is a polypeptide with different properties. The property can be a biological property, such as in vitro or in vivo activity. The property can also be a simple chemical or physical property, such as binding to a target molecule, catalysis of a reaction, etc. The two parts can be directly linked by a single peptide bond, or linked via a peptide linker, but in-frame with each other.

[0127] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Although the boundaries of an immunoglobulin heavy chain Fc region can vary, a human IgG heavy chain Fc region is generally defined as the segment from the amino acid residue at its Cys226 or Pro230 position to the carboxyl terminus. The C-terminal lysine (residue 447, according to the EU numbering system) in the Fc region can be eliminated, for example, during the production or purification of the antibody, or by recombinant engineering of the nucleic acid encoding the antibody heavy chain. Thus, a composition of a complete antibody can include antibody groups in which all K447 residues have been eliminated, antibody groups in which no K447 residue has been eliminated, and antibody groups in which antibodies with and without K447 residues are mixed. Suitable native sequence Fc regions for use in the antibodies of the present invention include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4.

[0128] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. A preferred FcR is a native sequence human FcR. Furthermore, a preferred FcR is an FcR that binds to IgG antibodies (gamma receptors), including receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptors") and FcγRIIB ("inhibiting receptors"), which have similar amino acid sequences and differ primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see M. Annu. Rev. Immunol. 15:203-234 (1997). For review of FcRs, see Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991); Capel et al., Immunomethods 4:25-34 (1994); de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). The term "FcR" herein encompasses additional FcRs, including those identified in the future.

[0129] "Human effector cells" refer to leukocytes that express one or more FcRs and perform effector functions. In certain embodiments, the cells express at least FcγRIII and perform ADCC effector functions. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils. Effector cells can be isolated from their natural sources, such as blood.

[0130] "Effector functions" refer to those biological activities attributable to the Fc region of an antibody and vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[0131] The phrase "substantially reduced" or "substantially different" as used herein refers to a sufficiently high degree of difference between two values ​​(typically one associated with a molecule and the other associated with a reference / comparator molecule) that one skilled in the art would consider the difference between the two values ​​to be statistically significant within the context of the biological property being measured by the value (e.g., Kd value). For example, the difference between the two values ​​is greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, and / or greater than about 50% as a function of the value of the reference / comparator molecule.

[0132] The phrase "substantially similar" or "substantially identical" as used herein refers to a sufficiently high degree of similarity between two values ​​(e.g., one relating to an antibody of the invention and the other relating to a reference / comparator antibody) that one skilled in the art would consider the difference between the two values ​​to be of little or no biological and / or statistical significance within the context of the biological property measured by the value (e.g., Kd value). The difference between the two values ​​as a function of the reference / comparator value is, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and / or less than about 10%.

[0133] The term "antagonist" is used in the broadest sense to include any molecule that partially or completely blocks, inhibits, or neutralizes a biological activity of a native polypeptide disclosed herein. Similarly, the term "agonist" is used in the broadest sense to include any molecule that mimics a biological activity of a native polypeptide disclosed herein. Suitable agonist or antagonist molecules specifically include agonistic or antagonistic antibodies or antibody fragments, fragments or amino acid sequence variants of a native polypeptide, peptides, antisense oligonucleotides, small organic molecules, and the like. Methods for identifying agonists or antagonists of a polypeptide may comprise contacting the polypeptide with a candidate agonist or antagonist molecule and measuring a detectable change in one or more biological activities typically associated with the polypeptide.

[0134] As used herein, an "antagonistic antibody" to a target molecule refers to an antibody that interferes with the normal function of the target molecule, either by reducing the transcription or translation of the nucleic acid encoding the target molecule, or by inhibiting or blocking the activity of the target molecule, or both. For example, an antagonistic antibody to TIGIT may be considered to be an antibody or an anti-TIGIT binding fragment thereof that interferes with the normal function of TIGIT, either by reducing the transcription or translation of the nucleic acid encoding TIGIT, or by inhibiting or blocking the activity of the TIGIT polypeptide, or both. One of ordinary skill in the art will understand that in some cases, an antagonistic antibody to TIGIT may antagonize one TIGIT activity without affecting another TIGIT activity. For example, a desired antagonistic antibody to TIGIT for use in certain methods herein is an antagonistic antibody to TIGIT that antagonizes TIGIT activity in response to one of a PVR interaction, a PVRL3 interaction, or a PVRL2 interaction, e.g., does not affect or minimally affects any other TIGIT interaction.

[0135] The terms "TIGIT" antagonist and "antagonist of TIGIT activity or TIGIT expression" are used interchangeably and refer to a compound that interferes with the normal functioning of TIGIT by reducing the transcription or translation of TIGIT encoding nucleic acid, or by inhibiting or blocking the activity of a TIGIT polypeptide, or both. Examples of TIGIT antagonists include, but are not limited to, antisense polynucleotides, interfering RNA, catalytic RNA, RNA-DNA chimeras, TIGIT-specific aptamers, anti-TIGIT antibodies, TIGIT-binding fragments of anti-TIGIT antibodies, TIGIT-binding small molecules, TIGIT-binding peptides, and other polypeptides that specifically bind to TIGIT (including but not limited to TIGIT-binding fragments of one or more TIGIT ligands, optionally fused to one or more additional domains) such that the interaction between the TIGIT antagonist and TIGIT results in a reduction or cessation of TIGIT activity or expression. One of ordinary skill in the art will appreciate that in some cases, a TIGIT antagonist may antagonize one TIGIT activity without affecting the activity of another. For example, desirable TIGIT antagonists for use in certain methods herein are TIGIT antagonists that antagonize TIGIT activity in response to, for example, one of a PVR interaction, a PVRL3 interaction, or a PVRL2 interaction, but do not affect or minimally affect any other TIGIT interaction.

[0136] The terms "PVR antagonist" and "antagonist of PVR activity or PVR expression" are used interchangeably and refer to a compound that interferes with the normal function of PVR by reducing the transcription or translation of PVR-encoding nucleic acid, or by inhibiting or blocking the activity of a PVR polypeptide, or both. Examples of PVR antagonists include, but are not limited to, antisense polynucleotides, interfering RNA, catalytic RNA, RNA-DNA chimeras, PVR-specific aptamers, anti-PVR antibodies, PVR-binding fragments of anti-PVR antibodies, PVR-binding small molecules, PVR-binding peptides, and other polypeptides that specifically bind to PVR (including, but not limited to, PVR-binding fragments of one or more PVR ligands, optionally fused to one or more additional domains) such that the interaction between the PVR antagonist and PVR results in a reduction or cessation of PVR activity or expression. One of ordinary skill in the art will appreciate that in some cases, a PVR antagonist may antagonize one PVR activity without affecting another PVR activity. For example, desirable PVR antagonists for use in certain methods herein are PVR antagonists that antagonize PVR activity in response to TIGIT interaction but do not affect PVR-CD96 and / or PVR-CD226 interactions.

[0137] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners, thereby removing T cell dysfunction resulting from signaling along the PD-1 signaling axis—one result is restoration or enhancement of T cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0138] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with signal transduction derived from the interaction of PD-1 with one or more of its binding partners (such as PD-L1, PD-L2). In some embodiments, the PD-1 binding antagonist is a molecule that inhibits PD-1 from binding to one or more of its binding partners. In a specific aspect, the PD-1 binding antagonist inhibits PD-1 from binding to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with signal transduction derived from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative co-stimulatory signals mediated by or via cell surface proteins expressed on T lymphocytes (via PD-1 mediated signaling), thereby making dysfunctional T cells less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab) described herein. In another specific aspect, the PD-1 binding antagonist is MK-3475 (lambrolizumab) described herein. In another specific aspect, the PD-1 binding antagonist is CT-011 (pidilizumab) described herein. In another specific aspect, the PD-1 binding antagonist is MEDI-0680 (AMP-514) described herein. In another specific aspect, the PD-1 binding antagonist is PDR001 described herein. In another specific aspect, the PD-1 binding antagonist is REGN2810 described herein. In another specific aspect, the PD-1 binding antagonist is BGB-108 described herein.

[0139] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with signal transduction derived from the interaction of PD-L1 with one or more of its binding partners (such as PD-1, B7-1). In some embodiments, the PD-L1 binding antagonist is a molecule that inhibits PD-L1 from binding to its binding partner. In a specific aspect, the PD-L1 binding antagonist inhibits PD-L1 from binding to PD-1 and / or B7-1. In some embodiments, the PD-L1 binding antagonist includes anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with signal transduction derived from the interaction of PD-L1 with one or more of its binding partners (such as PD-1, B7-1). In one embodiment, the PD-L1 binding antagonist reduces negative co-stimulatory signals mediated by or via cell surface proteins expressed on T lymphocytes (via PD-L1 mediated signaling), thereby making dysfunctional T cells less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In a specific aspect, the anti-PD-L1 antibody is YW243.55.S70 described herein. In another specific aspect, the anti-PD-L1 antibody is MDX-1105 described herein. In yet another specific aspect, the anti-PD-L1 antibody is MPDL3280A (atezolizumab) described herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736 (durvalumab) described herein. In yet another specific aspect, the anti-PD-L1 antibody is YW243.55.S70 described herein. In yet another specific aspect, the anti-PD-L1 antibody is MSB0010718C (avelumab) described herein.

[0140] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with signal transduction derived from the interaction of PD-L2 with one or more of its binding partners (such as PD-1). In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits PD-L2 from binding to one or more of its binding partners. In a specific aspect, a PD-L2 binding antagonist inhibits PD-L2 from binding to PD-1. In some embodiments, PD-L2 antagonists include anti-PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that reduce, block, inhibit, eliminate or interfere with signal transduction derived from the interaction of PD-L2 with one or more of its binding partners (such as PD-1). In one embodiment, a PD-L2 binding antagonist reduces negative co-stimulatory signals mediated by or via cell surface proteins expressed on T lymphocytes (via PD-L2 mediated signaling), thereby making dysfunctional T cells less dysfunctional (e.g., enhancing effector responses to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0141] As used herein, the term "OX40" refers to any native OX40 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. The term encompasses "full-length," unprocessed OX40 as well as any form of OX40 resulting from processing in cells. The term also encompasses naturally occurring variants of OX40, such as splice variants or allelic variants. The amino acid sequence of an exemplary human OX40 is shown in SEQ ID NO: 354.

[0142] "OX40 activation" refers to activation of the OX40 receptor. Typically, OX40 activation results in signal transduction.

[0143] The term "aptamer" refers to a nucleic acid molecule that is capable of binding to a target molecule (such as a polypeptide). For example, the aptamer of the present invention can specifically bind to a TIGIT polypeptide, or a molecule in a signaling pathway that regulates TIGIT expression. The generation and therapeutic use of aptamers is well established in the art. See, for example, U.S. Patent No. 5,475,096, and (Eyetech, New York) for the treatment of age-related macular degeneration.

[0144] The term "dysfunction" in the context of immune dysfunction refers to a state of reduced immune responsiveness to antigenic stimulation.

[0145] As used herein, the term "dysfunctional" also includes insensitivity or non-responsiveness to antigen recognition, in particular, an impaired ability to translate antigen recognition into downstream T cell effector functions, such as proliferation, cytokine production (e.g., gamma interferon) and / or target cell killing.

[0146] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted immunoglobulins that bind to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages) enable these cytotoxic effector cells to specifically bind to antigen-bearing target cells and subsequently kill the target cells with cytotoxins. The primary cells mediating ADCC, NK cells, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991), page 464, Table 3 summarizes FcR expression on hematopoietic cells. To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. Nos. 5,500,362 or 5,821,337 or U.S. Pat. No. 6,737,056 (Presta), can be performed. Effector cells that can be used in such assays include PBMCs and NK cells. Alternatively, or in addition, ADCC activity of the target molecule can be assessed in vivo, for example, in an animal model such as that disclosed in Clynes et al., PNAS (USA) 95: 652-656 (1998). An exemplary assay for assessing ADCC activity is provided in the Examples herein.

[0147] The term "anergy" refers to incomplete or insufficient signaling (e.g., intracellular Ca 2+ in the absence of ras activation) that results from incomplete or insufficient signaling delivered via the T cell receptor. +2 T cell anergy can also occur after stimulation with an antigen in the absence of co-stimulation, generating cells that become insensitive to subsequent activation of the antigen even in the context of co-stimulation. The anergic state can often be overcome by the presence of interleukin-2 (IL-2). Anergic T cells do not undergo clonal expansion and / or acquire effector functions.

[0148] "Enhancing T cell function" means inducing, eliciting or stimulating effector or memory T cells to have renewed, sustained or amplified biological functions. Examples of enhancing T cell function include: increased levels of CD8 T cells relative to such levels before intervention. + Effector T cell secretion of interferon-γ, elevated from CD4 + Interferon-γ secretion by memory and / or effector T cells, elevated CD4 +Proliferation of effector and / or memory T cells, elevated CD8 + Proliferation of effector T cells, increased antigen responsiveness (e.g., clearance). In one embodiment, the level of enhancement is at least 50%, or 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%. Ways to measure this enhancement are known to those of ordinary skill in the art.

[0149] The term "exhaustion" refers to T cell exhaustion as a state of T cell dysfunction caused by continuous TCR signaling that occurs during many chronic infections and cancers. It is different from anergy in that it does not occur via incomplete or lacking signaling, but rather due to continuous signaling. It is defined by poor effector function, continuous inhibitory receptor expression, and a transcriptional state that is different from that of functional effector or memory T cells. Exhaustion prevents optimal control of infection and tumors. Exhaustion can be derived from both extrinsic negative regulatory pathways (such as immunomodulatory cytokines) and cell-intrinsic negative regulatory (co-stimulatory) pathways (PD-1, B7-H3, B7-H4, etc.).

[0150] "Enhancing T cell function" means inducing, causing or stimulating T cells to have sustained or amplified biological functions, or restoring or reactivating exhausted or inactive T cells. Examples of enhancing T cell function include: increased expression of CD8 T cells relative to such levels before intervention. + T cell secretion of gamma interferon, increased proliferation, increased antigen responsiveness (e.g., viral, pathogen, or tumor clearance). In one embodiment, the level of enhancement is at least 50%, or 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%. Methods for measuring this enhancement are known to those of ordinary skill in the art.

[0151] A "T cell dysfunction disorder" is a T cell disorder or condition characterized by reduced responsiveness to antigenic stimulation. In a specific embodiment, a T cell dysfunction disorder is a disorder specifically associated with inappropriately reduced signaling via OX40 and / or OX40L. In another embodiment, a T cell dysfunction disorder is a disorder in which T cells are anergic or have reduced ability to secrete cytokines, proliferate, or perform cytolytic activity. In a specific aspect, the reduced responsiveness results in ineffective control of pathogens or tumors expressing the immunogen. Examples of T cell dysfunction disorders characterized by T cell dysfunction include unresolved acute infections, chronic infections, and tumor immunity.

[0152] "Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Thus, as a therapeutic concept, tumor immunity is "treated" when this evasion is compromised, allowing the tumor to be recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

[0153] "Immunogenicity" refers to the ability of a particular substance to elicit an immune response. Tumors are immunogenic, and enhancing tumor immunogenicity helps clear tumor cells through an immune response. Examples of enhancing tumor immunogenicity include, but are not limited to, treatment with OX40 binding agonists (e.g., anti-OX40 agonist antibodies) and TIGIT inhibitors (e.g., anti-TIGIT blocking antibodies).

[0154] A "sustained response" refers to a persistent effect on reducing tumor growth after treatment has ceased. For example, the tumor size can remain the same or smaller than its size at the beginning of the administration period. In some embodiments, the sustained response has a duration that is at least the same as the duration of treatment, at least 1.5 times, 2.0 times, 2.5 times, or 3.0 times the duration of treatment.

