Anti-IL13Ra2 antibodies and uses thereof

Abstract

Anti-IL13Ra2 antibodies and antigen-binding fragments thereof are described, as well as nucleic acids encoding the antibodies, compositions comprising the antibodies, and methods of producing the antibodies and methods of using the antibodies to treat or prevent diseases, such as cancer, inflammatory diseases, and / or autoimmune diseases.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 267,681, filed February 8, 2022, the disclosure of which is incorporated by reference in its entirety herein.

[0002] FIELD OF THEINVENTION The present invention relates to monoclonal antibodies that bind to interleukin-13 receptor subunit alpha 2 (IL13Ra2), nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. Methods of making the antibodies and methods of using the antibodies to treat diseases, including cancer, inflammatory diseases, autoimmune diseases, and / or related complications are also provided.

[0003] Reference to an Electronically Submitted Sequence Listing This application contains a Sequence Listing that has been submitted electronically. The contents of the electronic Sequence Listing (065799.36WO1 Sequence Listing.xml; size: 182,964 bytes; and creation date Jan. 23, 2023) are hereby incorporated by reference in their entirety. [Background technology]

[0004] Interleukin-13 receptor alpha 2 (also known as IL13Ra2, IL-13Ra2, IL-13 receptor subunit alpha 2, IL13Rα2, CT19, IL-13R2, IL13BP and CD213A2) is a 42 kD cell surface receptor for IL-13, a proinflammatory cytokine that regulates both hematopoietic and non-hematopoietic cells (Marone, G., Granata, F. et al., Frontiers in Pharmacology (2019) 10, 1387). In addition to IL13Ra2, IL-13 also binds to IL13Ra1. IL-13 binds to IL13Ra1 with low affinity and activates the JAK-STAT signaling pathway that mediates IL-13-mediated cellular responses. Because IL13Ra2 lacks canonical JAK-STAT signaling function and IL13 binds to IL13Ra2 with high affinity, it has been proposed that IL13Ra2 functions as a decoy receptor that negatively regulates IL-13 responses (Rahaman, SO, Sharma, P., et al., Cancer Research (2002) 63, 1103-1109). More recently, it has been reported that IL13Ra2 activates AP-1-mediated transcription in certain cells (Fujisawa, T., Joshi, BH, et al., International Journal of Cancer (2012) 131, 344-356), suggesting that the function of IL13Ra2 is more complex than previously recognized. Summary of the Invention [Problem to be solved by the invention]

[0005] IL13Ra2 is overexpressed in various types of cancer cells, such as glioblastoma multiforme (GBM), pancreatic cancer, renal cell carcinoma, head and neck cancer, mesothelioma, melanoma, ovarian cancer, breast cancer, gastric cancer, and colorectal cancer (Suzuki, A., Leland, P., et al., Cytokine (2015) 75, 79-88). IL13Ra2 is expressed at low levels on limited tissue cells, making it a useful cancer biomarker and a promising therapeutic target. In addition, IL13 and IL13Ra2 are involved in promoting the proliferation and invasion of epithelial cancers (Fujisawa, T., Joshi, BH, et al., International Journal of Cancer (2012) 131, 344-356). In summary, IL13Ra2 is a tumor-associated antigen (TAA) and may be an excellent drug target. [Means for solving the problem]

[0006] In one general aspect, the invention relates to an isolated monoclonal antibody, or antigen-binding fragment thereof, that specifically binds to IL13Ra2.

[0007] (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. The present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof, which specifically binds to IL13Ra2, preferably human IL13Ra2, and comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0008] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, or 169, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170.

[0009] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; or (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170. Includes.

[0010] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof binds to IL13Ra2 and can induce effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or can mediate the recruitment of a conjugated drug, and / or can form a bispecific antibody with another monoclonal antibody or antigen-binding fragment thereof that has cancer-killing activity.

[0011] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is chimeric.

[0012] In certain embodiments, the isolated monoclonal antibody or antigen-binding fragment thereof is human or humanized. In certain embodiments, a humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212; or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

[0013] In certain embodiments, the humanized monoclonal antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213. Includes.

[0014] Also provided is an isolated bispecific antibody comprising a monoclonal antibody of the invention, or an antigen-binding fragment thereof.

[0015] Also provided is an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody of the invention.

[0016] Also provided is a vector comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention.

[0017] A host cell comprising a vector containing an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention is also provided.

[0018] In certain embodiments, a pharmaceutical composition is provided comprising an isolated monoclonal antibody or antigen-binding fragment thereof, or an isolated bispecific antibody or antigen-binding fragment thereof of the invention and a pharma- ceutically acceptable carrier.

[0019] Also provided is a method of specifically targeting IL13Ra2 on the surface of cancer cells in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the invention.

[0020] Also provided is a method for treating cancer in a subject in need thereof, comprising administering the pharmaceutical composition of the present invention to the subject.Cancer can be any liquid or solid cancer, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0021] Also provided is a method of treating an inflammatory disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the present invention.

[0022] Also provided is a method of treating an autoimmune disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the present invention.

[0023] Also provided is a method of producing a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising culturing a cell containing a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, under conditions for production of the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, and recovering the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof from the cell or culture.

[0024] Also provided is a method of producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof, or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising combining the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof with a pharma- ceutically acceptable carrier to obtain a pharmaceutical composition.

[0025] Also provided is a method for determining the level of IL13Ra2 in a subject. The method includes (a) obtaining a sample from a subject, (b) contacting the sample with an antibody or an antigen-binding fragment thereof of the present invention, and (c) determining the level of IL13Ra2 in the subject. In certain embodiments, the sample is a tissue sample. The tissue sample can be, for example, a cancer tissue sample. In certain embodiments, the sample is a blood sample.

[0026] In another general aspect, the invention relates to a chimeric antigen receptor (CAR) construct for inducing T cell-mediated cancer killing, the CAR construct comprising at least one antigen-binding domain that specifically binds human IL13Ra2, a hinge region, a transmembrane region, and an intracellular signaling domain.

[0027] Provided is an isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR). The CAR can comprise (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to IL13Ra2, (b) a hinge region, (c) a transmembrane region, and (d) an intracellular signaling domain.

[0028] In certain embodiments, the antigen binding domain comprises: (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having a polypeptide sequence of:

[0029] In certain embodiments, the antigen-binding domain comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, or 169, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170.

[0030] In certain embodiments, the antigen binding domain comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; or (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170. Includes.

[0031] In certain embodiments, the antigen-binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212; or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

[0032] In certain embodiments, the antigen-binding domain is humanized, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213. Includes.

[0033] In certain embodiments, the antigen-binding domain is a single chain variable fragment (scFv). In certain embodiments, the antigen-binding domain is a humanized single chain variable fragment (scFv).

[0034] In certain embodiments, a chimeric antigen receptor (CAR) comprises one or more antigen-binding domains.

[0035] In certain embodiments, the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0036] Also provided is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of the present invention.

[0037] Also provided is a vector comprising an isolated polynucleotide comprising a nucleic acid encoding a CAR of the invention.

[0038] Host cells comprising the vectors of the invention are also provided.

[0039] In certain embodiments, the host cell is a T cell, preferably a human T cell. In certain embodiments, the host cell is a NK cell, preferably a human NK cell. The T cell or NK cell can be engineered to express the CAR of the present invention to treat a disease, such as, for example, cancer.

[0040] Also provided is a method of making a host cell expressing a chimeric antigen receptor (CAR) of the invention, comprising transducing a T cell or an NK cell with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0041] Also provided is a method of producing a CAR-T cell or a CAR-NK cell of the invention, comprising culturing a T cell or an NK cell comprising an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) of the invention under CAR-T cell or CAR-NK cell producing conditions, and recovering the CAR-T cell or CAR-NK cell.

[0042] Also provided is a method of generating a population of RNA engineered cells comprising a chimeric antigen receptor (CAR) of the invention, the method comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) of the invention, the isolated polynucleotide being an in vitro transcribed RNA or a synthetic RNA.

[0043] Also provided are methods of treating cancer in a subject in need thereof, comprising administering to the subject the CAR-T cells and / or CAR-NK cells of the invention. The cancer can be any liquid or solid cancer, including, but not limited to, lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma, and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML), and other liquid tumors.

[0044] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that increases the efficacy of a cell expressing a CAR molecule.

[0045] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that ameliorates one or more side effects associated with administration of a cell expressing a CAR molecule.

[0046] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that treats an IL13Ra2-associated disease.

[0047] The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It is to be understood, however, that the application is not limited to the precise embodiments shown in the drawings. [Brief description of the drawings]

[0048] [Figure 1A]Figure 1 shows the binding of purified chimeric anti-IL13Ra2 mAb (VH and VL regions of mouse mAb fused to human IgG1 heavy and kappa light chain constant regions, respectively) to immobilized recombinant IL13Ra2 in an ELISA assay. The antigen was human IL13Ra2 (ACROBiosystems; Newark, DE; CAT#: IL2-H52H5). L3A6-C refers to the chimeric version of L3A6, another antibody following the same naming convention; an isotype control (IgG1) was included as a negative control antibody. [Figure 1B] This is a continuation of Figure 1A. [Figure 1C] This is a continuation of Figure 1B. [Figure 1D] This is a continuation of Figure 1C. [Figure 1E] This is a continuation of Figure 1D. [Figure 2A] Figure 1 shows binding of anti-IL13Ra2 mAb to A375 cells (ATCC; Manassas, VA; Cat#: CRL-1619) using FACS analysis. L3A6-C refers to the chimeric version of L3A6, other antibodies following the same naming convention; Isotype control (IgG1), negative control antibody. [Figure 2B] This is a continuation of Figure 2A. [Figure 3A] Figure 1 shows binding of humanized anti-IL13Ra2 mAbs to immobilized recombinant IL13Ra2 in an ELISA assay. The antigen was human IL13Ra2 (ACROBiosystems; Newark, DE; CAT#: IL2-H52H5). L3A6-H1L1 refers to a humanized mAb constructed with the L3A6-H1 heavy chain and L3A6-L1 light chain shown in Table 7; other humanized clones follow the same naming convention. An isotype control (IgG1) was included as a negative control antibody. [Figure 3B] This is a continuation of Figure 3A. [Figure 3C] This is a continuation of Figure 3B. [Figure 3D] This is a continuation of Figure 3C. [Figure 3E] This is a continuation of Figure 3D. [Figure 3F] Continuation of Figure 3E. [Figure 3G] Continuation of Figure 3F. [Figure 3H] Continuation of Figure 3G. [Figure 4A] Figure 1 shows binding of humanized anti-IL13Ra2 mAbs to A375 cells (ATCC; Manassas, VA; Cat#: CRL-1619) using FACS analysis. L3A6-H1L1 refers to a humanized mAb constructed with the L3A6-H1 heavy chain and L3A6-L1 light chain shown in Table 7; other humanized clones follow the same naming convention. Isotype control (IgG1), negative control antibody. [Figure 4B] This is a continuation of Figure 4A. [Figure 4C] This is a continuation of Figure 4B. [Figure 4D] Continuation of Figure 4C. [Figure 4E] This is a continuation of Figure 4D. [Figure 4F] Continuation of Figure 4E. [Figure 4G] Continuation of Figure 4F. [Figure 4H] Continuation of Figure 4G. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0049] Throughout the background and specification, various publications, articles and patents are cited or described. Each of these references is incorporated herein by reference in its entirety. The discussion of documents, acts, materials, devices, articles and the like which has been included in the present specification is for the purpose of providing a context for the present invention. Such discussion is not an admission that any or all of those matters form part of the prior art with respect to any invention(s) disclosed or claimed.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Otherwise, certain terms used herein have the meaning given in the specification.

[0051] It must be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0052] Unless otherwise specified, any numerical values, e.g., concentrations or concentration ranges, described herein should be understood to be modified in all cases by the term "about." Thus, numerical values ​​generally include ±10% of the recited value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, unless the context clearly dictates otherwise, the use of numerical ranges expressly includes all individual numerical values ​​within that range, including all possible subranges, integers and fractions of values ​​within such ranges.

[0053] Unless otherwise indicated, the term "at least" before a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0054] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," or "containing," or any other variation thereof, are understood to imply that the specified integer or group of integers is inclusive, but not the exclusion of any other integer or group of integers, and are non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or device containing a list of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or device. Additionally, unless expressly stated otherwise, "or" refers to an inclusive or and not an exclusive or. For example, condition A or B is satisfied by any one of the following: A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and both A and B are true (or exist).

[0055] As used herein, the conjunction "and / or" between multiple listed elements is understood to encompass the individual and combined options. For example, when two elements are joined by "and / or," the first option refers to the application of the first element without the second element. The second option refers to the application of the second element without the first element. The third option refers to the application of the first and second elements together. Any one of these options is understood to fall within this meaning and thus meet the requirements of the term "and / or" as used herein. The simultaneous application of more than one of these options is also understood to fall within this meaning and thus meet the requirements of the term "and / or."

[0056] As used herein, the term "consists of" or variations such as "consist of" or "consisting of," as used throughout the specification and claims, indicates that any recited integer or group of integers is inclusive, but that no additional integers or groups of integers can be added to the specified method, structure, or composition.

[0057] As used herein, the term "consists essentially of" or variations such as "consist essentially of" or "consisting essentially of," as used throughout the specification and claims, indicates that any recited integer or group of integers is inclusive, and, where appropriate, any recited integer or group of integers that do not materially alter the basic or novel characteristics of the specified method, structure, or composition. See MPEP § 2111.03.

[0058] As used herein, "subject" refers to any animal, preferably a mammal, most preferably a human. As used herein, the term "mammal" encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more preferably humans.

[0059] The words "right", "left", "lower" and "top" designate directions in the drawings to which reference is made.

[0060] It should also be understood that the terms "about," "approximately," "generally," "substantially," and similar terms used herein when referring to dimensions or features of preferred inventive components indicate that the described dimensions / features are not precise boundaries or parameters, as would be understood by one of ordinary skill in the art, and do not exclude minor variations therefrom that are functionally the same or similar. At a minimum, such references involving numerical parameters are expected to include variations that are expected not to alter the lowest numerical value using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.).