[0155] "Carrier" as used herein includes pharmaceutically acceptable carriers, excipients or stabilizers that are non-toxic to the cells or mammals to which they are exposed at the dosages and concentrations employed. Typically, a physiologically acceptable carrier is a pH buffered aqueous solution. Examples of physiologically acceptable carriers include buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN TM , polyethylene glycol (PEG) and PLURONICS TM .

[0156] "Package insert" means instructions customarily included in commercial packages of drugs, that contain information about the indications, usage, dosage, administration, contraindications, other drugs associated with the packaged product and / or warnings concerning the use of such drug.

[0157] As used herein, "treatment" refers to a clinical intervention designed to alter the natural course of the individual or cell being treated during the clinical pathological process. The desired effects of treatment include a reduction in the rate of disease progression, an improvement or alleviation of the disease state, and a regression or improved prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with cancer are alleviated or eliminated, including but not limited to a reduction in proliferation (or destruction) of cancerous cells, a reduction in symptoms from the disease, an improvement in the quality of life of those individuals with the disease, a reduction in the dosage of other drugs needed to treat the disease, a delay in the progression of the disease, and / or a prolonged survival of the individual.

[0158] As used herein, "delaying the progression of a disease" means postponing, hindering, slowing down, delaying, stabilizing, and / or slowing the development of a disease (such as cancer). Depending on the disease history and / or the individual being treated, this delay can be of varying lengths of time. As will be apparent to one skilled in the art, a sufficient or significant delay can essentially encompass prevention, as the individual does not develop the disease. For example, the development of advanced stage cancers, such as metastases, can be delayed.

[0159] As used herein, the term "reducing or inhibiting cancer recurrence" means reducing or inhibiting tumor or cancer recurrence or tumor or cancer progression.

[0160] As used herein, "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Included within this definition are benign and malignant cancers, as well as dormant tumors or micrometastases. Examples of cancer include, but are not limited to, carcinoma, myeloma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include multiple myeloma (MM), squamous cell carcinoma, lung cancer (including small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung), peritoneal cancer, hepatocellular carcinoma, gastric or stomach cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, various types of head and neck cancer, and B-cell lymphoma (including low-grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, storage diseases (bulky disease) NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), and Meigs syndrome.

[0161] The term "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein.

[0162] As used herein, "metastasis" refers to the spread of cancer from its primary site to other locations in the body. Cancer cells can break away from the primary tumor, infiltrate into lymph and blood vessels, circulate via the bloodstream, and grow in distal lesions (metastasis) in normal tissues elsewhere in the body. Metastasis can be local or distal. Metastasis is a continuous process that depends on the tumor cells falling off from the primary tumor, spreading via the bloodstream, and stopping at the distal site. At the new site, the cells establish a blood supply and can grow to form a life-threatening mass. Excitatory and inhibitory molecular pathways in tumor cells regulate this behavior, and the interaction between tumor cells and host cells in the distal site is also important.

[0163] "Effective amount" is at least the minimum concentration required to achieve a measurable improvement or prevention of a particular condition. The effective amount herein can vary with factors such as the patient's disease state, age, sex, and weight, and the ability of the antibody to elicit a desired response in an individual. An effective amount is also the amount at which the therapeutic beneficial effect exceeds any toxic or adverse effect of the treatment. For preventive use, the beneficial or desired result includes the following result, such as eliminating or reducing risk, alleviating severity, or delaying the onset of the disease, including the biochemistry of the disease, histology and / or behavioral symptoms, the intermediate pathological phenotype presented during its complication and disease formation. For therapeutic use, the beneficial or desired result includes clinical results, such as reducing one or more symptoms from the disease, improving the quality of life of those subjects suffering from the disease, reducing the dosage of other drugs needed for treating the disease, enhancing the effect of another drug (such as via targeting), delaying the progression of the disease, and / or prolonging survival. In the case of cancer or tumors, an effective amount of a drug can have an effect in reducing the number of cancer cells; reducing tumor size; inhibiting (i.e., slowing down or desirably stopping) cancer cell infiltration into peripheral organs; inhibiting (i.e., slowing down and desirably stopping) tumor metastasis; inhibiting tumor growth to a certain extent; and / or alleviating one or more symptoms associated with the condition to a certain extent. An effective amount can be administered in one or more administrations. For the purposes of the present invention, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve a preventive or therapeutic treatment, directly or indirectly. As understood in a clinical context, an effective amount of a drug, compound, or pharmaceutical composition can be achieved with or without another drug, compound, or pharmaceutical composition. Thus, an "effective amount" can be considered in the context of administering one or more therapeutic agents, and a single agent can be considered to be administered in an effective amount if, together with one or more other agents, the desired result can be achieved or achieved.

[0164] As used herein, "in conjunction with" refers to administering a therapeutic modality in addition to another therapeutic modality. Thus, "in conjunction with" refers to administering another therapeutic modality before, during, or after administering one therapeutic modality to a subject.

[0165] As used herein, "subject" or "individual" refers to a mammal, including but not limited to a human or non-human mammal, such as a cow, horse, dog, sheep, or cat. Preferably, the subject refers to a human. In this article, a patient is also a subject.

[0166] "Chemotherapeutic agents" include chemical compounds useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib ( Genentech / OSI Pharm.), bortezomib ( Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant ( AstraZeneca, sunitinib ( Pfizer / Sugen, letrozole ( Novartis, imatinib mesylate ( Novartis), finasunate( Novartis, oxaliplatin ( Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (Sirolimus, Wyeth, Lapatinib ( GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336), sorafenib ( Bayer Labs, gefitinib ( AstraZeneca), AG1478, alkylating agents such as thiotepa and cyclosphosphamide; alkylsulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trihydroxymethylmelamine; rimethylomelamine; acetogenins (especially bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); adrenocorticosteroids, including prednisone and prednisolone; cyproterone acetate acetate); 5-alpha-reductase inhibitors, including finasteride and dutasteride; vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatin; aldesleukin, talc, duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin;Nitrogen mustards, such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard, mustard); nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma and calicheamicin omega (Angew Chem. Intl. Ed. Engl. 1994, 33: 183-186); anthracyclines (dynemicins), including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycins, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diaza-5-oxo-L-norleucine; (doxorubicin, morpholinodoxorubicin, cyanomorpholinodoxorubicin, 2-pyrrolidoxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteroyltriglutamic acid, purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, propionate, epitiostanol, mepitiostane, testolactone; antiadrenal drugs such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as frolinic acid; aceglatone;Aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamnol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as TAXOL (paclitaxel; Bristol-Myers Squibb Co., Ltd. Oncology, Princeton, NJ), (without Cremophor), an albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and Taxotere (docetaxel, doxetaxel; Sanofi-Aventis); chloranmbucil; (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine Ibandronate; CPT-11; the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; and pharmaceutically acceptable salts, acids, and derivatives of any of the foregoing.

[0167] Chemotherapeutic agents also include (i) antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and (toremifine citrate); (ii) aromatase inhibitors that inhibit the aromatase enzyme that regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazole, aminoglutethimide, (megestrol acetate), (exemestane; Pfizer), formestane, fadrozole, (vorozole), (letrozole; Novartis), and (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, and dapoxetine. acetate), diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signaling pathways involved in abnormal cell proliferation, such as, for example, PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (e.g., ) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, e.g. and rIL-2; topoisomerase 1 inhibitors, such as rmRH; and (ix) and pharmaceutically acceptable salts, acids and derivatives of any of the foregoing agents.

[0168] Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab ( Genentech; cetuximab ( Imclone; panitumumab ( Amgen, rituximab ( Genentech / Biogen Idec, pertuzumab ( 2C4, Genentech), trastuzumab ( Genentech), tositumomab (Bexxar, Corixia), and the antibody drug conjugate, gemtuzumab ozogamicin ( Wyeth). Other humanized monoclonal antibodies with therapeutic potential as agents for combination with the compounds of the invention include: apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, certolizumabpegol, cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab ), pascolizumab, pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab,Tucotuzumab celmoleukin, tucusituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), a recombinant proprietary human sequence full-length IgG1λ antibody genetically modified to recognize the interleukin-12p40 protein.

[0169] Chemotherapeutic agents also include "EGFR inhibitors," which refer to compounds that bind to EGFR or otherwise directly interact with EGFR and prevent or reduce its signaling activity, also referred to as "EGFR antagonists." Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimerized 225 (C225 or cetuximab; ) and remodeled human 225 (H225) (see WO96 / 40210, Imclone Systems Inc.); IMC-11F8, a fully human EGFR-targeting antibody (Imclone); antibodies that bind to type II mutant EGFR (U.S. Patent No. 5,212,290); humanized and chimeric antibodies that bind to EGFR, as described in U.S. Patent No. 5,891,996; and human antibodies that bind to EGFR, such as ABX-EGF or panitumumab (Panitumumab) (see WO98 / 50433, Abgenix / Amgen); EMD55900 (Stragliotto et al. al., Eur. J. Cancer 32A:636-640 (1996)); EMD7200 (matuzumab), a humanized EGFR antibody that is directed against EGFR and competes with both EGF and TGF-α for EGFR binding (EMD / Merck); a human EGFR antibody, HuMax-EGFR (GenMab); fully human antibodies designated E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3 and described in US 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated to cytotoxic agents to generate immunoconjugates (see, e.g., EP 659,439 A2, Merck Patent GmbH). EGFR antagonists include small molecules such as those described in U.S. Patent Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, 5,866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, and compounds described in PCT publications WO98 / 14451, WO98 / 50038, WO99 / 09016, and WO99 / 24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, Genentech / OSI Pharmaceuticals); PD 183805 (CI 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib 4-(3'-Chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib ( GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2-methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine).

[0170] Chemotherapeutic agents also include "tyrosine kinase inhibitors," including the EGFR-targeted drugs mentioned in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors, such as TAK165, available from Takeda; CP-724,714, an oral ErbB2 receptor tyrosine kinase selective inhibitor (Pfizer and OSI); dual HER inhibitors that preferentially bind to EGFR but inhibit both HER2 and EGFR-overexpressing cells, such as EKB-569 (available from Wyeth); lapatinib (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); Raf-1 inhibitors, such as rituximab (RI-1033; Pharmacia); and HER2 inhibitors. Pharmaceuticals, ISIS-5132, an antisense agent that inhibits Raf-1 signaling; non-HER-targeted TK inhibitors, such as imatinib mesylate ( available from Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors such as sunitinib ( available from Pfizer); VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular-regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines such as CGP 59326, CGP 60261, and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidines; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)-phthalimide); tyrphostine containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to HER-encoding nucleic acids); quinoxalines (U.S. Patent No. 5,804,396); tryphostins (U.S. Patent No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); imatinib mesylate PKI166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); INC-1C11 (Imclone), rapamycin (sirolimus, ); or any of the following patent publications: U.S. Patent No. 5,804,396; WO1999 / 09016 (American Cyanamid); WO1998 / 43960 (American Cyanamid); WO1997 / 38983 (Warner Lambert); WO1999 / 06378 (Warner Lambert); WO1999 / 06396 (Warner Lambert); WO1996 / 30347 (Pfizer, Inc); WO1996 / 33978 (Zeneca); WO1996 / 3397 (Zeneca) and WO1996 / 33980 (Zeneca).

[0171] Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, and dapoxetine. alfa, erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, interferon alfa-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium sodium), quinacrine, rasburicase, sargramostim, temozolomide, VM-26,6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate, and zoledronic acid, and pharmaceutically acceptable salts thereof.

[0172] Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, and dapoxetine. phosphate), fluocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolone caproate, fluocortolone pivalate, and fluprednidene acetate. acetate); immunoselective anti-inflammatory peptides (ImSAIDs), such as phenylalanine-glutamine-glycine (FEG) and its D-isomer form (feG) (IMULAN BioTherapeutics, LLC);antirheumatic drugs such as azathioprine, ciclosporin (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, sulfasalazine, tumor necrosis factor alpha (TNFα) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi), interleukin-1 (IL-1) blockers such as anakinra (Kineret), T-cell costimulation blockers such as abatacept (Orencia), interleukin-6 (IL-6) blockers such as tocilizumab; Interleukin-13 (IL-13) blockers, such as lebrikizumab; interferon alpha (IFN) blockers, such as Rontalizumab; β7-integrin blockers, such as rhuMAb Beta7; IgE pathway blockers, such as anti-M1 prime; secreted homotrimeric LTa3 and membrane-bound heterotrimeric LTa1 / β2 blockers, such as anti-lymphotoxin alpha (LTa); radioactive isotopes, such as At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 ,Bi 212 , P 32 , Pb 212and Lu radioisotopes; miscellaneous investigational agents such as thioplatin, PS-341, phenyl butyrate, ET-18-OCH3, or farnesyltransferase inhibitors (L-739749, L-744832; polyphenols such as quercetin, resveratrol, piceatannol, epigallocatechine gallate, theaflavin, flavanols, procyanidins, betulinic acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, ), β-lapachone, lapachol, colchicines, betulinic acid, acetylcamptothecin, scopolectin, and 9-aminocamptothecin, podophyllotoxin, and tegafur bexarotene Bisphosphonates, such as clodronate (e.g. or ), etidronate NE-58095, zoledronic acid / zoledronate alendronate Pamidronate tiludronate or risedronate and epidermal growth factor receptor (EGF-R); vaccines such as Vaccines; perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors such as oblimersen sodium Pixantrone; farnesyltransferase inhibitors, such as lonafarnib (SCH 6636, SARASAR TM ); and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing; and combinations of two or more of the foregoing, such as CHOP (an abbreviation for cyclophosphamide, doxorubicin, vincristine, and prednisolone combination therapy) and FOLFOX (oxaliplatin (ELOXATIN TM ) is the abbreviation for the combined treatment regimen of 5-FU and folinic acid).

[0173] Chemotherapeutic agents also include nonsteroidal anti-inflammatory drugs (NSAIDs) which have analgesic, antipyretic and anti-inflammatory effects. NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include aspirin, propionic acid derivatives such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin and naproxen, acetic acid derivatives such as indomethacin, sulindac, etodolac, diclofenac, enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam and isoxicam, fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid and fenamic acid. NSAIDs are indicated for the relief of symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, pain from bone metastases, headaches and migraines, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction, and renal colic.

[0174] As used herein, the term "cytokine" generally refers to a protein released by a cell population that acts as an intercellular mediator on another cell or has an autocrine effect on the cell that produces the protein. Examples of such cytokines include lymphokines, monokines; interleukins ("IL") such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL10, IL-11, IL-12, IL-13, IL-15, IL-17A-F, IL-18 to IL-29 (such as IL-23), IL-31, including rIL-2; tumor necrosis factor, such as TNF-α or TNF-β, TGF-β1-3; and other polypeptide factors, including leukemia inhibitory factor ("LIF"), ciliary ganglion neurotrophic factor ("CNTF"), CNTF-like cytokine ("CLC"), cardiotrophin ("CT"), and kit ligand ("KL").

[0175] As used herein, the term "chemokine" refers to a soluble factor (e.g., cytokine) that has the ability to selectively induce chemotaxis and activation of leukocytes. They also trigger processes such as angiogenesis, inflammation, wound healing, and tumorigenesis. Examples of chemokines include IL-8, the human homolog of mouse keratinocyte chemoattractant (KC).

[0176] "Percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to obtain maximum percent sequence identity, and without considering any conservative substitutions as part of the sequence identity. Comparisons for the purpose of determining percent amino acid sequence identity can be performed in a variety of ways within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required to achieve maximum alignment over the full length of the compared sequences. However, for the purposes of the present invention, % amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc., and the source code has been submitted, along with user documentation, to the U.S. Copyright Office (US Copyright Office, Washington DC, 20559), where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from source code. The ALIGN-2 program should be compiled for use on UNIX operating systems, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and are unchanged.