[0061] The terms "identical" or percent "identity," in the context of two or more nucleic acid or polypeptide sequences (e.g., anti-IL13Ra2 antibodies and the polynucleotides encoding them, chimeric antigen receptors (CARs) comprising an antigen-binding domain specific for IL13Ra2 and the polynucleotides encoding them), refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when aligned compared to maximum correspondence as determined using one of the following sequence comparison algorithms or by visual inspection.

[0062] For sequence comparison, typically one sequence serves as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the designated program parameters.

[0063] Optimal alignment of sequences for comparison can be determined, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 1981; 2:482, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 1970; 48:443, by the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 1988; 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (generally as described in Current Protocols in Molecular Biology, F. M. Ausubel et al., eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., 1995 Supplement). (see Ausubel) can be implemented.

[0064] Examples of algorithms suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., J. Mol. Biol. 1990; 215: 403-410 and Altschul et al., Nucleic Acids Res. 1997; 25: 3389-3402, respectively. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that match or meet some positive threshold score T when aligned with words of the same length in the database sequences. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased.

[0065] Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matched residues; always >0) and N (penalty score for mismatched residues; always <0). For amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. Extension of the word hits in each direction is stopped if: the cumulative alignment score falls an amount X from its maximum achieved value; the cumulative score falls below zero due to the accumulation of one or more negative scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 1989;89:10915).

[0066] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 1993; 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the minimum sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered to be similar to a reference sequence if the minimum sum probability in the comparison between the test nucleic acid and the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0067] A further indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, for example, where the two peptides differ only by conservative substitutions, a polypeptide will generally be substantially identical to a second polypeptide. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions.

[0068] As used herein, the term "isolated" means that a biological component (e.g., a nucleic acid, peptide, or protein) has been substantially separated, produced separately, or purified from other biological components of the organism in which it naturally occurs, i.e., from other chromosomal and extrachromosomal DNA and RNA, and proteins. Thus, "isolated" nucleic acids, peptides, and proteins include nucleic acids and proteins purified by standard purification methods. "Isolated" nucleic acids, peptides, and proteins can be part of a composition and still be isolated, if the composition is not part of the natural environment of the nucleic acid, peptide, or protein. The term also encompasses nucleic acids, peptides, and proteins prepared by recombinant expression in a host cell, as well as chemically synthesized nucleic acids.

[0069] As used herein, the term "polynucleotide", which is interchangeably referred to as "nucleic acid molecule", "nucleotide" or "nucleic acid", refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. "Polynucleotide" includes, but is not limited to, single-stranded and double-stranded DNA, DNA that is a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA that is a mixture of single-stranded and double-stranded regions, hybrid molecules containing DNA and RNA that may be single-stranded or, more commonly, double-stranded, or a mixture of single-stranded and double-stranded regions. Furthermore, "polynucleotide" refers to triple-stranded regions that include RNA or DNA, or both RNA and DNA. The term polynucleotide also includes DNA or RNA that contain one or more modified bases, and DNA or RNA with backbones modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and unusual bases, such as inosine. A variety of modifications can be made to DNA and RNA. Thus, "polynucleotide" includes chemically, enzymatically, or metabolically modified forms of polynucleotides commonly found in nature, as well as the DNA and RNA chemical forms characteristic of viruses and cells. "Polynucleotide" also includes relatively short nucleic acid strands often referred to as oligonucleotides.

[0070] As used herein, the term "vector" is a replicon into which another nucleic acid segment may be operatively inserted so as to bring about the replication or expression of the segment.

[0071] As used herein, the term "host cell" refers to a cell that contains a nucleic acid molecule of the invention. A "host cell" can be any cell type, e.g., a primary cell, a cultured cell, or a cell from a cell line. In one embodiment, a "host cell" is a cell that has been transfected with a nucleic acid molecule of the invention. In another embodiment, a "host cell" is the progeny or potential progeny of such a transfected cell. The progeny of a cell may or may not be identical to the parent cell, e.g., due to mutations or environmental influences that may occur in subsequent generations, or due to integration of the nucleic acid molecule into the host cell genome.

[0072] The term "expression" as used herein refers to the biosynthesis of a gene product. The term includes the transcription of a gene into RNA. The term also includes the translation of RNA into one or more polypeptides, and further includes all naturally occurring post-transcriptional and post-translational modifications. The expressed antibody may be in the cytoplasm of the host cell, in the extracellular environment, for example in the growth medium of a cell culture, or may be anchored to the cell membrane.

[0073] As used herein, the term "peptide", "polypeptide", or "protein" can refer to a molecule composed of amino acids and can be recognized as a protein by those skilled in the art. Conventional one-letter or three-letter codes for amino acid residues are used herein. The terms "peptide", "polypeptide", and "protein" can be used interchangeably herein to refer to a polymer of amino acids of any length. The polymer can be linear or branched, can contain modified amino acids, and can be interrupted by non-amino acids. The term also encompasses amino acid polymers that are modified naturally or by intervention, such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are, for example, polypeptides containing one or more analogs of amino acids (including, for example, unnatural amino acids, etc.), and other modifications known in the art.

[0074] The peptide sequences described herein are written according to the usual convention whereby the N-terminal region of the peptide is on the left and the C-terminal region is on the right Although isomeric forms of amino acids are known, unless expressly indicated otherwise, the amino acids represented are in the L-form.

[0075] Chimeric antigen receptors (CARs) As used herein, the term "chimeric antigen receptor" (CAR) refers to a recombinant polypeptide that includes at least an extracellular domain that specifically binds an antigen or target, a transmembrane domain, and an intracellular T cell receptor activation signaling domain. When the extracellular domain of the CAR binds to a target antigen on the surface of a target cell, clustering of the CAR occurs, providing an activation stimulus to the cell containing the CAR. The CAR redirects the specificity of immune effector cells, triggering proliferation, cytokine production, phagocytosis, and / or production of molecules that can mediate cell death of target antigen-expressing cells in a major histocompatibility (MHC)-independent manner.

[0076] In one embodiment, the CAR comprises an antigen-binding domain, a hinge region, a costimulatory domain, an activation domain, and a transmembrane region. In one embodiment, the CAR comprises an antigen-binding domain, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region. In one embodiment, the CAR comprises two antigen-binding domains, a hinge region, a costimulatory domain, an activation domain, and a transmembrane region. In one embodiment, the CAR comprises two antigen-binding domains, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region.

[0077] As used herein, the term "signal peptide" refers to a leader sequence at the amino-terminus (N-terminus) of the nascent CAR protein that co- or post-translationally targets the nascent protein to the endoplasmic reticulum, leading to subsequent surface expression.

[0078] As used herein, the term "extracellular antigen-binding domain," "extracellular domain," or "extracellular ligand-binding domain" refers to the portion of a CAR that is located on the outside of the cell membrane and can bind to an antigen, target, or ligand.

[0079] As used herein, the term "hinge region" refers to the portion of a CAR that connects two adjacent domains of a CAR protein, e.g., the extracellular domain and the transmembrane domain.

[0080] As used herein, the term "transmembrane domain" refers to the portion of the CAR that extends across the cell membrane and anchors the CAR to the cell membrane. This may be referred to as the "transmembrane region."

[0081] Costimulatory domain As used herein, chimeric antigen receptors can incorporate costimulatory (signaling) domains to enhance their efficacy. The costimulatory (signaling) domains can be derived from costimulatory molecules, which are cell surface molecules other than the antigen receptor or its ligand that are required for an efficient immune response.The costimulatory domain may be derived from costimulatory molecules, including CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death-1 (PD-1), inducible T cell costimulatory factor (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Ig alpha (CD79a), DAP10, Fc gamma receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD8 alpha, CD8 beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGAE, CD103, ITGAL, CD1a, CD1b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITGB 2(CD18), ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRTAM, Ly9(CD229), CD 160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor, or any combination thereof.

[0082] Activation domain As used herein, chimeric antigen receptor can include an activation domain.Activation domain can include, but is not limited to, CD3.CD3 is a component of the T cell receptor on natural T cells and has been shown to be an important intracellular activation component in CAR.In a preferred embodiment, CD3 is CD3 zeta.

[0083] Hinge Area As described herein, the chimeric antigen receptor can include a hinge region, which is part of the extracellular domain and is sometimes referred to as the "spacer" region. A variety of hinges can be used in accordance with the present invention, including the costimulatory molecules described above, immunoglobulin (Ig) sequences, or other appropriate molecules to achieve the desired specific distance from the target cell. In some embodiments, the entire extracellular domain comprises the hinge region.

[0084] transmembrane region As used herein, a chimeric antigen receptor (CAR) can include a transmembrane region / domain. A CAR can be designed to include a transmembrane domain fused to the extracellular domain of the CAR. Similarly, it can be fused to the intracellular domain of the CAR. In one embodiment, a transmembrane domain that is naturally associated with one of the domains in the CAR is used. In some cases, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex. The transmembrane domain can be derived from either natural or synthetic sources. If the source is natural, the domain can be derived from any membrane-bound or transmembrane protein. Transmembrane regions that are particularly useful in the present invention include CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death-1 (PD-1), inducible T cell costimulator (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14;TNFSF14), NKG2C, Ig alpha (CD79a), DAP10, Fc gamma receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD8 alpha, CD8 beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGAE, CD103, ITGAL, CD1a, CD1b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITGB 2(CD18), ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRTAM, Ly9(CD229), CD 160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Lyl08), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor, immunoglobulin protein, or a fragment or any combination thereof;

[0085] immune cells In certain aspects, the present invention provides a cell that is an immune cell comprising an isolated polynucleotide comprising a nucleotide sequence encoding a CAR as provided herein or a vector comprising an isolated polynucleotide. An immune cell comprising an isolated polynucleotide and / or vector of the present invention can be referred to as an "engineered immune cell." Preferably, the engineered immune cell is of human origin (is a cell of human origin prior to being recombined).

[0086] The engineered immune cells can be, for example, of lymphoid lineage. Non-limiting examples of lymphoid lineage include T cells and natural killer (NK) cells. T cells express the T cell receptor (TCR), with most cells expressing α and β chains, and a smaller population expressing γ and δ chains. T cells useful as engineered immune cells of the present invention include CD4 + or CD8 + and T helper cells (CD4 + ), cytotoxic T cells (also called cytotoxic T lymphocytes, CTLs; CD8 + Examples of immune cells include, but are not limited to, immune cells (e.g., immune cells), and memory T cells, including central memory T cells, stem-like memory T cells, and effector memory T cells, natural killer T cells, mucosal-associated invariant T cells, and γδ T cells. Other exemplary immune cells include, but are not limited to, macrophages, antigen-presenting cells (APCs), or any immune cell that expresses an inhibitor of a cell-mediated immune response, such as an immune checkpoint inhibitor pathway receptor (e.g., PD-1). Precursor cells of immune cells that can be used according to the present invention include hematopoietic stem and / or progenitor cells. Hematopoietic stem and / or progenitor cells can be derived from bone marrow, umbilical cord blood, adult peripheral blood after cytokine mobilization, etc., by methods known in the art. The immune cells are engineered to recombinantly express the CAR of the present invention.

[0087] Immune cells and their precursor cells can be isolated by methods known in the art, including commercially available methods (see, for example, Rowland Jones et al., Lymphocytes: A Practical Approach, Oxford University Press, NY 1999). Sources of immune cells or their precursors include, but are not limited to, peripheral blood, umbilical cord blood, bone marrow, or other sources of hematopoietic cells. Various techniques can be used to separate cells to isolate or enrich for desired immune cells. For example, negative selection methods can be used to remove cells that are not the desired immune cells. Additionally, positive selection methods can be used to isolate or enrich for desired immune cells or their precursors, or positive and negative selection methods can be used in combination. When isolating a particular type of cell, such as a particular T cell, various cell surface markers or combinations of markers (e.g., CD3, CD4, CD8, CD34) can be used to separate the cells.

[0088] The immune cells or their precursor cells can be autologous (autologous) or non-autologous (non-autologous) to the subject to whom they are administered in the treatment method of the present invention. Autologous cells are isolated from the subject to whom the engineered immune cells recombinantly expressing the CAR are administered. If desired, the cells can be obtained by leukapheresis, in which leukocytes are selectively removed from drawn blood, recombined, and then reinfused back into the donor. Alternatively, allogeneic cells from a non-autologous donor who is not the subject can be used. In the case of a non-autologous donor, the cells are sorted and matched with human leukocyte antigens (HLA) to determine the appropriate level of compatibility. For both autologous and non-autologous cells, the cells can be cryopreserved, if desired, until ready for use.

[0089] Various methods of isolating immune cells that can be used for recombinant expression of the CAR of the invention have been described and can be used, including using peripheral blood donor lymphocytes (Sadelain et al., Nat. Rev. Cancer 2003; 3:35-45; Morgan et al., Science 2006; 314:126-9), using lymphocyte cultures derived from tumor infiltrating lymphocytes (TILs) in tumor biopsies (Panelli et al., J. Immunol. 2000; 164:495-504; Panelli et al., J. Immunol. 2000; 164:4382-92), and using antigen-specific peripheral blood leukocytes selectively expanded in vitro using artificial antigen presenting cells (AAPCs) or dendritic cells (Dupont et al., Cancer Res. 2005; 65:5417-427; Papanicolaou et al., Blood 2003; 102:2498-505). When using stem cells, the cells can be isolated by methods well known in the art (e.g., see Klug et al., Hematopoietic Stem Cell Protocols, Humana Press, NJ 2002; Freshney et al., Culture of Human Stem Cells, John Wiley & Sons 2007).

[0090] According to certain embodiments, the method of producing engineered immune cells comprises transfecting or transducing immune effector cells isolated from an individual to express one or more CARs according to embodiments of the present invention. Methods of preparing immune cells for immunotherapy are described, for example, in WO2014 / 130635, WO2013 / 176916 and WO2013 / 176915, which are incorporated herein by reference. Individual steps that can be used for the preparation of engineered immune cells are disclosed, for example, in WO2014 / 039523, WO2014 / 184741, WO2014 / 191128, WO2014 / 184744 and WO2014 / 184143, which are incorporated herein by reference.