[0177] In the case of using ALIGN-2 to compare amino acid sequences, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (or can be expressed as a given amino acid sequence A having or comprising a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows:

[0178] Multiply the fraction X / Y by 100

[0179] where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, then the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A. Unless expressly stated otherwise, all % amino acid sequence identity values ​​used herein are obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[0180] The phrase "pharmaceutically acceptable" indicates that the substance or composition must be compatible chemically and / or toxicologically with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0181] As used herein, "administering" refers to a method of administering a dose of a compound (e.g., an anti-TIGIT antibody of the invention or a nucleic acid encoding an anti-TIGIT antibody of the invention) or a composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition comprising an anti-TIGIT antibody of the invention) to a subject. The compositions utilized in the methods described herein can be administered, for example, intravenously, intramuscularly, intravitreally (e.g., by intravitreal injection), by eye drops, intradermally, subcutaneously, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intrathecally, intranasally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intracapsularly, intramucosally, intrapericardially, intraumbilically, intraocularly, intraorbitally, orally, topically, transdermally, by inhalation, by injection, by implantation, by infusion, by continuous infusion, by local perfusion of target cells by direct immersion, by catheter, by lavage, as a cream, or as a lipid composition. The compositions utilized in the methods described herein can also be administered systemically or topically. The method of administration may vary depending on factors such as the compound or composition being administered and the severity of the disorder, disease, or condition being treated.

[0182] As used herein, the term "vector" means a nucleic acid molecule capable of transporting other nucleic acids connected thereto. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop to which another DNA segment can be connected. Another type of vector is a bacteriophage vector. Another type of vector is a viral vector, in which another DNA segment can be connected to a viral genome. Some vectors can replicate autonomously in the host cell they are introduced into (e.g., bacterial vectors and additional mammalian vectors with bacterial replication origins). Other vectors (e.g., non-additional mammalian vectors) can be integrated into the genome of the host cell after being introduced into the host cell, thereby replicating together with the host genome. In addition, some vectors can guide the expression of genes operably connected thereto. Such vectors are referred to herein as "recombinant expression vectors" (or simply "recombinant vectors" or "expression vectors"). Generally, expression vectors useful in recombinant DNA technology are often in the form of plasmids. In this manual, "plasmid" and "vector" are used interchangeably.

[0183] As used herein, the term "about" refers to the normal error range of the corresponding numerical value that is readily known to those skilled in the art. Reference herein to "about" a numerical value or parameter includes (and describes) embodiments involving the numerical value or parameter itself.

[0184] III. Exemplary anti-TIGIT antibodies

[0185] The present invention provides anti-TIGIT antibodies useful for treating or delaying progression of cancer or an immune-related disease (eg, a T-cell dysfunctional disorder) in a subject (eg, a human).

[0186] In one example, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more amino acid residues (e.g., 1, 2, or 3 amino acid residues) selected from the group consisting of Ser78, Ser80, and Lys82 of human TIGIT. For example, in some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Ser80 and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Ser78 and Ser80. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Ser78 and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Ser78, Ser80, and Lys82.

[0187] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes residue Ala67 of human TIGIT in addition to Ser78, Ser80, and / or Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78 and Ala67; Ser80 and Ala67; Lys82 and Ala67; Ser78, Ser80, and Ala67; Ser78, Lys82, and Ala67; Ser80, Lys82, and Ala67; or Ser78, Ser80, Lys82, and Ala67.

[0188] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, or 3 amino acid residues) other than Ser78, Ser80, and / or Lys82 selected from the group consisting of Glu60, Leu65, and Ile68 of human TIGIT. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78 and Glu60; Ser80 and Glu60; Lys82 and Glu60; Ser78 and Leu65; Ser80 and Leu65; Lys82 and Leu65; Ser78 and Ile68; Ser80 and Ile68; Lys82 and Ile68. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, and Glu60; Ser78, Ser80, and Leu65; Ser78, Ser80, and Ile68; Ser80, Lys82, and Glu60; Ser80, Lys82, and Leu65; Ser80, Lys82, and Ile68; Ser78, Lys82, and Glu60; Ser78, Lys82, and Leu65; Ser78, Lys82, and Ile68. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, and Glu60; Ser78, Ser80, Lys82, and Leu65; Ser78, Ser80, Lys82, and Ile68. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Glu60, Leu65, and Ser78; Glu60, Leu65, and Ser80; Glu60, Leu65, and Lys82; Leu65, Ile68, and Ser78; Leu65, Ile68, and Ser80; Leu65, Ile68, and Lys82; Glu60, Ile68, and Ser78; Glu60, Ile68, and Ser80; Glu60, Ile68, and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Glu60, Leu65, Ile68, and Ser78; Glu60, Leu65, Ile68, and Ser80; Glu60, Leu65, Ile68, and Lys82.In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Glu60, Leu65, Ser78, and Ser80; Glu60, Leu65, Ser78, and Lys82; Glu60, Leu65, Ser80, and Lys82; Leu65, Ile68, Ser78, and Ser80; Leu65, Ile68, Ser78, and Lys82; Leu65, Ile68, Ser80, and Lys82; Leu65, Glu60, Ser78, and Ser80; Leu65, Glu60, Ser78, and Lys82; Leu65, Glu60, Ser80, and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Glu60, Leu65, Ser78, Ser80, and Lys82; Leu65, Ile68, Ser78, Ser80, and Lys82; Glu60, Leu65, Ser78, Ser80, and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Glu60, Leu65, and Ile68; Ser78, Lys82, Glu60, Leu65, and Ile68; Ser80, and Lys82, Glu60, Leu65, and Ile68. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, Glu60, Leu65, and Ile68.

[0189] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, 3, or 4 amino acid residues) other than Ser78, Ser80, and Lys82 selected from the group consisting of Gln56, Asn70, Leu73, and His111 of human TIGIT. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, and Gln56; Ser78, Ser80, Lys82, and Asn70; Ser78, Ser80, Lys82, and Leu73; Ser78, Ser80, Lys82, and His111. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, Gln56, and Asn70; Ser78, Ser80, Lys82, Gln56, and Leu73; Ser78, Ser80, Lys82, Gln56, and His111; Ser78, Ser80, Lys82, Asn70, and Leu73; Ser78, Ser80, Lys82, Asn70, and His111; Ser78, Ser80, Lys82, Leu73, and His111. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, Gln56, Asn70, and Leu73; Ser78, Ser80, Lys82, Gln56, Asn70, and His111; Ser78, Ser80, Lys82, Gln56, Leu73, and His111. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Ser78, Ser80, Lys82, Gln56, Asn70, Leu73, and His111. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, or 3 amino acid residues) other than Ser78, Ser80, and / or Lys82 selected from the group consisting of Thr55, Asn58, Asp63, Gln64, His76, Ile77, and Pro79 of human TIGIT.

[0190] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Thr55, Gln56, Asn58, Glu60, Asp63, Gln64, Leu65, Ala67, Ile68, Asn70, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that consists of amino acid residues Thr55, Gln56, Asn58, Glu60, Asp63, Gln64, Leu65, Ala67, Ile68, Asn70, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT.

[0191] In another example, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more amino acid residues (e.g., 1, 2, or 3 amino acid residues) selected from the group consisting of Thr55, Ser80, and Lys82 of human TIGIT. For example, in some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residue Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Ser80 and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Thr55 and Ser80. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Thr55 and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Thr55, Ser80, and Lys82.

[0192] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes residue Gln56 of human TIGIT in addition to Thr55, Ser80, and / or Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55 and Gln56; Ser80 and Gln56; Lys82 and Gln56; Thr55, Ser80, and Gln56; Thr55, Lys82, and Gln56; Ser80, Lys82, and Gln56; or Thr55, Ser80, Lys82, and Gln56.

[0193] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, or 3 amino acid residues) other than Thr55, Ser80, and / or Lys82 selected from the group consisting of Asn58, Glu60, Ile77, and Pro79 of human TIGIT. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55 and Asn58; Thr55 and Glu60; Ser80 and Asn58; Ser80 and Glu60; Lys82 and Asn58; Lys82 and Glu60; Thr55 and Ile77; Ser80 and Ile77; Lys82 and Ile77; Thr55 and Pro79; Ser80 and Pro79; Lys82 and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Ser80, and Asn58; Thr55, Ser80, and Glu60; Thr55, Ser80, and Ile77; Thr55, Ser80, and Pro79; Ser80, Lys82, and Asn58; Ser80, Lys82, and Glu60; Ser80, Lys82, and Ile77; Ser80, Lys82, and Pro79; Thr55, Lys82, and Asn58; Thr55, Lys82, and Glu60; Thr55, Lys82, and Ile77; Thr55, Lys82, and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Ser80, Lys82, and Asn58; Thr55, Ser80, Lys82, and Glu60; Thr55, Ser80, Lys82, and Ile77; Thr55, Ser80, Lys82, and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Asn58, Ile77, and Thr55; Glu60, Ile77, and Thr55; Asn58, Ile77, and Ser80; Glu60, Ile77, and Ser80; Asn58, Ile77, and Lys82; Glu60, Ile77, and Lys82; Ile77, Pro79, and Thr55; Ile77, Pro79, and Ser80; Ile77, Pro79, and Lys82; Glu60, Pro79, and Thr55; Asn58, Pro79, and Ser80; Glu60, Pro79, and Ser80; Asn58, Pro79, and Lys82; Glu60, Pro79, and Lys82.In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Asn58, Ile77, Pro79, and Thr55; Glu60, Ile77, Pro79, and Thr55; Asn58, Ile77, Pro79, and Ser80; Glu60, Ile77, Pro79, and Ser80; Asn58, Ile77, Pro79, and Lys82; Glu60, Ile77, Pro79, and Lys82. In some cases, the anti-TIGIT antibody binds to a region of human TIGIT comprising Asn58, Ile77, Thr55, and Ser80; Glu60, Ile77, Thr55, and Ser80; Asn58, Ile77, Thr55, and Lys82; Glu60, Ile77, Thr55, and Lys82; Asn58, Ile77, Ser80, and Lys82; Glu60, Ile77, Ser80, and Lys82; Ile77, Pro79, Thr55, and Ser80; Ile77, Pro79, T Epitopes of hr55, and Lys82; Ile77, Pro79, Ser80, and Lys82; Ile77, Asn58, Thr55, and Ser80; Ile77, Glu60, Thr55, and Ser80; Ile77, Asn58, Thr55, and Lys82; Ile77, Glu60, Thr55, and Lys82; Ile77, Asn58, Ser80, and Lys82; Ile77, Glu60, Ser80, and Lys82. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Asn58, Ile77, Thr55, Ser80, and Lys82; Glu60, Ile77, Thr55, Ser80, and Lys82; Ile77, Pro79, Thr55, Ser80, and Lys82; Asn58, Ile77, Thr55, Ser80, and Lys82; Glu60, Ile77, Thr55, Ser80, and Lys82.In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Ser80, Asn58, Ile77, and Pro79; Thr55, Ser80, Glu60, Ile77, and Pro79; Thr55, Lys82, Asn58, Ile77, and Pro79; Thr55, Lys82, Glu60, Ile77, and Pro79; Ser80, Lys82, Asn58, Ile77, and Pro79; and Ser80, Lys82, Glu60, Ile77, and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Ser80, Lys82, Glu60, Ile77, and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Ser80, Lys82, Asn58, Ile77, and Pro79. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Thr55, Asn58, Glu60, Ile77, Pro79, Ser80, and Lys82.

[0194] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, 3, 4, 5, or 6 amino acid residues) in addition to Thr55, Ser80, and Lys82 selected from the group consisting of Leu65, Ile68, Leu73, His76, Ser78, and His111 that further comprises amino acid residues Leu65, Ile68, Leu73, His76, Ser78, and His111 of human TIGIT.

[0195] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Thr55, Gln56, Asn58, Glu60, Leu65, Ile68, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that consists of amino acid residues Thr55, Gln56, Asn58, Glu60, Leu65, Ile68, Leu73, His76, Ile77, Ser78, Pro79, Ser80, Lys82, and His111 of human TIGIT.

[0196] In some cases, any of the anti-TIGIT antibodies described above may include a paratope comprising one or more amino acid residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acid residues) selected from the group consisting of heavy chain variable region amino acid residues Asn32, Tyr52, Arg52b, Phe53, Lys54, Tyr56, Asp58, Tyr99, AsplOO, LeulOOa, LeulOOb, and Ala100c and / or one or more amino acid residues (e.g., 1, 2, 3, 4, or 5 amino acid residues) selected from the group consisting of light chain variable region amino acid residues Tyr27d, Tyr92, Ser93, Thr94, and Phe96.

[0197] In some cases, the anti-TIGIT antibody may include a paratope consisting of heavy chain variable region amino acid residues Asn32, Tyr52, Arg52b, Phe53, Lys54, Tyr56, Asp58, Tyr99, Asp100, Leu100a, Leu100b, and Ala100c and light chain variable region amino acid residues Tyr27d, Tyr92, Ser93, Thr94, and Phe96.

[0198] In certain embodiments, an anti-TIGIT antibody of the present invention makes unique contacts with amino acids of human TIGIT at a distance of 4.5 angstroms, 3.7 angstroms, 3.5 angstroms, 3.25 angstroms, 3.00 angstroms, 2.75 angstroms, or less. In certain embodiments, an antibody is provided that binds to an epitope consisting of one, two, three, four, or five amino acids of human TIGIT at a distance of 4.5 angstroms, 3.7 angstroms, 3.5 angstroms, 3.25 angstroms, 3.00 angstroms, 2.75 angstroms or less. In one embodiment, an anti-TIGIT antibody of the present invention makes unique contacts with amino acids of human TIGIT at a distance of 3.7 angstroms or less. In certain embodiments, an antibody is provided that binds to an epitope consisting of one, two, three, four, or five amino acids of human TIGIT at a distance of 3.7 angstroms or less.

[0199] In some cases, any of the above anti-TIGIT antibodies includes at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of SNSAAWN (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 5); and / or (f) HVR-L3 comprising the amino acid sequence of QQYYSTPFT (SEQ ID NO: 6), or one or more of the above HVRs and their combinations with SEQ ID NO: 7. A combination of one or more variants of any of ID NOs: 1-6 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above-mentioned anti-TIGIT antibodies include (a) HVR-H1 comprising the amino acid sequence of SNSAAWN (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQYYSTPFT (SEQ ID NO: 6), such as those possessed by the anti-TIGIT antibody 4.1D3 and its derivatives (e.g., 4.1D3.Q1E). In some cases, the anti-TIGIT antibody may have a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 34 or 35, or the sequence of SEQ ID NO: 34 or 35 and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 36, or the sequence of SEQ ID NO: 36.In some cases, the anti-TIGIT antibody can have a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 34, or the sequence of SEQ ID NO: 34 and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 36, or the sequence of SEQ ID NO: 36. In a specific instance, the anti-TIGIT antibody can be 4.1D3.Q1E, or a derivative or clonal relative thereof. In some cases, the anti-TIGIT antibody can have a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 35, or the sequence of SEQ ID NO: 35 and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 36, or the sequence of SEQ ID NO: 36. In a specific instance, the anti-TIGIT antibody can be 4.1D3, or a derivative or clonal relative thereof.

[0200] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7); FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8); FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and / or FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 7-10. In some cases, for example, the antibody further comprises a FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7); a FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8); a FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and a FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10), such as those possessed by anti-TIGIT antibodies 4.1D3.Q1E and 4.1D3.

[0201] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11), wherein X1 is Q or E; FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 11-14. The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 15); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 12-15. In some cases, the anti-TIGIT antibody includes a FR-H1 comprising the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 15); a FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); a FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and a FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), such as that possessed by the 4.1D3.Q1E antibody.In another instance, for example, the anti-TIGIT antibody may further comprise at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 16); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 12-14 and 16. In some cases, the anti-TIGIT antibody includes a FR-H1 comprising the amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 16); a FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); a FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and a FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), such as that possessed by the 4.1D3 antibody.