[0091] In one specific embodiment, immune effector cells, such as T cells, are genetically modified with a CAR of the invention (e.g., transduced with a viral vector comprising a nucleic acid encoding the CAR) and then activated and expanded in vitro. In various embodiments, T cells can be activated and expanded before or after genetic modification to express a CAR using methods described, for example, in US6,352,694, US6,534,055, US6,905,680, US6,692,964, US5,858,358, US6,887,466, US6,905,681, US7,144,575, US7,067,318, US7,172,869, US7,232,566, US7,175,843, US5,883,223, US6,905,874, US6,797,514, US6,867,041, US2006 / 121005, which are incorporated herein by reference. T cells can be expanded in vitro or in vivo. In general, the T cells of the present invention can be expanded by contact with a surface having attached thereto an agent that stimulates CD3 / TCR complex-associated signals and a ligand that stimulates costimulatory molecules on the surface of the T cells. As a non-limiting example, a population of T cells can be stimulated, for example, by contact with an anti-CD3 antibody, or an antigen-binding fragment thereof, or an anti-CD3 antibody immobilized on the surface, or by contact with a protein kinase C activator (e.g., bryostatin) in combination with a calcium ionophore, or by activation of the CAR itself, as described herein. To costimulate accessory molecules on the surface of the T cells, a ligand that binds to the accessory molecule is used. For example, a population of T cells can be contacted with an anti-CD3 antibody and an anti-CD28 antibody under conditions appropriate to stimulate proliferation of the T cells.Suitable conditions for T cell culture include, for example, a suitable medium (e.g., Minimum Essential Medium or RPMI Media 1640 or X-vivo 5 (Lonza)) that may contain factors necessary for growth and survival, including serum (e.g., fetal bovine serum or human serum), cytokines such as IL-2, IL-7, IL-15, and / or IL-21, insulin, IFN-g, GM-CSF, TGFβ, and / or other additives for cell growth known to those of skill in the art. In other embodiments, T cells can be activated and stimulated with feeder cells and suitable antibodies and cytokines to grow using methods such as those described in US6,040,177, US5,827,642, and WO2012129514, which are incorporated herein by reference.

[0092] Antibodies and antigen-binding domains The present invention generally relates to isolated anti-IL13Ra2 antibodies, chimeric antigen receptors (CARs), nucleic acids and expression vectors encoding the antibodies and CARs, recombinant cells containing the vectors, and compositions comprising the antibodies, CARs, and recombinant cells expressing the CARs. The present invention also generally relates to methods of making the antibodies and CARs, and methods of using the antibodies and CARs to treat diseases, which may include, but are not limited to, cancer, inflammatory diseases, and autoimmune diseases. The antigen-binding domains of the antibodies and CARs of the present invention have one or more desirable functional properties, including, but not limited to, high affinity binding to IL13Ra2, high specificity for IL13Ra2, the ability to stimulate complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADCP), and / or antibody-dependent cellular cytotoxicity (ADCC) against cells expressing IL13Ra2, and the ability to inhibit tumor growth in subjects and animal models when administered alone or in combination with other anti-cancer therapies.

[0093] In a general aspect, the invention relates to an isolated monoclonal antibody, or antigen-binding fragment thereof, that binds to IL13Ra2.

[0094] As used herein, the term "antibody" is used in a broad sense and includes immunoglobulin or antibody molecules, including human antibodies, humanized antibodies, composite antibodies, and chimeric antibodies, as well as antibody fragments that are monoclonal or polyclonal. In general, an antibody is a protein or peptide chain that exhibits binding specificity to a specific antigen. Antibody structure is well known. Immunoglobulins can be assigned to five major classes (i.e., IgA, IgD, IgE, IgG, and IgM) depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. Thus, the antibodies of the present invention can be of any of the five major classes or corresponding subclasses. Preferably, the antibodies of the present invention are IgG1, IgG2, IgG3, or IgG4. Antibody light chains of vertebrate species can be assigned to one of two clearly distinct types, namely kappa and lambda, based on the amino acid sequence of their constant domains. Thus, the antibodies of the present invention can contain a kappa or lambda light chain constant domain. According to certain embodiments, the antibodies of the invention comprise the heavy and / or light chain constant regions of a rat or human antibody. In addition to the heavy and light chain constant domains, the antibodies contain an antigen-binding region composed of a light chain variable region and a heavy chain variable region, each of which contains three domains (i.e., complementarity determining regions 1-3; CDR1, CDR2 and CDR3). The light chain variable region domains are alternatively referred to as LCDR1, LCDR2 and LCDR3, and the heavy chain variable region domains are alternatively referred to as HCDR1, HCDR2 and HCDR3.

[0095] "Complementarity determining regions" (CDRs) are antibody regions that bind antigens. CDRs can be defined using various descriptions, for example, Kabat (Wu et al., J Exp Med 132: 211-50, 1970) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), Chothia (Chothia et al., J Mol Biol 196: 901-17, 1987), IMGT (Lefranc et al., Dev Comp Immunol 27: 55-77, 2003) and AbM (Martin and Thornton J Bmol Biol 263: 800-15, 1996). Correspondence between various outlines and variable region numbering has been described (see, e.g., Lefranc et al., Dev Comp Immunol 27: 55-77, 2003; Honegger and Pluckthun, J Mol Biol 309:657-70, 2001; International ImmunoGeneTics (IMGT) database, web resource, http: / / www_imgt_org). CDRs can be delineated using available programs, e.g., abYsis by UCL Business PLC. The terms "CDR", "HCDR1", "HCDR2", "HCDR3", "LCDR1", "LCDR2" and "LCDR3", as used herein, include CDRs as defined by any of the above methods, Kabat, Chothia, IMGT or AbM, unless expressly stated otherwise herein. For example, the correspondence between numbering systems, including the Kabat numbering and the IMGT specific numbering system, is well known to those of skill in the art (see, for example, Kabat, Chothia, Martin, Lefranc et al.).

[0096] [Table 1-1]

[0097] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to IL13Ra2 is substantially free of antibodies that do not bind to the same IL13Ra2). Moreover, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0098] As used herein, the term "monoclonal antibody" 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 that may be present in minor amounts. The monoclonal antibodies of the invention can be produced by hybridoma methods, phage display techniques, single lymphocyte gene cloning techniques, or by recombinant DNA methods. For example, monoclonal antibodies can be produced by hybridomas comprising B cells obtained from a transgenic non-human animal, e.g., a transgenic mouse or rat, whose genome comprises a human heavy chain transgene and a light chain transgene.

[0099] As used herein, the terms "antigen-binding fragment" and / or "antigen-binding domain" refer to antibody fragments such as, for example, diabodies, Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabodies (ds diabodies), single-chain antibody molecules (scFv), single domain antibodies (sdab), scFv dimers (bivalent diabodies), multispecific antibodies formed from a portion of an antibody comprising one or more CDRs, camelized single domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment that binds to an antigen but does not comprise a complete antibody structure. An antigen-binding fragment can bind to the same antigen as the parent antibody or parent antibody fragment binds. According to certain embodiments, the antigen-binding fragment comprises a light chain variable region, a light chain constant region, and an Fd segment of a heavy chain. According to other particular embodiments, the antigen-binding fragment comprises Fab and F(ab'). The antigen-binding domain is capable of binding to the same antigen as the parent antibody binds. According to particular embodiments, the antigen-binding domain comprises a single chain antibody molecule (scFv).

[0100] As used herein, the term "single chain antibody" refers to a conventional single chain antibody in the art that comprises a heavy chain variable region and a light chain variable region linked by a short peptide of about 15 to about 20 amino acids. As used herein, the term "single domain antibody" refers to a conventional single domain antibody in the art that comprises a heavy chain variable region and a heavy chain constant region or comprises only a heavy chain variable region.

[0101] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody having an amino acid sequence corresponding to an antibody produced by a human made using any technique known in the art. This definition of a human antibody includes intact or full-length antibodies, fragments thereof, and / or antibodies that comprise at least one human heavy and / or light chain polypeptide.

[0102] As used herein, the terms "humanized antibody" and / or "humanized antigen-binding domain" refer to a non-human antibody and / or a non-human antigen-binding domain that has been modified to increase sequence homology with a human antibody and / or human antigen-binding domain, such that the antibody and / or antigen-binding domain retains its antigen-binding properties but reduces its antigenicity in the human body.

[0103] As used herein, the terms "chimeric antibody" and / or "chimeric antigen-binding domain" refer to antibodies and / or antigen-binding domains in which the amino acid sequences of the immunoglobulin molecule are derived from two or more species. The variable regions of both the light and heavy chains often correspond to the variable regions of antibodies and / or antigen-binding domains derived from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity and capacity, while the constant regions correspond to the sequences of antibodies and / or antigen-binding domains derived from another species of mammal (e.g., human) to avoid eliciting an immune response in that species.

[0104] As used herein, the term "multispecific antibody" refers to an antibody comprising a plurality of immunoglobulin variable domain sequences, where a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., on the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes do not overlap or substantially do not overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., on different proteins (or different subunits of a multimeric protein). In one embodiment, the multispecific antibody comprises a third, fourth, or fifth immunoglobulin variable domain. In one embodiment, the multispecific antibody is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

[0105] As used herein, the term "bispecific antibody" refers to a multispecific antibody that binds up to two epitopes or two antigens. A bispecific antibody is characterized by a first immunoglobulin variable domain sequence that has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In one embodiment, the first and second epitopes are on the same antigen, e.g., on the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes are on different antigens, e.g., on different proteins (or different subunits of a multimeric protein). In one embodiment, a bispecific antibody comprises a heavy chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a first epitope, and a heavy chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a second epitope. In one embodiment, the bispecific antibody comprises a half antibody or fragment thereof having binding specificity for a first epitope and a half antibody or fragment thereof having binding specificity for a second epitope. In one embodiment, the bispecific antibody comprises a scFv or fragment thereof having binding specificity for a first epitope and a scFv or fragment thereof having binding specificity for a second epitope. In one embodiment, the first epitope is located on the IL13Ra2 of the invention and the second epitope is located on PD-1, PD-L1, TIM-3, LAG-3, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, CD3, CD73, CD47, TIP-1, GPC-3, Apelin, DLL3, folate receptor alpha, Claudin18.2, and / or other tumor-associated immunosuppressive factors or surface antigens. In one embodiment, the first and second epitopes are located on the same IL13Ra2 of the invention.

[0106] As used herein, an antibody and / or antigen-binding domain that "specifically binds to IL13Ra2" has a KD of 1×10 -7 M or less, preferably 1×10 -8M or less, preferably 5×10 -9 M or less, 1×10 -9 M or less, 5×10 -10 M or less, or 1 x 10 -10 The term "KD" refers to an antibody and / or antigen-binding domain that binds to IL13Ra2, preferably human IL13Ra2, with a molecular weight of 0.1 or less. The term "KD" refers to the dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed as a molar concentration (M). The KD value for an antibody and / or antigen-binding domain can be determined in light of the present disclosure using methods in the art. For example, the KD of an antibody and / or antigen-binding domain can be determined by using surface plasmon resonance, for example by using a biosensor system, such as a Biacore® system, or by using biolayer interference technology, such as an Octet RED96 system.

[0107] The smaller the KD value of an antibody and / or antigen-binding domain, the higher the affinity with which the antibody and / or antigen-binding domain binds to a target antigen.

[0108] As used herein, the term "IC 50 " refers to the half maximal inhibitory concentration of a monoclonal or bispecific antibody or antigen-binding fragment thereof of the invention. IC 50 is a measure of the potency of a monoclonal or bispecific antibody or antigen-binding fragment thereof of the invention to inhibit binding of a target antigen (i.e., receptor or ligand) to its natural ligand or receptor, or to inhibit the function of a target antigen in a cell. In certain embodiments, a monoclonal antibody or antigen-binding fragment thereof or a bispecific antibody or antigen-binding fragment thereof has an activity of about 10 -7 Less than M, about 10 -8 Less than M, about 10 -9 Less than M, about 10 -10 Less than M, about 10 -11 Less than M, about 10 -12 Less than M or about 10 -13 Has a KD of less than M.

[0109] As used herein, "EC 50 " refers to the half maximal effective concentration of a monoclonal or bispecific antibody or antigen-binding fragment thereof of the invention. EC 50 refers to the concentration of a monoclonal or bispecific antibody or antigen-binding fragment thereof to induce a biological response (i.e., cell death) halfway between baseline and maximum over a particular exposure period. In certain embodiments, the monoclonal antibody or antigen-binding fragment thereof or the bispecific antibody or antigen-binding fragment thereof has an EC of less than about 1 mM, about 1000 nM to about 100 nM, about 100 nM to about 10 nM, about 10 nM to about 1 nM, about 1000 pM to about 500 pM, about 500 pM to about 200 pM, less than about 200 pM, about 200 pM to about 150 pM, about 200 pM to about 100 pM, about 100 pM to about 10 pM, or about 10 pM to about 1 pM. 50 has.

[0110] According to a particular aspect, the present invention provides an isolated monoclonal antibody or antigen-binding fragment thereof, or a chimeric antigen receptor (CAR) comprising an antigen-binding domain, the monoclonal antibody or antigen-binding fragment thereof, or the antigen-binding domain comprising: (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. The present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof, or chimeric antigen receptor (CAR), which specifically binds to IL13Ra2, preferably human IL13Ra2, and comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, and the antibody or antigen-binding fragment thereof or antigen-binding domain thereof has a polypeptide sequence of:

[0111] According to another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof or chimeric antigen receptor (CAR) comprising an antigen-binding domain, wherein the monoclonal antibody or antigen-binding fragment thereof or antigen-binding domain has at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, 100%, 102%, 104%, 106%, 108%, 109%, 110%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 130%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169, 169, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 179, 179, 180%, 181%, 182%, 183%, 184%, 185%, 185%, 186%, 187%, 188%, 189%, 189, 190%, 191%, 192%, 193%, 1 or 99% identical to one of SEQ ID NOs: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170, or a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, for example 95%, 96%, 97%, 98%, or 99% identical to one of SEQ ID NOs: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170, or an isolated monoclonal antibody or antigen-binding fragment thereof, or a chimeric antigen receptor (CAR).