[0202] In another example, the present invention provides an anti-TIGIT antibody that binds to an epitope on human TIGIT that includes one or more amino acid residues (e.g., 1, 2, or 3 amino acid residues) selected from the group consisting of Gln53, His111, and Tyr113 of human TIGIT. For example, in some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues His111 and Tyr113. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Gln53 and His111. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Gln53 and Tyr113. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes amino acid residues Gln53, His111, and Tyr113.

[0203] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes residue Gln56 of human TIGIT in addition to Gln53, His111, and / or Tyr113. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53 and Gln56; His111 and Gln56; Tyr113 and Gln56; Gln53, His111, and Gln56; Gln53, Tyr113, and Gln56; His111, Tyr113, and Gln56; or Gln53, His111, Tyr113, and Gln56.

[0204] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes one or more additional amino acid residues (e.g., 1, 2, 3, 4, 5, or 6 amino acid residues) other than Gln53, His111, Tyr113, and / or Gln56 selected from the group consisting of Glu60, Leu65, Ile68, Asn70, Leu73, and His76 of human TIGIT. For example, in some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, and Glu60; Gln53, His111, Tyr113, Gln56, and Leu65; Gln53, His111, Tyr113, Gln56, and Ile68; Gln53, His111, Tyr113, Gln56, and Asn70; Gln53, His111, Tyr113, Gln56, and Leu73; Gln53, His111, Tyr113, Gln56, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, and Leu65; Gln53, His111, Tyr113, Gln56, Glu60, and Ile68; Gln53, His111, Tyr113, Gln56, Glu60, and Asn70; Gln53, His111, Tyr113, Gln56, Glu60, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Leu65, and Ile68; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, and Asn70; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Ile68, and Asn70; Gln53, His111, Tyr113, Gln56, Glu60, Ile68, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Ile68, and His76.In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Asn70, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Leu73, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT including Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, and Asn70; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Ile68, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Ile68, Asn70, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT including Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, Asn70, and His76; Gln53, His111, Tyr113, Gln56, Glu60, Ile68, Asn70, Leu73 and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Glu60, Leu65, Ile68, Asn70, Leu73, and His76.

[0205] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, and Ile68; Gln53, His111, Tyr113, Gln56, Leu65, and Asn70; Gln53, His111, Tyr113, Gln56, Leu65, and Leu73; Gln53, His111, Tyr113, Gln56, Leu65, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Ile68, and Asn70; Gln53, His111, Tyr113, Gln56, Leu65, Ile68, and Leu73; Gln53, His111, Tyr113, Gln56, Leu65, Ile68, and Asn70; Gln53, His111, Tyr113, Gln56, Leu65, Ile68, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Leu65, Asn70, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Leu73, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Ile68, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Leu65, Ile68, Asn70, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Asn70, Leu73, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu65, Ile68, Asn70, Leu73, and His76.

[0206] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Ile68, and Asn70; Gln53, His111, Tyr113, Gln56, Ile68, and Leu73; Gln53, His111, Tyr113, Gln56, Ile68, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Ile68, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Ile68, Asn70, and His76; Gln53, His111, Tyr113, Gln56, Ile68, Leu73, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Ile68, Asn70, Leu73, and His76.

[0207] In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Asn70, and Leu73; Gln53, His111, Tyr113, Gln56, Asn70, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Asn70, Leu73, and His76. In some cases, the anti-TIGIT antibody binds to an epitope on human TIGIT that includes Gln53, His111, Tyr113, Gln56, Leu73, and His76.

[0208] In some cases, any of the anti-TIGIT antibodies of the foregoing embodiments comprises at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); (b) HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); (c) HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); (d) HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); (e) HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and / or (f) HVR-L3 comprising the amino acid sequence of QHLHGYPX1N (SEQ ID NO: 22), wherein X1 is C or S, or one or more of the above HVRs and their combinations with SEQ ID NO: A combination of one or more variants of any of NOs: 17-22 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above-mentioned anti-TIGIT antibodies of the foregoing embodiments may include, for example, (a) an HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); (c) an HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); (d) an HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); (e) an HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and (f) an HVR-L3 comprising the amino acid sequence of QHLHGYPX1N (SEQ ID NO: 22), such as those possessed by the anti-TIGIT antibody 7.4A3 and its derivatives (e.g., 7.4A3.C96S.Q1E).In some cases, the anti-TIGIT antibody may include (a) an HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); (c) an HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); (d) an HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); (e) an HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and (f) an HVR-L3 comprising the amino acid sequence of QHLHGYPSN (SEQ ID NO: 23), such as that possessed by the anti-TIGIT antibody 7.4A3.C96S.Q1E. In other cases, the anti-TIGIT antibody may include (a) an HVR-H1 comprising the amino acid sequence of SYPMN (SEQ ID NO: 17); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGNPTYVQGFTG (SEQ ID NO: 18); (c) an HVR-H3 comprising the amino acid sequence of TGGHTYDSYAFDV (SEQ ID NO: 19); (d) an HVR-L1 comprising the amino acid sequence of RASQVISSSLA (SEQ ID NO: 20); (e) an HVR-L2 comprising the amino acid sequence of AASTLQS (SEQ ID NO: 21); and (f) an HVR-L3 comprising the amino acid sequence of QHLHGYPCN (SEQ ID NO: 28), such as that possessed by the anti-TIGIT antibody 7.4A3.

[0209] In some cases, the anti-TIGIT antibody can have a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 37, or the sequence of SEQ ID NO: 37 and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 38, or the sequence of SEQ ID NO: 38. In a specific instance, the anti-TIGIT antibody can be 7.4A3.C96S.Q1E, or a derivative or clonal relative thereof. In some cases, the anti-TIGIT antibody can have a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 39, or the sequence of SEQ ID NO: 39 and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 40, or the sequence of SEQ ID NO: 40. In a specific instance, the anti-TIGIT antibody can be 7.4A3, or a derivative or clonal relative thereof.

[0210] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): FR-L1 comprising the amino acid sequence of DIQLTQSPTFLSASVGDRVTITC (SEQ ID NO: 30); FR-L2 comprising the amino acid sequence of WYQQNPGKAPKLLIY (SEQ ID NO: 31); FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEFTLTISSLQPEDFVTYYC (SEQ ID NO: 32); and / or FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 33), or one or more of the above FRs and their combinations with SEQ ID NO: A combination of one or more variants of any of NO:30-33 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by the anti-TIGIT antibody 7.4A3 and its derivatives (e.g., 7.4A3.C96S.Q1E).

[0211] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 24) or the amino acid sequence of QVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 29); FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and / or FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27), or one or more of the above FRs and their combinations with SEQ ID NO: 28. A combination of one or more variants of any of NOs: 24-27 and 29 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 24); FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and / or FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 24-27. In some cases, the anti-TIGIT antibody includes a FR-H1 comprising the amino acid sequence of EVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 24); a FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); a FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and a FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27), such as that possessed by the 7.4A3.C96S.Q1E antibody.In another instance, for example, the anti-TIGIT antibody may further include at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 29); FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and / or FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 25-27 and 29. In some cases, the anti-TIGIT antibody includes a FR-H1 comprising the amino acid sequence of QVQLVQSGSDLKKPGASVRVSCKASGYTFT (SEQ ID NO: 29); a FR-H2 comprising the amino acid sequence of WVRQAPGHGLEWMG (SEQ ID NO: 25); a FR-H3 comprising the amino acid sequence of RFVFSLDTSVNTAYLQISSLKAEDTAVYFCAR (SEQ ID NO: 26); and a FR-H4 comprising the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 27), such as possessed by the 7.4A3 anti-TIGIT antibody.

[0212] In another example, the present invention provides an anti-TIGIT antibody comprising at least one, two, three, four, five, or six HVRs selected from the following: (a) an HVR-H1 comprising the amino acid sequence of NYPMN (SEQ ID NO: 41); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGSPAYAQDFTE (SEQ ID NO: 42); (c) an HVR-H3 comprising the amino acid sequence of TAITSVYHFDY (SEQ ID NO: 43); (d) an HVR-L1 comprising the amino acid sequence of RASQGISSYLA (SEQ ID NO: 44); (e) an HVR-L2 comprising the amino acid sequence of GATTLQS (SEQ ID NO: 45); and / or (f) an HVR-L3 comprising the amino acid sequence of QKLNSHPX1S (SEQ ID NO: 46), wherein X1 is C, S, or Y, or one or more of the above HVRs and their combinations with SEQ ID NO: NO: 41-46 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above-mentioned anti-TIGIT antibodies of the aforementioned embodiments may include, for example, (a) HVR-H1 comprising the amino acid sequence of NYPMN (SEQ ID NO: 41); (b) HVR-H2 comprising the amino acid sequence of WINTNTGSPAYAQDFTE (SEQ ID NO: 42); (c) HVR-H3 comprising the amino acid sequence of TAITSVYHFDY (SEQ ID NO: 43); (d) HVR-L1 comprising the amino acid sequence of RASQGISSYLA (SEQ ID NO: 44); (e) HVR-L2 comprising the amino acid sequence of GATTLQS (SEQ ID NO: 45); and (f) HVR-H3 comprising the amino acid sequence of QKLNSHPX1S (SEQ ID NO: 46). NO:46), wherein X1 is C, S, or Y, such as possessed by the anti-TIGIT antibody 4.1A4 and its derivatives (e.g., 4.1A4.C96S.Q1E and 4.1A4.C96Y.Q1E).In some cases, the anti-TIGIT antibody may include (a) an HVR-H1 comprising the amino acid sequence of NYPMN (SEQ ID NO: 41); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGSPAYAQDFTE (SEQ ID NO: 42); (c) an HVR-H3 comprising the amino acid sequence of TAITSVYHFDY (SEQ ID NO: 43); (d) an HVR-L1 comprising the amino acid sequence of RASQGISSYLA (SEQ ID NO: 44); (e) an HVR-L2 comprising the amino acid sequence of GATTLQS (SEQ ID NO: 45); and (f) an HVR-L3 comprising the amino acid sequence of QKLNSHPCS (SEQ ID NO: 47), such as that possessed by the anti-TIGIT antibody 4.1A4. In other cases, the anti-TIGIT antibody may include (a) an HVR-H1 comprising the amino acid sequence of NYPMN (SEQ ID NO: 41); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGSPAYAQDFTE (SEQ ID NO: 42); (c) an HVR-H3 comprising the amino acid sequence of TAITSVYHFDY (SEQ ID NO: 43); (d) an HVR-L1 comprising the amino acid sequence of RASQGISSYLA (SEQ ID NO: 44); (e) an HVR-L2 comprising the amino acid sequence of GATTLQS (SEQ ID NO: 45); and (f) an HVR-L3 comprising the amino acid sequence of QKLNSHPSS (SEQ ID NO: 48), such as that possessed by the anti-TIGIT antibody 4.1A4.C96S.Q1E. In other cases, the anti-TIGIT antibody may include (a) an HVR-H1 comprising the amino acid sequence of NYPMN (SEQ ID NO: 41); (b) an HVR-H2 comprising the amino acid sequence of WINTNTGSPAYAQDFTE (SEQ ID NO: 42); (c) an HVR-H3 comprising the amino acid sequence of TAITSVYHFDY (SEQ ID NO: 43); (d) an HVR-L1 comprising the amino acid sequence of RASQGISSYLA (SEQ ID NO: 44); (e) an HVR-L2 comprising the amino acid sequence of GATTLQS (SEQ ID NO: 45); and (f) an HVR-L3 comprising the amino acid sequence of QKLNSHPYS (SEQ ID NO: 49), such as that possessed by the anti-TIGIT antibody 4.1A4.C96Y.Q1E.

[0213] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 56); FR-L2 comprising the amino acid sequence of WYQQKPGKAPRVLIY (SEQ ID NO: 57); FR-L3 comprising the amino acid sequence of GVPSRFSGSESGTEFTLTISSLQPEDLATYYC (SEQ ID NO: 58); and / or FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 59), or one or more of the above FRs and their combinations with SEQ ID NO: 50. A combination of one or more variants of any of NO:56-59 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by the anti-TIGIT antibody 4.1A4 and its derivatives (e.g., 1A4.C96S.Q1E and 1A4.C96Y.Q1E).

[0214] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of X1VQLVQSGSELKKPGASVKVSCKASGYTLT (SEQ ID NO: 50), wherein X1 is E or Q; FR-H2 comprising the amino acid sequence of WVRQAPGRGLEWMG (SEQ ID NO: 51); FR-H3 comprising the amino acid sequence of RFVFSLDTSVTTAYLQISSLKAEDTAVYYCAR (SEQ ID NO: 52); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 53), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 50-53. The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVQLVQSGSELKKPGASVKVSCKASGYTLT (SEQ ID NO: 54); FR-H2 comprising the amino acid sequence of WVRQAPGRGLEWMG (SEQ ID NO: 51); FR-H3 comprising the amino acid sequence of RFVFSLDTSVTTAYLQISSLKAEDTAVYYCAR (SEQ ID NO: 52); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 53), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 51-54, such as those possessed by the anti-TIGIT antibody 4.1A4.In some cases, the anti-TIGIT antibody comprises an FR-H1 comprising the amino acid sequence of EVQLVQSGSELKKPGASVKVSCKASGYTLT (SEQ ID NO: 55); an FR-H2 comprising the amino acid sequence of WVRQAPGRGLEWMG (SEQ ID NO: 51); an FR-H3 comprising the amino acid sequence of RFVFSLDTSVTTAYLQISSLKAEDTAVYYCAR (SEQ ID NO: 52); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 53), or one or more of the above FRs and their combinations with SEQ ID NO: 54. A combination of one or more variants of any of IDNOs: 51-53 and 55 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as possessed by anti-TIGIT antibody 1A4.C96S.Q1E or 1A4.C96Y.Q1E.

[0215] In another example, the present invention provides an anti-TIGIT antibody comprising at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of TYGMGVS (SEQ ID NO: 68); (b) HVR-H2 comprising the amino acid sequence of SIWWNGNTYYNPSLKS (SEQ ID NO: 69); (c) HVR-H3 comprising the amino acid sequence of TGGAVITWFAY (SEQ ID NO: 70); (d) HVR-L1 comprising the amino acid sequence of KASQSVGKNIA (SEQ ID NO: 71); (e) HVR-L2 comprising the amino acid sequence of YASNRYT (SEQ ID NO: 72); and / or (f) HVR-L3 comprising the amino acid sequence of QHIYNSPYP (SEQ ID NO: 73), or one or more of the above HVRs and their combinations with SEQ ID NO: NO: 68-73 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above-mentioned anti-TIGIT antibodies of the aforementioned embodiments may include, for example, (a) HVR-H1 comprising the amino acid sequence of TYGMGVS (SEQ ID NO: 68); (b) HVR-H2 comprising the amino acid sequence of SIWWNGNTYYNPSLKS (SEQ ID NO: 69); (c) HVR-H3 comprising the amino acid sequence of TGGAVITWFAY (SEQ ID NO: 70); (d) HVR-L1 comprising the amino acid sequence of KASQSVGKNIA (SEQ ID NO: 71); (e) HVR-L2 comprising the amino acid sequence of YASNRYT (SEQ ID NO: 72); and (f) HVR-H3 comprising the amino acid sequence of QHIYNSPYP (SEQ ID NO: 73). NO:73), such as that possessed by the anti-TIGIT antibody rat6B2 and its derivatives (e.g., h6B2.L1H1, h6B2.L2H1, h6B2.L1H2, h6B2.L1H3, h6B2.L1H4, h6B2.L1H5, and h6B2.L2H5).

[0216] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): an FR-L1 comprising the amino acid sequence of XIIX2MTQSPX3SX4SX5SVGDRVTX6X7C (SEQ ID NO: 88), wherein X1 is D or N; X2 is Q or V; X3 is K or S; X4 is L or M; X5 is A or I; X6 is I or M; and X7 is N or T; an FR-L2 comprising the amino acid sequence of WYQQKPGX1X2PKLLIY (SEQ ID NO: 89), wherein X1 is K or Q and X2 is A or S; and an FR-L3 comprising the amino acid sequence of GVPX1RFX2GX3GSGTDFTX4TIX5X6X7QX8EDX9AX 10 X 11 FR-L3 of the amino acid sequence of YC (SEQ ID NO: 90), wherein X1 is D or S; X2 is S or T; X3 is G or S; X4 is F or L; X5 is N or S; X6 is S or T; X7 is L or V; X8 is A or P; X9 is A or I; X 10 is F or T; and X 11is F or Y; and / or a FR-L4 comprising the amino acid sequence of FGX1GTKX2EIK (SEQ ID NO: 91), wherein X1 is Q or T and X2 is L or V, or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 88-91, such as those possessed by the anti-TIGIT antibody rat6B2 and derivatives thereof (e.g., h6B2.L1H1, h6B2.L2H1, h6B2.L1H2, h6B2.L1H3, h6B2.L1H4, h6B2.L1H5, and h6B2.L2H5). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following light chain variable region FRs: FR-L1 comprising the amino acid sequence of NIVMTQSPKSMSISVGDRVTMNC (SEQ ID NO: 92); FR-L2 comprising the amino acid sequence of WYQQKPGQSPKLLIY (SEQ ID NO: 93); FR-L3 comprising the amino acid sequence of GVPDRFTGGGSGTDFTLTINTVQAEDAAFFYC (SEQ ID NO: 94); and / or FR-L4 comprising the amino acid sequence of FGTGTKLEIK (SEQ ID NO: 95), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 92-65, such as those possessed by the anti-TIGIT antibody rat6B2.In some cases, the anti-TIGIT antibody comprises an FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 96); an FR-L2 comprising the amino acid sequence of WYQQKPGKSPKLLIY (SEQ ID NO: 97); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC (SEQ ID NO: 99); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 100), or one or more of the foregoing FRs and their combinations with SEQ ID NO: 101. A combination of one or more variants of any of NOs: 96-97 and 99-100 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h6B2.L1H1, h6B2.L1H2, h6B2.L1H3, h6B2.L1H4, and h6B2.L1H5. In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 96); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 98); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC (SEQ ID NO: 99); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 100), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 96 and 98-100, such as those possessed by anti-TIGIT antibodies h6B2.L2H1 and h6B2.L2H5.

[0217] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: comprising X1VX2LKESGPX3X4X5X6PX7X8TLX9LTCX 10 FR-H1 of the amino acid sequence of FSGFSLS (SEQ ID NO: 74), wherein X1 is E or Q; X2 is S or T; X3 is A or G; X4 is I or L; X5 is L or V; X6 is K or Q; X7 is S or T; X8 is H or Q; X9 is S or T; and X 10is S or T; FR-H2 comprising the amino acid sequence of WIRQPX1X2KX3LEWLA (SEQ ID NO: 75), wherein X1 is P or S; X2 is E or G; and X3 is A or G; comprising RLTX1X2KDX3SX4X5QX6X7LX8X9TX 10 X 11 DX 12 X 13 FR-H3 of the amino acid sequence of DTATYYCAH (SEQ ID NO: 76), wherein X1 is I or V; X2 is S or T; X3 is A or T; X4 is K or N; X5 is D or N; X6 is A or V; X7 is F or V; X8 is N or T; X9 is M or V; X 10 Is N or S; X 11 M or V; X 12 is P or T; and X 13is T or V; and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 74-77, such as those possessed by the anti-TIGIT antibody rat6B2 and its derivatives (e.g., h6B2.L1H1, h6B2.L2H1, h6B2.L1H2, h6B2.L1H3, h6B2.L1H4, h6B2.L1H5, and h6B2.L2H5). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVSLKESGPGILQPSHTLSLTCSFSGFSLS (SEQ ID NO: 78); FR-H2 comprising the amino acid sequence of WIRQPSEKGLEWLA (SEQ ID NO: 79); FR-H3 comprising the amino acid sequence of RLTVSKDASNDQAFLNVTSVDTTDTATYYCAH (SEQ ID NO: 80); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 77-80, such as those possessed by the anti-TIGIT antibody rat6B2. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLS (SEQ ID NO: 81); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 82); an FR-H3 comprising the amino acid sequence of RLTVTKDASKNQAVLTMTNMDPVDTATYYCAH (SEQ ID NO: 83); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 77 and 81-83, such as those possessed by anti-TIGIT antibodies h6B2.L1H1 and h6B2.L2H1.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLS (SEQ ID NO: 81); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 82); an FR-H3 comprising the amino acid sequence of RLTITKDASKNQAVLTMTNMDPVDTATYYCAH (SEQ ID NO: 84); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 77, 81-82, and 84, such as possessed by the anti-TIGIT antibody h6B2.L1H2. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLS (SEQ ID NO: 81); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 82); an FR-H3 comprising the amino acid sequence of RLTVTKDTSKNQAVLTMTNMDPVDTATYYCAH (SEQ ID NO: 85); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 77, 81-82, and 85, such as that possessed by the anti-TIGIT antibody h6B2.L1H3.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLS (SEQ ID NO: 81); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 82); an FR-H3 comprising the amino acid sequence of RLTVTKDASKNQVVLTMTNMDPVDTATYYCAH (SEQ ID NO: 86); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 77, 81-82, and 85, such as that possessed by the anti-TIGIT antibody h6B2.L1H4. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLS (SEQ ID NO: 81); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 82); an FR-H3 comprising the amino acid sequence of RLTITKDTSKNQVVLTMTNMDPVDTATYYCAH (SEQ ID NO: 87); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 77), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 77, 81-82, and 87, such as those possessed by anti-TIGIT antibodies h6B2.L1H5 and h6B2.L2H5.

[0218] In another example, the present invention provides an anti-TIGIT antibody comprising at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of TYGMGVS (SEQ ID NO: 110); (b) HVR-H2 comprising the amino acid sequence of SIWWNGNTYYNPSLRS (SEQ ID NO: 111); (c) HVR-H3 comprising the amino acid sequence of TGGAVITWFAY (SEQ ID NO: 112); (d) HVR-L1 comprising the amino acid sequence of KASQSVGKNIA (SEQ ID NO: 113); (e) HVR-L2 comprising the amino acid sequence of YASNRYT (SEQ ID NO: 114); and / or (f) HVR-L3 comprising the amino acid sequence of QHIYNSPYP (SEQ ID NO: 115), or one or more of the above HVRs and their combinations with SEQ ID NO: 116. NO: 110-115 have at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above anti-TIGIT antibodies of the aforementioned embodiments may include, for example, (a) HVR-H1 comprising the amino acid sequence of TYGMGVS (SEQ ID NO: 110); (b) HVR-H2 comprising the amino acid sequence of SIWWNGNTYYNPSLRS (SEQ ID NO: 111); (c) HVR-H3 comprising the amino acid sequence of TGGAVITWFAY (SEQ ID NO: 112); (d) HVR-L1 comprising the amino acid sequence of KASQSVGKNIA (SEQ ID NO: 113); (e) HVR-L2 comprising the amino acid sequence of YASNRYT (SEQ ID NO: 114); and (f) HVR-H3 comprising the amino acid sequence of QHIYNSPYP (SEQ ID NO: 115). NO:115), such as that possessed by the anti-TIGIT antibody rat10A5 and its derivatives (e.g., h10A5.L1H1; h10A5.L2H1; h10A5.L3H1; h10A5.L4H1; h10A5.L1H2; h10A5.L1H3; h10A5.L1H4; and h10A5.L4H4).

[0219] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FR): comprising X1IVMTQSPX2X3X4SX5SX6GX7RX8TX9X 10C (SEQ ID NO: 129), wherein X1 is E or N; X2 is A or K; X3 is S or T; X4 is L or M; X5 is I or V; X6 is I or P; X7 is D or E; X8 is A or V; X9 is L or M; and X 10 is N or S; FR-L2 comprising the amino acid sequence of WYQQKX1GQX2PX3LLIY (SEQ ID NO: 130), wherein X1 is P or T; X2 is A or S; and X3 is Q or R; comprising GX1PX2RFX3GX4GSGTX5FTLTIX6SX7QX8EDX9AX10X 11 FR-L3 of the amino acid sequence of YC (SEQ ID NO: 131), wherein X1 is I or V; X2 is A or D; X3 is S or T; X4 is G or S; X5 is D or E; X6 is N or S; X7 is L or V; X8 is A or S; X9 is A or F; X 10 is F or V; and X 11is F or Y; and / or an FR-L4 comprising the amino acid sequence of FGX1GTKX2EIK (SEQ ID NO: 132), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 129-132, such as those possessed by the anti-TIGIT antibody rat10A5 and its derivatives (e.g., h10A5.L1H1; h10A5.L2H1; h10A5.L3H1; h10A5.L4H1; h10A5.L1H2; h10A5.L1H3; h10A5.L1H4; and h10A5.L4H4). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following light chain variable region FRs: FR-L1 comprising the amino acid sequence of NIVMTQSPKSMSISIGDRVTMNC (SEQ ID NO: 133); FR-L2 comprising the amino acid sequence of WYQQKTGQSPQLLIY (SEQ ID NO: 134); FR-L3 comprising the amino acid sequence of GVPDRFTGGGSGTDFTLTINSVQAEDAAFFYC (SEQ ID NO: 135); and / or FR-L4 comprising the amino acid sequence of FGTGTKLEIK (SEQ ID NO: 136), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 133-136, such as those possessed by the anti-TIGIT antibody rat10A5.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of EIVMTQSPATLSVSPGERATLSC (SEQ ID NO: 137); an FR-L2 comprising the amino acid sequence of WYQQKPGQSPRLLIY (SEQ ID NO: 138); an FR-L3 comprising the amino acid sequence of GVPARFSGSGSGTEFTLTISSLQSEDFAVYYC (SEQ ID NO: 140); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 142), or one or more of the above FRs and their combinations with SEQ ID NO: 143. NO: 137-138, 140, and 142 are any combination of one or more variants having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h10A5.L1H1, h10A5.L1H2, h10A5.L1H3, and h10A5.L1H4. In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of EIVMTQSPATLSVSPGERATLSC (SEQ ID NO: 137); an FR-L2 comprising the amino acid sequence of WYQQKPGQAPRLLIY (SEQ ID NO: 139); an FR-L3 comprising the amino acid sequence of GVPARFSGSGSGTEFTLTISSLQSEDFAVYYC (SEQ ID NO: 140); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 142), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 137, 139-140, and 142, such as that possessed by the anti-TIGIT antibody h10A5.L2H1.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of EIVMTQSPATLSVSPGERATLSC (SEQ ID NO: 137); an FR-L2 comprising the amino acid sequence of WYQQKPGQSPRLLIY (SEQ ID NO: 138); an FR-L3 comprising the amino acid sequence of GIPARFSGSGSGTEFTLTISSLQSEDFAVYYC (SEQ ID NO: 141); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 142), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 137-138 and 141-142, such as possessed by the anti-TIGIT antibody h10A5.L3H1. In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of EIVMTQSPATLSVSPGERATLSC (SEQ ID NO: 137); an FR-L2 comprising the amino acid sequence of WYQQKPGQAPRLLIY (SEQ ID NO: 139); an FR-L3 comprising the amino acid sequence of GIPARFSGSGSGTEFTLTISSLQSEDFAVYYC (SEQ ID NO: 141); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 142), or one or more of the above FRs and their combinations with SEQ ID NO: 143. A combination of one or more variants of any of NOs: 137, 139, and 141-142 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h10A5.L4H1 and h10A5.L4H4.

[0220] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: comprising X1VX2LKESGPX3X4X5X6PX7X8TLX9LTCX 10 FR-H1 of the amino acid sequence of FSGFSLT (SEQ ID NO: 116), wherein X1 is E or Q; X2 is S or T; X3 is A or G; X4 is I or L; X5 is L or V; X6 is K or Q; X7 is S or T; X8 is H or Q; X9 is S or T; and X 10is S or T; FR-H2 comprising the amino acid sequence of WIRQPX1X2KX3LEWLA (SEQ ID NO: 117), wherein X1 is P or S; X2 is E or G; and X3 is A or G; comprising RLTX1X2KDTSX3X4QX5X6LX7X8TX9X 10 DX 11 X 12 FR-H3 of the amino acid sequence of DTATYYCAH (SEQ ID NO: 118), wherein X1 is I or V; X2 is S or T; X3 is K or N; X4 is D or N; X5 is A or V; X6 is F or V; X7 is N or T; X8 is M or V; X9 is N or S; X 10 M or V; X 11 is P or T; and X 12is T or V; and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 116-119, such as those possessed by the anti-TIGIT antibody rat10A5 and its derivatives (e.g., h10A5.L1H1; h10A5.L2H1; h10A5.L3H1; h10A5.L4H1; h10A5.L1H2; h10A5.L1H3; h10A5.L1H4; and h10A5.L4H4). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of QVSLKESGPGILQPSHTLSLTCSFSGFSLT (SEQ ID NO: 120); FR-H2 comprising the amino acid sequence of WIRQPSEKGLEWLA (SEQ ID NO: 121); FR-H3 comprising the amino acid sequence of RLTVSKDTSNDQAFLNVTSVDTTDTATYYCAH (SEQ ID NO: 122); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 119-122, such as those possessed by the anti-TIGIT antibody rat10A5.In some cases, the anti-TIGIT antibody comprises an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLT (SEQ ID NO: 123); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 124); an FR-H3 comprising the amino acid sequence of RLTVTKDTSKNQAVLTMTNMDPVDTATYYCAH (SEQ ID NO: 125); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or one or more of the foregoing FRs and their combinations with SEQ ID NO: 126. A combination of one or more variants of any of NOs: 119 and 123-125 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h10A5.L1H1, h10A5.L2H1, h10A5.L3H1, and h10A5.L4H1. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLT (SEQ ID NO: 123); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 124); an FR-H3 comprising the amino acid sequence of RLTITKDTSKNQAVLTMTNMDPVDTATYYCAH (SEQ ID NO: 126); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 119, 123-124, and 126, such as that possessed by the anti-TIGIT antibody h10A5.L1H2.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLT (SEQ ID NO: 123); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 124); an FR-H3 comprising the amino acid sequence of RLTVTKDTSKNQVVLTMTNMDPVDTATYYCAH (SEQ ID NO: 127); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 119, 123-124, and 127, such as that possessed by the anti-TIGIT antibody h10A5.L1H3. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVTLKESGPALVKPTQTLTLTCTFSGFSLT (SEQ ID NO: 123); an FR-H2 comprising the amino acid sequence of WIRQPPGKALEWLA (SEQ ID NO: 124); an FR-H3 comprising the amino acid sequence of RLTITKDTSKNQVVLTMTNMDPVDTATYYCAH (SEQ ID NO: 128); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 119), or one or more of the above FRs and their combinations with SEQ ID NO: 120. A combination of one or more variants of any of NOs: 119, 123-124, and 128 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h10A5.L1H4 and h10A5.L4H4.