[0112] According to another particular aspect, the present invention comprises (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; or (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170. The present invention relates to an isolated monoclonal antibody or an antigen-binding fragment or an antigen-binding domain thereof comprising:

[0113] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1, and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2.

[0114] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 17, 18, 19, 20, 21, and 22, respectively, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 16. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:15, and a light chain variable region having the polypeptide sequence of SEQ ID NO:16.

[0115] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 31, 32, 33, 34, 35, and 36, respectively, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 30. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:29, and a light chain variable region having the polypeptide sequence of SEQ ID NO:30.

[0116] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 45, 46, 47, 48, 49, and 50, respectively, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 43, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 44. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:43, and a light chain variable region having the polypeptide sequence of SEQ ID NO:44.

[0117] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 59, 60, 61, 62, 63, and 64, respectively, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 57, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 58. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:57, and a light chain variable region having the polypeptide sequence of SEQ ID NO:58.

[0118] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 73, 74, 75, 76, 77, and 78, respectively, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 71, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 72. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:71, and a light chain variable region having the polypeptide sequence of SEQ ID NO:72.

[0119] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 87, 88, 89, 90, 91, and 92, respectively, or SEQ ID NOs: 93, 94, 95, 96, 97, and 98, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 85, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 86. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:85, and a light chain variable region having the polypeptide sequence of SEQ ID NO:86.

[0120] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 101, 102, 103, 104, 105, and 106, respectively, or SEQ ID NOs: 107, 108, 109, 110, 111, and 112, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 99, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 100. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:99, and a light chain variable region having the polypeptide sequence of SEQ ID NO:100.

[0121] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 115, 116, 117, 118, 119, and 120, respectively, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 113, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 114. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:113 and a light chain variable region having the polypeptide sequence of SEQ ID NO:114.

[0122] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 129, 130, 131, 132, 133, and 134, respectively, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 127, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 128. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:127, and a light chain variable region having the polypeptide sequence of SEQ ID NO:128.

[0123] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 143, 144, 145, 146, 147, and 148, respectively, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 141, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 142. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:141, and a light chain variable region having the polypeptide sequence of SEQ ID NO:142.

[0124] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 155, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 156. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:155, and a light chain variable region having the polypeptide sequence of SEQ ID NO:156.

[0125] In one embodiment, the invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having the polypeptide sequences of SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. In another embodiment, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 169, and a light chain variable region having a polypeptide sequence that is at least 85%, preferably 90%, more preferably 95% or more, such as 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 170. Preferably, the isolated monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO:169, and a light chain variable region having the polypeptide sequence of SEQ ID NO:170.

[0126] In another particular aspect, the present invention relates to the isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, which is chimeric.

[0127] In another particular aspect, the present invention relates to an isolated monoclonal antibody or antigen-binding fragment thereof of the present invention, wherein the antibody or antigen-binding fragment thereof is human or humanized.

[0128] In another particular embodiment, the humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212; or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

[0129] In another particular embodiment, the humanized monoclonal antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213. Includes.

[0130] According to another particular embodiment, the antigen-binding domain is a single chain variable fragment (scFv) that specifically binds to IL13Ra2, preferably human IL13Ra2.

[0131] In certain embodiments, the encoded antigen-binding domain is a humanized single chain variable fragment (scFv) that specifically binds to IL13Ra2, preferably human IL13Ra2. In certain embodiments, the antigen-binding domain is a humanized single chain variable fragment (scFv) that specifically binds to IL13Ra2, preferably human IL13Ra2.

[0132] In certain embodiments, the encoded antigen-binding domain is a humanized single chain variable fragment (scFv) that specifically binds to IL13Ra2, preferably human IL13Ra2. In certain embodiments, the antigen-binding domain is a humanized single chain variable fragment (scFv) that specifically binds to IL13Ra2, preferably human IL13Ra2.

[0133] According to another specific embodiment, the chimeric antigen receptor comprises one or more antigen-binding domains.

[0134] According to another particular embodiment, the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0135] In another general aspect, the present invention relates to an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof and / or bispecific antibody or antigen-binding fragment thereof of the present invention. In another general aspect, the present invention relates to an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR), the CAR comprising the antigen-binding domain of the present invention. It will be understood by those skilled in the art that the coding sequence of a protein can be altered (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Thus, it will be understood by those skilled in the art that the nucleic acid sequence encoding the monoclonal antibody or antigen-binding fragment thereof of the present invention can be modified without changing the amino acid sequence of the protein.

[0136] In another general aspect, the present invention relates to a vector comprising an isolated nucleic acid encoding a monoclonal antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, and / or a CAR of the present invention. In light of this disclosure, any vector known to the skilled artisan can be used, for example, a plasmid, a cosmid, a phage vector, or a viral vector. In some embodiments, the vector is a recombinant expression vector, for example, a plasmid. The vector can include any element for establishing the conventional function of an expression vector, for example, a promoter, a ribosome binding element, a terminator, an enhancer, a selection marker, and an origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. Several expression vectors capable of delivering nucleic acids to cells are known in the art and can be used herein for the production of an antibody or antigen-binding fragment thereof in a cell. Conventional cloning techniques or artificial gene synthesis can be used to generate the recombinant expression vector according to embodiments of the present invention.

[0137] In another general aspect, the present invention relates to a host cell comprising an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof and / or bispecific antibody or antigen-binding fragment thereof of the present invention. In light of the present disclosure, any host cell known to the skilled artisan can be used for recombinant expression of the antibody or antigen-binding fragment thereof of the present invention. In some embodiments, the host cell is an E. coli TG1 or BL21 cell (e.g., for expression of scFv or Fab antibodies), a CHO-DG44 or CHO-K1 cell, or a HEK293 cell (e.g., for expression of full-length IgG antibodies). According to certain embodiments, the recombinant expression vector is transformed into the host cell by conventional methods, e.g., chemical transfection, heat shock, or electroporation, where it is stably integrated into the host cell genome so that the recombinant nucleic acid is effectively expressed.

[0138] In another general aspect, the invention relates to a method of producing a monoclonal antibody or antigen-binding fragment thereof and / or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof or the bispecific antibody or antigen-binding fragment thereof under conditions for production of the monoclonal antibody or antigen-binding fragment thereof or the bispecific antibody or antigen-binding fragment thereof of the invention, and recovering the antibody or antigen-binding fragment thereof from the cell or cell culture (e.g., from the supernatant). The expressed antibody or antigen-binding fragment thereof can be harvested from the cells and purified as described herein according to routine techniques known in the art.