[0221] In another example, the present invention provides an anti-TIGIT antibody comprising at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of EYSIY (SEQ ID NO: 153); (b) HVR-H2 comprising the amino acid sequence of RIDPKNGRTYYVDKFKN (SEQ ID NO: 154); (c) HVR-H3 comprising the amino acid sequence of IYGFYFDF (SEQ ID NO: 155); (d) HVR-L1 comprising the amino acid sequence of KGSQNVNKYLV (SEQ ID NO: 156); (e) HVR-L2 comprising the amino acid sequence of NTDNLQS (SEQ ID NO: 157); and / or (f) HVR-L3 comprising the amino acid sequence of YQYNNGFT (SEQ ID NO: 158), or one or more of the above HVRs and their combinations with SEQ ID NO: 159. NO: 153-158 have at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above anti-TIGIT antibodies of the preceding embodiments may include, for example, (a) HVR-H1 comprising the amino acid sequence of EYSIY (SEQ ID NO: 153); (b) HVR-H2 comprising the amino acid sequence of RIDPKNGRTYYVDKFKN (SEQ ID NO: 154); (c) HVR-H3 comprising the amino acid sequence of IYGFYFDF (SEQ ID NO: 155); (d) HVR-L1 comprising the amino acid sequence of KGSQNVNKYLV (SEQ ID NO: 156); (e) HVR-L2 comprising the amino acid sequence of NTDNLQS (SEQ ID NO: 157); and (f) HVR-H3 comprising the amino acid sequence of YQYNNGFT (SEQ ID NO: 158). NO:158), such as HVR-L3 possessed by the anti-TIGIT antibody rat7E7 and its derivatives (e.g., h7E7.L1H1, h7E7.L2H1, h7E7.L3H1, h7E7.L4H1, h7E7.L5H1, h7E7.L1H2, h7E7.L1H3, h7E7.L1H4, h7E7.L1H5, h7E7.L1H6, h7E7.L1H7, h7E7.L1H8, h7E7.L1H9, h7E7.L5H9, and 7E7.L5aH9a).

[0222] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): a FR-L1 comprising the amino acid sequence of X1IX2LTQSPSX3LSASVGDRVTX4X5C (SEQ ID NO: 180), wherein X1 is D or N; X2 is H or Q; X3 is F or L; X4 is I or L; and X5 is S or T; a FR-L2 comprising the amino acid sequence of WYQQKX1GX2APKLLIY (SEQ ID NO: 181), wherein X1 is L or P; and X2 is E or K; a FR-L3 comprising the amino acid sequence of GX1PSRFSGSGSGTX2X3TLTISSLQPEDX4ATYX5C (SEQ ID NO: 182), wherein X1 is L or P; and X2 is E or K; NO: 182), wherein X1 is I or V; X2 is D or E; X3 is F or Y; X4 is A or F; and X5 is F or Y; and / or an FR-L4 comprising the amino acid sequence of FGX1GTKX2EIK (SEQ ID NO: 183), wherein X1 is Q or S; and X2 is L or V, or one or more of the above FRs and their combinations with SEQ ID NO: A combination of one or more variants of any of NOs: 180-183 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by the anti-TIGIT antibody rat7E7 and its derivatives (e.g., h7E7.L1H1, h7E7.L2H1, h7E7.L3H1, h7E7.L4H1, h7E7.L5H1, h7E7.L1H2, h7E7.L1H3, h7E7.L1H4, h7E7.L1H5, h7E7.L1H6, h7E7.L1H7, h7E7.L1H8, h7E7.L1H9, h7E7.L5H9, and 7E7.L5aH9a).The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following light chain variable region FRs: FR-L1 comprising the amino acid sequence of NIHLTQSPSLLSASVGDRVTLSC (SEQ ID NO: 184); FR-L2 comprising the amino acid sequence of WYQQKLGEAPKLLIY (SEQ ID NO: 185); FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTDYTLTISSLQPEDAATYFC (SEQ ID NO: 186); and / or FR-L4 comprising the amino acid sequence of FGSGTKLEIK (SEQ ID NO: 187), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 184-187, such as those possessed by the anti-TIGIT antibody rat7E7. In some cases, the anti-TIGIT antibody comprises an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEYTLTISSLQPEDFATYFC (SEQ ID NO: 190); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or one or more of the foregoing FRs and their combinations with SEQ ID NO: 197. A combination of one or more variants of any of NOs: 188-190 and 196 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h7E7.L1H1, h7E7.L1H2, h7E7.L1H3, h7E7.L1H4, h7E7.L1H5, h7E7.L1H6, h7E7.L1H7, h7E7.L1H8, and h7E7.L1H9.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEYTLTISSLQPEDFATYFC (SEQ ID NO: 191); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 188-189, 191 and 196, such as that possessed by the anti-TIGIT antibody h7E7.L2H1. In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEFTLTISSLQPEDFATYFC (SEQ ID NO: 192); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 188-189, 192, and 196, such as possessed by the anti-TIGIT antibody h7E7.L3H1.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEYTLTISSLQPEDFATYYC (SEQ ID NO: 193); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 188-189, 193 and 196, such as possessed by the anti-TIGIT antibody h7E7.L4H1. In some cases, the anti-TIGIT antibody comprises an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEFTLTISSLQPEDFATYYC (SEQ ID NO: 194); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or one or more of the foregoing FRs and their combinations with SEQ ID NO: 197. A combination of one or more variants of any of NOs: 188-189, 194 and 196 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h7E7.L5H1 and h7E7.L5H9.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 188); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 189); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEYTLTISSLQPEDFATYYC (SEQ ID NO: 195); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 196), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 188-189 and 195-196, such as possessed by the anti-TIGIT antibody 7E7.L5aH9a.

[0223] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: a FR-H1 comprising the amino acid sequence of EVQLX1QSGX2EX3X4X5PGASVKX6SCKAX7GYTFT (SEQ ID NO: 159), wherein X1 is Q or V; X2 is A or P; X3 is L or V; X4 is K or Q; X5 is K or R; X6 is L or V; and X7 is S or T; a FR-H2 comprising the amino acid sequence of WVX1QX2PX3QX4LEX5X6G (SEQ ID NO: 160), wherein X1 is K or R; X2 is A or R; X3 is G or K; X4 is R or S; X5 is I or W; and X6 is I or M; a FR-H2 comprising the amino acid sequence of RX1TX2TX3DTSX4X5TAYMX6LSSLX7SEDTAX8YX9CX10R (SEQ ID NO: NO: 161), wherein X1 is A or V; X2 is I or L; X3 is A or R; X4 is A or S; X5 is N or S; X6 is E or Q; X7 is R or T; X8 is T or V; X9 is F or Y; and X 10is A or T; and / or a FR-H4 comprising the amino acid sequence of WGQGX1X2VTX3SS (SEQ ID NO: 162), wherein X1 is T or V; X2 is L or M; and X3 is A or V, or one or more of the above FRs and their combinations with SEQ ID NO: NO: 159-162 any one or more variants having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by the anti-TIGIT antibody rat7E7 and its derivatives (e.g., h7E7.L1H1, h7E7.L2H1, h7E7.L3H1, h7E7.L4H1, h7E7.L5H1, h7E7.L1H2, h7E7.L1H3, h7E7.L1H4, h7E7.L1H5, h7E7.L1H6, h7E7.L1H7, h7E7.L1H8, h7E7.L1H9, h7E7.L5H9, and 7E7.L5aH9a). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLQQSGPELQRPGASVKLSCKATGYTFT (SEQ ID NO: 163); FR-H2 comprising the amino acid sequence of WVKQRPKQSLEIIG (SEQ ID NO: 164); FR-H3 comprising the amino acid sequence of RATLTADTSSNTAYMQLSSLTSEDTATYFCTR (SEQ ID NO: 165); and / or FR-H4 comprising the amino acid sequence of WGQGVMVTASS (SEQ ID NO: 166), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 163-166, such as those possessed by the anti-TIGIT antibody rat7E7.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RATLTADTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 172); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or one or more of the above FRs and their combinations with SEQ ID NO: 180. NO: 167-138, 172, and 179 are any combination of one or more variants having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h7E7.L1H1, h7E7.L2H1, h7E7.L3H1, h7E7.L4H1, and h7E7.L5H1. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEWIG (SEQ ID NO: 169); an FR-H3 comprising the amino acid sequence of RATLTADTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 172); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167, 169, 172, and 179, such as that possessed by the anti-TIGIT antibody h7E7.L1H2.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIMG (SEQ ID NO: 170); an FR-H3 comprising the amino acid sequence of RATLTADTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 172); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167, 170, 172, and 179, such as possessed by the anti-TIGIT antibody h7E7.L1H3. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RVTLTADTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 173); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167-168, 173, and 179, such as possessed by the anti-TIGIT antibody h7E7.L1H4.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RATITADTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 174); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167-168, 174, and 179, such as that possessed by the anti-TIGIT antibody h7E7.L1H5. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RATLTRDTSASTAYMELSSLRSEDTAVYFCTR (SEQ ID NO: 175); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167-168, 175, and 179, such as that possessed by the anti-TIGIT antibody h7E7.L1H6.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RATLTADTSASTAYMELSSLRSEDTAVYYCTR (SEQ ID NO: 176); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167-168, 176, and 179, such as that possessed by the anti-TIGIT antibody h7E7.L1H7. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 168); an FR-H3 comprising the amino acid sequence of RATLTADTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 177); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167-168, 177, and 179, such as that possessed by the anti-TIGIT antibody h7E7.L1H8.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEWMG (SEQ ID NO: 171); an FR-H3 comprising the amino acid sequence of RVTITRDTSASTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 178); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or one or more of the above FRs and their combinations with SEQ ID NO: 180. A combination of one or more variants of any of NOs: 167, 171, and 178-179 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h7E7.L1H9 and h7E7.L5H9. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 167); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIMG (SEQ ID NO: 170); an FR-H3 comprising the amino acid sequence of RVTITRDTSASTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 178); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 179), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 167, 170, and 178-179, such as possessed by the anti-TIGIT antibody 7E7.L5aH9a.

[0224] In another example, the present invention provides an anti-TIGIT antibody comprising at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 comprising the amino acid sequence of EHSIY (SEQ ID NO: 215); (b) HVR-H2 comprising the amino acid sequence of RIDPKNGRTYFVDKFKN (SEQ ID NO: 216); (c) HVR-H3 comprising the amino acid sequence of IDGFYFDF (SEQ ID NO: 217); (d) HVR-L1 comprising the amino acid sequence of KGSQNVNKYLV (SEQ ID NO: 218); (e) HVR-L2 comprising the amino acid sequence of STDNLQS (SEQ ID NO: 219); and / or (f) HVR-L3 comprising the amino acid sequence of YQYNNGFT (SEQ ID NO: 220), or one or more of the above HVRs and their combinations with SEQ ID NO: 221. NO: 215-220 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, any of the above-mentioned anti-TIGIT antibodies of the preceding embodiments may include, for example, (a) HVR-H1 comprising the amino acid sequence of EHSIY (SEQ ID NO: 215); (b) HVR-H2 comprising the amino acid sequence of RIDPKNGRTYFVDKFKN (SEQ ID NO: 216); (c) HVR-H3 comprising the amino acid sequence of IDGFYFDF (SEQ ID NO: 217); (d) HVR-L1 comprising the amino acid sequence of KGSQNVNKYLV (SEQ ID NO: 218); (e) HVR-L2 comprising the amino acid sequence of STDNLQS (SEQ ID NO: 219); and (f) HVR-H3 comprising the amino acid sequence of YQYNNGFT (SEQ ID NO: 220). NO: 220), such as HVR-L3 possessed by the anti-TIGIT antibody rat15C8 and its derivatives (e.g., h15C8.L1H1, h15C8.L2H1, h15C8.L3H1, h15C8.L4H1, h15C8.L5H1, h15C8.L1H2, h15C8.L1H3, h15C8.L1H4, h15C8.L1H5, h15C8.L1H6, h15C8.L1H7, h15C8.L1H8, h15C8.L1H9, h15C8.L5H9, and 5C8.L5aH9a).

[0225] In some cases, the antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FRs): a FR-L1 comprising the amino acid sequence of X1IX2LTQSPSX3LSASVGDRVTX4X5C (SEQ ID NO: 243), wherein X1 is D or N; X2 is H or Q; X3 is F or L; X4 is I or L; and X5 is S or T; a FR-L2 comprising the amino acid sequence of WYQQKX1GX2APKLLIY (SEQ ID NO: 244), wherein X1 is L or P and X2 is E or K; a FR-L3 comprising the amino acid sequence of GX1PSRFSGSGSGTX2X3TLTISSLQPEDX4ATYX5C (SEQ ID NO: 245), wherein X1 is D or N; X2 is H or Q; X3 is F or L; X4 is I or L; and X5 is S or T; NO: 245), wherein X1 is I or V; X2 is D or E; X3 is F or Y; X4 is A or F; and X5 is F or Y; and / or an FR-L4 comprising the amino acid sequence of FGX1GTKX2EIK (SEQ ID NO: 246), wherein X1 is Q or S and X2 is L or V, or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 243-246, such as those obtained from the anti-TIGIT antibody rat15C8 and derivatives thereof (e.g., h15C8.L1H1, h15C8.L2H1, h15C8.L3 H1, h15C8.L4H1, h15C8.L5H1, h15C8.L1H2, h15C8.L1H3, h15C8.L1H4, h15C8.L1H5, h15C8.L1H6, h15C8.L1H7, h15C8.L1H8, h15C8.L1H9, h15C8.L5H9, and 5C8.L5aH9a).The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following light chain variable region FRs: FR-L1 comprising the amino acid sequence of NIHLTQSPSLLSASVGDRVTLSC (SEQ ID NO: 247); FR-L2 comprising the amino acid sequence of WYQQKLGEAPKLLIY (SEQ ID NO: 248); FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTDYTLTISSLQPEDAATYFC (SEQ ID NO: 249); and / or FR-L4 comprising the amino acid sequence of FGSGTKLEIK (SEQ ID NO: 250), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 247-250, such as those possessed by the anti-TIGIT antibody rat15C8. In some cases, the anti-TIGIT antibody comprises an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEYTLTISSLQPEDFATYFC (SEQ ID NO: 253); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or one or more of the foregoing FRs and their combinations with SEQ ID NO: 260. A combination of one or more variants of any of NOs: 251-253 and 259 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h15C8.L1H1, h15C8.L1H2, h15C8.L1H3, h15C8.L1H4, h15C8.L1H5, h15C8.L1H6, h15C8.L1H7, h15C8.L1H8, and h15C8.L1H9.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEYTLTISSLQPEDFATYFC (SEQ ID NO: 254); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 251-252, 254, and 259, such as possessed by the anti-TIGIT antibody h15C8.L2H1. In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEFTLTISSLQPEDFATYFC (SEQ ID NO: 255); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 251-252, 255, and 259, such as that possessed by the anti-TIGIT antibody h15C8.L3H1.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GIPSRFSGSGSGTEYTLTISSLQPEDFATYYC (SEQ ID NO: 256); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 251-252, 256, and 259, such as that possessed by the anti-TIGIT antibody h15C8.L4H1. In some cases, the anti-TIGIT antibody comprises an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEFTLTISSLQPEDFATYYC (SEQ ID NO: 257); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or one or more of the above FRs and their combinations with SEQ ID NO: 260. A combination of one or more variants having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of IDNOs: 251-252, 257, and 259, such as those possessed by anti-TIGIT antibodies h15C8.L5H1 and h15C8.L5H9.In some cases, the anti-TIGIT antibody includes an FR-L1 comprising the amino acid sequence of DIQLTQSPSFLSASVGDRVTITC (SEQ ID NO: 251); an FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 252); an FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTEYTLTISSLQPEDFATYYC (SEQ ID NO: 258); and / or an FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 259), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 251-252, and 258-259, such as possessed by the anti-TIGIT antibody 5C8.L5aH9a.

[0226] In some cases, the antibody further comprises at least one, two, three, or four of the following heavy chain variable region FRs: an FR-H1 comprising the amino acid sequence of EVQLX1QSGX2EX3X4X5PGASVKX6SCKASGYTFT (SEQ ID NO: 221), wherein X1 is Q or V; X2 is A or P; X3 is L or V; X4 is K or Q; X5 is K or R; and X6 is L or V; an FR-H2 comprising the amino acid sequence of WX1X2QX3PX4QX5LEX6X7G (SEQ ID NO: 222), wherein X1 is L or V; X2 is K or R; X3 is A or R; X4 is G or K; X5 is R or S; X6 is I or W; and X7 is I or M; an FR-H2 comprising the amino acid sequence of RX1TX2TX3X4TSX5X6TAYMX7LSSLX8SEDTAX9YX (SEQ ID NO: 223), wherein X1 is L or V; X2 is K or R; X3 is A or R; X4 is G or K; X5 is R or S; X6 is I or W; and X7 is I or M; 10 FR-H3 of the amino acid sequence of CAR (SEQ ID NO: 223), wherein X1 is A or V; X2 is I or L; X3 is R or T; X4 is D or N; X5 is A or S; X6 is N or S; X7 is E or Q; X8 is R or T; X9 is I or V; and X 10is F or Y; and / or FR-H4 comprising the amino acid sequence of WGQGX1X2VTX3SS (SEQ ID NO: 224), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 221-224, such as those obtained from the anti-TIGIT antibody rat15C8 and its derivatives (e.g., h15C8.L1H1, h15C8.L2H1, h15C8.L3 H1, h15C8.L4H1, h15C8.L5H1, h15C8.L1H2, h15C8.L1H3, h15C8.L1H4, h15C8.L1H5, h15C8.L1H6, h15C8.L1H7, h15C8.L1H8, h15C8.L1H9, h15C8.L5H9, and 5C8.L5aH9a). The anti-TIGIT antibody may further include, for example, at least one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of EVQLQQSGPELQRPGASVKLSCKASGYTFT (SEQ ID NO: 225); FR-H2 comprising the amino acid sequence of WLKQRPKQSLEIIG (SEQ ID NO: 226); FR-H3 comprising the amino acid sequence of RATLTTNTSSNTAYMQLSSLTSEDTAIYFCAR (SEQ ID NO: 227); and / or FR-H4 comprising the amino acid sequence of WGQGVMVTASS (SEQ ID NO: 228), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 225-228, such as those possessed by the anti-TIGIT antibody rat15C8.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIIG (SEQ ID NO: 230); an FR-H3 comprising the amino acid sequence of RATLTTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 235); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or one or more of the above FRs and their combinations with SEQ ID NO: 243. NO: 229-230, 235, and 242 are any combination of one or more variants having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h15C8.L1H1, h15C8.L2H1, h15C8.L3H1, h15C8.L4H1, and h15C8.L5H1. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEIIG (SEQ ID NO: 231); an FR-H3 comprising the amino acid sequence of RATLTTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 235); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229, 231, 235, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H2.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEWIG (SEQ ID NO: 232); an FR-H3 comprising the amino acid sequence of RATLTTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 235); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229, 232, 235, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H3. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIMG (SEQ ID NO: 233); an FR-H3 comprising the amino acid sequence of RATLTTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 235); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229, 233, 235, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H4.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIIG (SEQ ID NO: 230); an FR-H3 comprising the amino acid sequence of RVTLTTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 236); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229-230, 236, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H5. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIIG (SEQ ID NO: 230); an FR-H3 comprising the amino acid sequence of RATITTDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 237); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229-230, 237, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H6.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIIG (SEQ ID NO: 230); an FR-H3 comprising the amino acid sequence of RATLTRDTSASTAYMELSSLRSEDTAVYFCAR (SEQ ID NO: 238); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229-230, 238, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H7. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIIG (SEQ ID NO: 230); an FR-H3 comprising the amino acid sequence of RATLTTDTSASTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 239); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229-230, 239, and 242, such as that possessed by the anti-TIGIT antibody h15C8.L1H8.In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WVRQAPGQRLEWMG (SEQ ID NO: 234); an FR-H3 comprising the amino acid sequence of RVTITRDTSASTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 240); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or one or more of the above FRs and their combinations with SEQ ID NO: 243. A combination of one or more variants of any of NO: 229, 234, 240, and 242 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity), such as those possessed by anti-TIGIT antibodies h15C8.L1H9 and h15C8.L5H9. In some cases, the anti-TIGIT antibody includes an FR-H1 comprising the amino acid sequence of EVQLVQSGAEVKKPGASVKVSCKASGYTFT (SEQ ID NO: 229); an FR-H2 comprising the amino acid sequence of WLRQAPGQRLEIMG (SEQ ID NO: 233); an FR-H3 comprising the amino acid sequence of RATITTDTSASTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 241); and / or an FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 242), or a combination of one or more of the foregoing FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 229, 233, and 241-242, such as that possessed by the anti-TIGIT antibody 5C8.L5aH9a.

[0227] Also provided are anti-TIGIT antibodies comprising a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 60, 62, 66, 101, 103, 104, 105, 106, 107, 145, 146, 147, 148, 197, 199, 200, 201, 202, 203, 204, 205, 206, 207, 213, 260, 262, 263, 264, 265, 266, 267, 268, 269, 270, or 276, or a sequence thereof. Also provided are anti-TIGIT antibodies comprising a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 61, 63, 64, 67, 102, 108, 109, 144, 149, 150, 151, 152, 198, 208, 209, 210, 211, 212, 214, 261, 271, 272, 273, 274, 275, or 277, or a sequence thereof.

[0228] In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 60, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 61, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 62, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 63, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 62, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 64, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 66, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 67, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 101, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 102, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 103, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 108, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 104, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 108, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 105, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 108, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 106, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 108, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 107, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 109, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 143, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 144, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 145, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 149, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 145, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 150, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 145, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 151, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 145, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 152, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 147, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 149, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 148, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 149, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 148, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 152, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 197, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 198, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 199, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 200, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 201, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 202, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 203, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 204, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 205, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 206, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 207, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 208, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 207, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 212, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 199, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 209, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 199, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 210, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 199, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 211, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 199, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 212, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 213, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 214, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 260, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 261, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 262, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 263, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 264, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 265, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 266, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 267, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 268, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 269, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 270, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 271, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 270, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 275, or its sequence.In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 262, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 272, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 262, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 273, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 262, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 274, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 262, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 275, or its sequence. In some cases, the anti-TIGIT antibody comprises a heavy chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 276, or its sequence, and a light chain variable region having at least 90% (e.g., 91%, 92%, 93%, or 94%), or at least 95% (e.g., 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 277, or its sequence.

[0229] In another aspect, an anti-TIGIT antibody is provided, wherein the antibody comprises the VH of any of the embodiments provided above and the VL of any of the embodiments provided above, wherein one or both of the variable domain sequences include a post-translational modification.

[0230] In some cases, in addition to human TIGIT, any of the anti-TIGIT antibodies described above may be able to bind to rabbit TIGIT. In some cases, any of the anti-TIGIT antibodies described above may be able to bind to both human TIGIT and cynomolgus monkey TIGIT. In some cases, any of the anti-TIGIT antibodies described above may be able to bind to human TIGIT, cynomolgus monkey TIGIT, and rabbit TIGIT. In some cases, any of the anti-TIGIT antibodies described above (e.g., 4.1D3 or a derivative thereof) may be able to bind to human TIGIT, cynomolgus monkey TIGIT, and rabbit TIGIT, but not mouse TIGIT.

[0231] In some cases, the anti-TIGIT antibody binds to human TIGIT with a Kd of about 10 nM or less and binds to cynomolgus monkey TIGIT with a Kd of about 10 nM or less (e.g., binds to human TIGIT with a Kd of about 0.1 nM to about 1 nM and binds to cynomolgus monkey TIGIT with a Kd of about 0.5 nM to about 1 nM, e.g., binds to human TIGIT with a Kd of about 0.1 nM or less and binds to cynomolgus monkey TIGIT with a Kd of about 0.5 nM or less).

[0232] In some embodiments, the anti-TIGIT antibody is an antagonist antibody. The antagonist antibody can specifically bind to TIGIT and inhibit or block the interaction of TIGIT with the poliovirus receptor (PVR) (for example, the antagonist antibody inhibits intracellular signaling mediated by TIGIT binding to PVR). In some cases, the antagonist antibody inhibits or blocks human TIGIT binding to human PVR with an IC50 value of 10nM or lower (for example, 1nM to about 10nM). In some cases, the antagonist antibody inhibits or blocks cynomolgus monkey TIGIT binding to cynomolgus monkey PVR with an IC50 value of 50nM or lower (for example, 1nM to about 50nM, for example, 1nM to about 5nM).

[0233] In other embodiments, the anti-TIGIT antibody can be an agonist antibody. The agonist antibody can specifically bind to TIGIT and stimulate the interaction of PVR with CD226 or CD96. For example, the agonist antibody can specifically bind to TIGIT and stimulate the interaction of PVR with CD226 and CD96 (e.g., bind to human TIGIT and stimulate the interaction of human PVR with human CD226 and human CD96 and / or bind to cynomolgus monkey TIGIT and stimulate the interaction of cynomolgus monkey PVR with cynomolgus monkey CD226 and cynomolgus monkey CD96).

[0234] In yet another aspect, the present invention provides an isolated antibody that competes for binding to TIGIT with any of the anti-TIGIT antibodies described above. In yet another aspect, the present invention provides an isolated antibody that binds to the same epitope as the anti-TIGIT antibodies described above.

[0235] The anti-TIGIT antibody according to any of the above embodiments is a monoclonal antibody, including chimeric, humanized, or human antibodies. In one embodiment, the anti-TIGIT antibody is an antibody fragment, such as an Fv, Fab, Fab', scFv, diabody, or F(ab')2 fragment. In another embodiment, the antibody is a full-length antibody, such as a complete IgG antibody (e.g., a complete IgG1 antibody) or other antibody class or isotype, as defined herein.

[0236] In yet another aspect, an anti-TIGIT antibody according to any of the above embodiments may incorporate any of the features described in Sections 1-7 below, singly or in combination:

[0237] 1. Antibody affinity

[0238] In certain embodiments, the antibodies provided herein (e.g., anti-TIGIT antibodies) have a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10 -8 M or less, such as 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 M).

[0239] In one embodiment, Kd is measured by radiolabeled antigen binding assay (RIA). In one embodiment, RIA is performed using a Fab version of the antibody of interest and its antigen. For example, by titrating the antibody with a minimal concentration of ( 1251) Label the antigen to equilibrate the Fab, and then use an anti-Fab antibody coated plate to capture the bound antigen to measure the solution binding affinity of the Fab for the antigen (see, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999)). To establish the conditions for the assay, Multiwell plates (Thermo Scientific) were coated overnight with 5 μg / ml capture anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6) and then blocked with 2% (w / v) bovine serum albumin in PBS at room temperature (approximately 23°C) for 2-5 hours. In non-adsorbent plates (Nunc #269620), 100 pM or 26 pM [ 125 The antigen is mixed with serially diluted Fab of interest (e.g., consistent with the evaluation of the anti-VEGF antibody, Fab-12, in Presta et al., Cancer Res. 57:4593-4599 (1997)). The Fab of interest is then incubated overnight; however, the incubation can be continued for a longer period (e.g., about 65 hours) to ensure that equilibrium is reached. Thereafter, the mixture is transferred to a capture plate and incubated at room temperature (e.g., 1 hour). The solution is then removed and the plate is washed with 0.1% polysorbate 20 in PBS. Wash the plate 8 times. After the plate is dry, add 150 μl / well scintillation fluid (MICROSCINT-20 TM ; Packard), then in TOPCOUNT TM Plates were counted for 10 minutes on a gamma counter (Packard).Concentrations of each Fab that gave less than or equal to 20% of maximal binding were selected for competitive binding assays.

[0240] According to another embodiment, Kd is obtained using For example, at 25°C using an immobilized antigen CM5 chip at approximately 10 response units (RU) -2000 or The assay is performed using a BIAcore BioSensor ® -3000 (BIAcore, Inc., Piscataway, NJ). In one embodiment, a carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) is activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. Antigen is diluted to 5 μg / ml (approximately 0.2 μM) with 10 mM sodium acetate, pH 4.8, and then injected at a flow rate of 5 μl / min to obtain approximately 10 response units (RU) of coupled protein. Following antigen injection, 1 M ethanolamine is injected to block unreacted groups. For kinetic measurements, a 5% ethanolamine solution containing 0.05% polysorbate 20 (TWEEN-20) is injected at 25°C at a flow rate of approximately 25 μl / min. TM ) surfactant in PBS (PBST) two-fold serial dilution of Fab (0.78nM to 500nM). Using a simple one-to-one Langmuir binding model ( Evaluation Software version 3.2) by simultaneously fitting the association and dissociation sensorgrams to calculate the association rate (k on ) and dissociation rate (k off ). With ratio k off / k on Calculate the equilibrium dissociation constant (Kd). See, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). If the association rate exceeds 10 according to the surface plasmon resonance assay above, 6 M -1 S -1 , the binding rate can be measured using the fluorescence quenching technique, i.e., on a spectrometer such as a spectrophotometer equipped with a cut-off flow device (Aviv Instruments) or a 8000 series SLM-AMINCO TM The increase or decrease in fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm bandpass) of 20 nM anti-antigen antibody (Fab form) in PBS pH 7.2 at 25°C was measured in a spectrophotometer (ThermoSpectronic) using a stirred cuvette in the presence of increasing concentrations of antigen.

[0241] 2. Antibody fragments

[0242] In certain embodiments, the antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments, and other fragments described below. For a review of certain antibody fragments, see Hudson et al., Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, for example, Pluckthün, in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds. (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments that contain salvage receptor binding epitope residues and have extended in vivo half-lives, see U.S. Pat. No. 5,869,046.

[0243] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0244] Single-domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody.In certain embodiments, the single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516 Bl).

[0245] Antibody fragments can be produced by a variety of techniques, including but not limited to proteolytic digestion of intact antibodies and production in recombinant host cells (eg, E. coli or phage), as described herein.

[0246] 3. Chimeric and humanized antibodies

[0247] In certain embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described in, for example, U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81: 6851-6855 (1984). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In another example, a chimeric antibody is a "class-switched" antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0248] In certain embodiments, chimeric antibodies are humanized antibodies. Generally, non-human antibodies are humanized to reduce immunogenicity to people while retaining the specificity and affinity of the parent non-human antibody. Generally, humanized antibodies include one or more variable domains, wherein HVR, such as CDR (or part thereof) are derived from non-human antibodies, and FR (or part thereof) are derived from human antibody sequences. Optionally, humanized antibodies also include at least a portion of human constant region. In some embodiments, some FR residues in humanized antibodies are replaced with corresponding residues from non-human antibodies (such as antibodies derived from HVR residues), for example, to restore or improve antibody specificity or affinity.

[0249] Humanized antibodies and methods for their generation are reviewed in, for example, Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described in, for example, Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method of FR shuffling).

[0250] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best-fit" method (see, e.g., Sims et al., J. Immunol. 151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al., J. Immunol., 151:262-264). 3 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived by screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0251] 4. Human Antibodies

[0252] In certain embodiments, the antibodies provided herein are human antibodies. Human antibodies can be generated using a variety of techniques known in the art. Generally, human antibodies are described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[0253] Human antibodies can be prepared by administering immunogens to transgenic animals that have been modified to generate complete human antibodies or complete antibodies with human variable regions in response to antigenic attack. Such animals typically contain all or part of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or which exist extrachromosomally or are randomly integrated into the chromosomes of the animal. In such transgenic mice, the endogenous immunoglobulin loci have generally been inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23: 1117-1125 (2005). Also visible, for example, are U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE. TM technology; U.S. Patent No. 5,770,429, which describes technology; U.S. Patent No. 7,041,870, which describes KM technology, and U.S. Patent Application Publication No. US 2007 / 0061900, which describes Technology). The human variable regions from intact antibodies produced by such animals can be further modified, for example, by combining with a different human constant region.

[0254] Human antibodies can also be generated by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for generating human monoclonal antibodies have been described (see, for example, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991)). Human antibodies generated via human B cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Other methods include those described, for example, in U.S. Pat. No. 7,189,826 (which describes the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (which describes human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3):185-91 (2005).

[0255] It is also possible to generate human antibodies by separating the Fv clone variable domain sequence selected from the phage display library derived from humans.Then, such variable domain sequence can be combined with the desired human constant domain.The technology of selecting human antibodies from the antibody library is described below.

[0256] 5. Library-derived Antibodies

[0257] The antibodies of the present invention can be isolated by screening combinatorial libraries for antibodies with the desired one or more activities. For example, a variety of methods for generating phage display libraries and screening such libraries for antibodies with the desired binding characteristics are known in the art. Such methods are reviewed in, for example, Hoogenboom et al., in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001), and further described in, for example, McCafferty et al., Nature 348: 552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248: 161-175 (Lo et al., ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J.Mol.Biol.338(2):299-310(2004); Lee et al., J.Mol.Biol.340(5):1073-1093(2004); Fellouse, Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004); and Lee et al., J. Immunol. Methods 284(1-2):119-132(2004).

[0258] In certain phage display methods, repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR) and randomly recombined in phage libraries, which can then be screened for antigen-binding phage, as described in Winter et al., Ann. Rev. Immunol., 12:433-455 (1994). Phage typically display antibody fragments as single-chain Fv (scFv) fragments or as Fab fragments. Libraries from immunized sources provide high-affinity antibodies to the immunogen without the need to construct hybridomas. Alternatively, a natural repertoire can be cloned (e.g., from humans) to provide a single source of antibodies to a large number of non-self and also self-antigens in the absence of any immunization, as described by Griffiths et al., EMBO J, 12:725-734 (1993). Finally, unimmunized libraries can also be generated synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences to encode highly variable CDR3 regions and achieve rearrangement in vitro, as described by Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373, and U.S. Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0259] Antibodies or antibody fragments isolated from human antibody libraries are considered human antibodies or human antibody fragments herein.

[0260] 6. Multispecific Antibodies

[0261] In any of the above aspects, the anti-TIGIT antibodies provided herein (e.g., 4.1D3 or a variant thereof, e.g., 4.1D3.Q1E) may be multispecific antibodies, e.g., bispecific antibodies. Multispecific antibodies may be monoclonal antibodies having binding specificity for at least two different sites. In certain embodiments, bispecific antibodies may bind to two different epitopes of TIGIT. In certain embodiments, one of the binding specificities is for TIGIT and the other is for any other antigen (e.g., a second biological molecule, e.g., a cell surface antigen, e.g., a tumor antigen). Thus, bispecific anti-TIGIT antibodies may have binding specificity for TIGIT and a second biological molecule, such as PD-1, PD-L1, PD-L2, OX40, PD-1, CTLA-4, LAG3, TIM3, BTLA, VISTA, B7H4, or CD96. Bispecific antibodies may also be prepared as full-length antibodies or antibody fragments.

[0262] In other embodiments, the bispecific antibody can bind to two different epitopes of TIGIT, OX40, PD-1, PD-L1, PD-L2. In certain embodiments, one of the binding specificities is for TIGIT and the other is for any other antigen (e.g., a second biological molecule, such as OX40). In other embodiments, the bispecific antibody can have binding specificities for TIGIT and PD-L1; TIGIT and PD-L2; TIGIT and PD-1; TIGIT and CTLA-4; TIGIT and LAG3; TIGIT and TIM3; TIGIT and BTLA; TIGIT and VISTA; TIGIT and B7H4; or TIGIT and CD96, wherein the bispecific antibody is preferably an antagonist antibody for TIGIT and an antagonist antibody for its second target. In other embodiments, the bispecific antibody can have binding specificities for TIGIT and CD226; TIGIT and CD28; TIGIT and CD27; TIGIT and CD137; TIGIT and HVEM; TIGIT and GITR; TIGIT and MICA; TIGIT and ICOS; TIGIT and NKG2D; or TIGIT and 2B4, wherein the bispecific antibody is preferably an antagonist antibody for TIGIT and its second target. In other embodiments, the bispecific antibody can have a binding specificity for TIGIT that is not antagonistic in nature (i.e., the bispecific antibody does not act as a TIGIT antagonist).

[0263] Techniques for generating multispecific antibodies include, but are not limited to, recombinant co-expression of two pairs of immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, Nature 305:537 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J. 10:3655 (1991)), and "knob-into-hole" engineering (see, e.g., U.S. Patent No. 5,731,168). The "knob-into-hole" engineering of multispecific antibodies can be used to generate a first arm containing a knob and a second arm containing a hole, wherein the knob of the first arm can be incorporated into the hole of the second arm. In one embodiment, the knob of the multispecific antibody of the present invention can be an anti-TIGIT arm. Alternatively, in one embodiment, the knob of the multispecific antibody of the present invention can be an anti-TIGIT arm. Multispecific antibodies can also be engineered using immunoglobulin crossover / exchange (also known as Fab domain exchange or CrossMab format) technology (see, for example, WO 2009 / 080253; Schaefer et al., Proc. Natl. Acad. Sci. USA, 108: 11187-11192 (2011)). It is also possible to engineer multispecific antibodies by engineering electrostatic manipulation effects for generating antibody Fc-heterodimer molecules (WO 2009 / 089004A1); crosslinking two or more antibodies or fragments (see, for example, U.S. Patent No. 4,676,980, and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to generate bispecific antibodies (see, for example, Kostelny et al., J. Immunol., 148 (5): 1547-1553 (1992)); using antibodies for generating Multispecific antibodies can be generated by using the "diabody" technology to generate bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and by preparing trispecific antibodies as described, e.g., in Tutt et al., J. Immunol. 147:60 (1991).

[0264] Also included herein are engineered antibodies having three or more functional antigen binding sites, including "octopus antibodies" (see, e.g., US 2006 / 0025576A1).

[0265] Antibodies or antibody fragments thereof may also include "dual-acting FAbs" or "DAFs" that comprise antigen binding sites that bind to TIGIT and another, different antigen (eg, a second biological molecule) (see, eg, US 2008 / 0069820).

[0266] 7. Antibody variants

[0267] In certain embodiments, amino acid sequence variants of the anti-TIGIT antibodies of the present invention are encompassed. As described in detail herein, anti-TIGIT antibodies can be optimized based on desired structural and functional properties. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions, and / or insertions and / or substitutions of residues within the amino acid sequence of the antibody. Any combination of deletions, insertions, and substitutions can be performed to obtain the final construct, as long as the final construct possesses the desired characteristics, for example, antigen binding.

[0268] I. Substitution, Insertion, and Deletion Variants

[0269] In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites of interest for substitution mutagenesis include HVRs and FRs. Conservative substitutions are shown in Table 1 under the heading "Preferred Substitutions." More substantial changes are provided in Table 1 under the heading "Exemplary Substitutions," and are further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest, and the product screened for desired activity, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0270] Table 1: Exemplary and preferred amino acid substitutions

[0271]

[0272]

[0273] According to common side chain properties, amino acids can be grouped as follows:

[0274] (1) Hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

[0275] (2) Neutral, hydrophilic: Cys, Ser, Thr, Asn, Gln;

[0276] (3) Acidic: Asp, Glu;

[0277] (4) Basic: His, Lys, Arg;

[0278] (5) Residues that affect chain orientation: Gly, Pro;

[0279] (6) Aromatic: Trp, Tyr, Phe.

[0280] Non-conservative substitutions will entail exchanging a member of one of these classes for a member of another class.

[0281] A class of substitution variants involves replacing one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Generally, the resulting variants selected for further study will have changes (e.g., improvements) in certain biological properties relative to the parent antibody (e.g., increased affinity, reduced immunogenicity) and / or will substantially retain certain biological properties of the parent antibody. Exemplary substitution variants are affinity-matured antibodies, which can be conveniently generated, for example, using affinity maturation techniques based on phage display, such as those described herein. In short, one or more HVR residues are mutated, and the variant antibodies are displayed on phage and screened for specific biological activity (e.g., binding affinity).

[0282] HVR can be changed (e.g., replaced), for example to improve antibody affinity. Such changes can be made to HVR "hot spots", i.e., residues encoded by codons that undergo mutations at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207: 179-196 (2008)), and / or residues contacting the antigen, wherein the resulting variant VH or VL is tested for binding affinity. Affinity maturation by construction and reselection of secondary libraries has been described in, e.g., Hoogenboom et al., in Methods in Molecular Biology 178: 1-37 (O'Brien et al., Human Press, Totowa, NJ, (2001)). In some embodiments of affinity maturation, diversity is introduced into the variable genes selected for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-guided mutagenesis). Then, a secondary library is created. The library is then screened to identify any antibody variants with the desired affinity. Another approach to introducing diversity involves an HVR-guided approach in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding can be specifically identified, for example, using alanine scanning mutagenesis or modeling. In particular, CDR-H3 and CDR-L3 are often targeted.

[0283] In certain embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, as long as such changes do not substantially reduce the ability of the antibody to bind to antigen. For example, conservative changes (e.g., conservative substitutions, as provided herein) may be made to HVRs that do not substantially reduce binding affinity. For example, such changes may be outside the antigen contact residues in the HVRs. In certain embodiments of the variant VH and VL sequences provided above, each HVR is unchanged or contains no more than 1, 2, or 3 amino acid substitutions.

[0284] A method for identifying residues or regions that can be used as mutagenesis targets in antibodies is called "alanine scanning mutagenesis," as described by Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, groups of residues or target residues (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction between the antibody and the antigen is affected. Further substitutions can be introduced at amino acid positions that demonstrate functional sensitivity to the initial substitutions. Alternatively or in addition, the crystal structure of the antigen-antibody complex is used to identify contact points between the antibody and the antigen. As candidates for substitution, such contact residues and adjacent residues can be targeted or eliminated. Variants can be screened to determine whether they contain desired properties.

[0285] Amino acid sequence insertions include amino and / or carboxyl terminal fusions ranging in length from 1 residue to polypeptides containing 100 or more residues, and intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertion variants of the antibody molecule include fusions of the N- or C-terminus of the antibody with an enzyme (e.g., for ADEPT) or a polypeptide that extends the serum half-life of the antibody.

[0286] II. Glycosylation variants

[0287] In certain embodiments, the anti-TIGIT antibodies of the invention (e.g., 4.1D3 or a variant thereof, e.g., 4.1D3.Q1E) can be modified to increase or decrease the degree of antibody glycosylation. Addition or deletion of glycosylation sites to the anti-TIGIT antibodies of the invention can be conveniently achieved by altering the amino acid sequence to create or eliminate one or more glycosylation sites.

[0288] In the case where the antibody comprises an Fc district, the carbohydrate to which it is attached can be changed. The natural antibody produced by mammalian cells generally comprises branched, biantennary oligosaccharides, which are generally attached to the Asn297 of the CH2 domains in the Fc district by N connection. See, for example, Wright et al., TIBTECH 15:26-32 (1997). Oligosaccharides can include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as the fucose attached to the GlcNAc in the biantennary oligosaccharide structure " backbone ". In some embodiments, the oligosaccharides in the antibodies of the present invention can be modified to create antibody variants with certain improved characteristics.

[0289] In one embodiment, an anti-TIGIT antibody variant is provided, which has a carbohydrate structure lacking fucose attached (directly or indirectly) to the Fc region. For example, the amount of fucose in such an antibody can be 1% to 80%, 1% to 65%, 5% to 65% or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297 relative to the sum of all sugar structures (e.g., complex, hybrid and high mannose structures) attached to Asn297, as measured by MALDI-TOF mass spectrometry, for example as described in WO 2008 / 077546. Asn297 refers to an asparagine residue located at approximately position 297 (EU numbering of Fc region residues) in the Fc region; however, Asn297 may also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in the antibody. Such fucosylated variants may have improved ADCC function. See, eg, US Patent Publication Nos. US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications relating to "defucosylated" or "fucose-deficient" antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol. 336: 1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004).Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells deficient in protein fucosylation (Ripka et al., Arch. Biochem. Biophys. 249: 533-545 (1986); U.S. Patent Application No. US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., particularly in Example 11), and knockout cell lines, such as α-1,6-fucosyltransferase gene FUT8 knockout CHO cells (see, e.g., Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4): 680-688 (2006); and WO 2003 / 085107).

[0290] Further provided are anti-TIGIT antibody variants having bisected oligosaccharides, for example, biantennary oligosaccharides attached to the Fc region of the antibody are bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described in, for example, WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described in, for example, WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0291] III. Fc Region Variants

[0292] In certain embodiments, one or more amino acid modifications can be introduced into the Fc region of an anti-TIGIT antibody of the invention (e.g., 4.1D3 or a variant thereof, e.g., 4.1D3.Q1E), thereby generating an Fc region variant (see, e.g., US 2012 / 0251531). The Fc region variant can comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) compri...

Claims

1. An antibody that specifically binds to human TIGIT, wherein the antibody comprises a heavy chain variable region (VH) as shown in the amino acid sequence of SEQ ID NO: 34 and a light chain variable region (VL) as shown in the amino acid sequence of SEQ ID NO: 36, wherein the antibody is an IgG class antibody.

2. The antibody of claim 1, wherein the antibody is a monoclonal antibody.

3. The antibody of claim 1 or 2, wherein the antibody is a human antibody.

4. The antibody of claim 1 or 2, wherein the antibody is a full-length antibody.

5. The antibody of claim 1 or 2, wherein the antibody is an IgG1 subclass antibody.

6. The antibody of claim 1 or 2, wherein the antibody comprises a human IgG1 Fc region.

7. The antibody of claim 6, wherein the human IgG1 Fc region comprises one or more amino acid modifications.

8. The antibody of claim 1 or 2, wherein the antibody exhibits effector function.

9. An antibody that specifically binds to human TIGIT, wherein the antibody is a human, monoclonal, full-length IgG1 subclass antibody comprising a human IgG1 Fc region, a heavy chain variable region (VH) as shown in the amino acid sequence of SEQ ID NO: 34, and a light chain variable region (VL) as shown in the amino acid sequence of SEQ ID NO:

36.

10. The antibody of claim 9, wherein the antibody exhibits effector function.

11. A polynucleotide encoding the antibody of claim 1 or 9.

12. A vector comprising the polynucleotide of claim 11.

13. A host cell comprising the vector of claim 12.

14. The host cell of claim 13, wherein the host cell is prokaryotic.

15. The host cell of claim 14, wherein the host cell is an Escherichia coli host cell.

16. The host cell of claim 13, wherein the host cell is eukaryotic.

17. The host cell of claim 16, wherein the host cell is a 293 host cell, a CHO host cell, a yeast host cell, or a plant host cell.

18. A method of producing the antibody of claim 1 or 9, the method comprising culturing the host cell of claim 13 in a culture medium.

19. The method of claim 18, wherein the method further comprises recovering the antibody from the host cell or culture medium.

20. A composition comprising the antibody of claim 1 or 9 and a pharmaceutically acceptable carrier, excipient, or diluent.

21. The composition of claim 20, wherein the composition is a pharmaceutical composition.

22. The composition of claim 20 or 21, wherein the composition further comprises a PD-1 axis binding antagonist or an additional therapeutic agent.

23. A kit comprising the antibody of claim 1 or 9 and a package insert comprising instructions for using the antibody to treat cancer or delay cancer progression in a subject.

24. A kit comprising the antibody of claim 1 or 9 and a package insert comprising instructions for using the antibody to treat or delay progression of an immune-related disease in a subject.

25. A kit comprising the antibody of claim 1 or 9 and a package insert comprising instructions for use in enhancing, increasing, or stimulating an immune response or function in a subject.

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