[0139] In another general aspect, the invention relates to a cell transduced with a vector comprising an isolated nucleic acid encoding a CAR of the invention. The term "transduced" or "transduction" refers to a process in which an exogenous nucleic acid is transferred or introduced into a host cell. A "transduced" cell is a cell that has been transduced with an exogenous nucleic acid. The cell includes the primary subject cell and its progeny. In certain embodiments, the cell is a CAR-T cell, preferably a human CAR-T cell, and the T cell is engineered to express a CAR of the invention to treat a disease, e.g., cancer. In certain embodiments, the cell is a CAR-NK cell, preferably a human CAR-NK cell, and the NK cell engineered to express a CAR of the invention is used to treat a disease, e.g., cancer.

[0140] In another general aspect, the invention relates to a method of generating a CAR-T cell by transducing a T cell with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0141] In another general aspect, the invention relates to a method of producing a chimeric antigen receptor (CAR) of the invention, comprising culturing a T cell comprising a nucleic acid encoding the CAR under CAR-T cell production conditions, and recovering the CAR-T cell.

[0142] In another general aspect, the invention relates to a method of generating a CAR-NK cell by transducing an NK cell with a vector comprising an isolated nucleic acid encoding a CAR of the invention.

[0143] In another general aspect, the invention relates to a method of producing a chimeric antigen receptor (CAR) of the invention, comprising culturing an NK cell comprising a nucleic acid encoding the CAR under CAR-NK cell producing conditions, and recovering the CAR-NK cell.

[0144] In another general aspect, the invention relates to a method of generating a population of RNA engineered cells comprising a chimeric antigen receptor (CAR) of the invention. The method comprises contacting the population of cells with an isolated polynucleotide comprising a nucleic acid encoding a CAR of the invention, where the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0145] Pharmaceutical Compositions In another general aspect, the invention relates to a pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, an isolated polynucleotide, an isolated polypeptide, a host cell, and / or an engineered immune cell of the invention, and a pharma- ceutically acceptable carrier.

[0146] The term "pharmaceutical composition" as used herein refers to an article of manufacture comprising an isolated polynucleotide of the invention, an isolated polypeptide of the invention, a host cell of the invention, an engineered immune cell of the invention, an anti-IL13Ra2 monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody of the invention together with a pharma- ceutical acceptable carrier. The polynucleotides, polypeptides, host cells, engineered immune cells, anti-IL13Ra2 monoclonal antibodies or antigen-binding fragments thereof, and / or bispecific antibodies of the invention and compositions comprising them are also useful in the manufacture of medicaments for the therapeutic applications described herein.

[0147] As used herein, the term "carrier" refers to any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid-containing vesicle, microsphere, liposomal encapsulation or other material known in the art for use in pharmaceutical formulations. It is understood that the characteristics of the carrier, excipient or diluent will depend on the route of administration for a particular application. As used herein, the term "pharmaceutical acceptable carrier" refers to a non-toxic material that does not interfere with the efficacy of the composition according to the present invention or the biological activity of the composition according to the present invention. According to certain embodiments, any pharmaceutical acceptable carrier suitable for use in antibody pharmaceutical compositions in light of the present disclosure can be used in the present invention.

[0148] Formulation of pharma- ceutically active ingredients with pharma- ceutically acceptable carriers is known in the art, e.g., Remington: The Science and Practice of Pharmacy (e.g., 21st Edition (2005) and any later editions). Non-limiting examples of additional ingredients include: buffers, diluents, solvents, tonicity modifiers, preservatives, stabilizers, and chelating agents. One or more pharma-ceutically acceptable carriers can be used in formulating the pharmaceutical compositions of the present invention.

[0149] In one embodiment of the present invention, the pharmaceutical composition is a liquid formulation. A preferred example of a liquid formulation is an aqueous formulation, i.e. a formulation that contains water. The liquid formulation can include solutions, suspensions, emulsions, microemulsions, gels, etc. An aqueous formulation typically contains at least 50% w / w water, or at least 60%, 70%, 75%, 80%, 85%, 90%, or at least 95% w / w water.

[0150] In one embodiment, the pharmaceutical composition can be formulated as an injectable that can be injected, for example, via an injection device (e.g., a syringe or an infusion pump). The injection can be delivered, for example, subcutaneously, intramuscularly, intraperitoneally, intravitreally, or intravenously.

[0151] In another embodiment, the pharmaceutical composition is a solid formulation, such as a freeze-dried or spray-dried composition that can be used as is, or a solid formulation that can be used when the physician or patient adds a solvent and / or diluent before use.Solid dosage forms can include tablets, such as compressed tablets, and / or coated tablets, and capsules (e.g., hard or soft gelatin capsules).The pharmaceutical composition can also be in the form of, for example, a sachet, dragee, powder, granule, lozenge, or powder for reconstitution.

[0152] The dosage forms may be immediate release, in which case they may include a water soluble or dispersible carrier, or they may be delayed, sustained, or modified release, in which case they may include a water insoluble polymer that modulates the dissolution rate of the dosage form in the digestive tract or under the skin.

[0153] In other embodiments, the pharmaceutical compositions may be delivered intranasally, bucally, or sublingually.

[0154] The pH of the aqueous formulation may be between pH 3 and pH 10. In one embodiment of the invention, the pH of the formulation is from about 7.0 to about 9.5. In another embodiment of the invention, the pH of the formulation is from about 3.0 to about 7.0.

[0155] In another embodiment of the invention, the pharmaceutical composition comprises a buffering agent. Non-limiting examples of buffering agents include arginine, aspartic acid, bicine, citrate, disodium hydrogen phosphate, fumaric acid, glycine, glycylglycine, histidine, lysine, maleic acid, malic acid, sodium acetate, sodium carbonate, sodium dihydrogen phosphate, sodium phosphate, succinic acid, tartaric acid, tricine, and tris(hydroxymethyl)aminomethane, and mixtures thereof. The buffering agents can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, for example, at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific buffering agents constitute alternative embodiments of the invention.

[0156] In another embodiment of the invention, the pharmaceutical composition comprises a preservative. Non-limiting examples of preservatives include benzethonium chloride, benzoic acid, benzyl alcohol, bronopol, butyl 4-hydroxybenzoate, chlorobutanol, chlorocresol, chlorhexidine, chlorphenesin, o-cresol, m-cresol, p-cresol, ethyl 4-hydroxybenzoate, imidurea, methyl 4-hydroxybenzoate, phenol, 2-phenoxyethanol, 2-phenylethanol, propyl 4-hydroxybenzoate, sodium dehydroacetate, thiomerosal, and mixtures thereof. The preservatives can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, for example, at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific preservatives constitute alternative embodiments of the invention.

[0157] In another embodiment of the present invention, the pharmaceutical composition comprises an isotonicity agent. Non-limiting examples of isotonicity agents include salts (e.g., sodium chloride), amino acids (e.g., glycine, histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, and threonine), alditols (e.g., glycerol, 1,2-propanediol propylene glycol), 1,3-propanediol, and 1,3-butanediol), polyethylene glycols (e.g., PEG400), and mixtures thereof. Another example of an isotonicity agent includes sugars. Non-limiting examples of sugars can be monosaccharides, disaccharides, or polysaccharides, or water-soluble glucans, such as fructose, glucose, mannose, sorbose, xylose, maltose, lactose, sucrose, trehalose, dextran, pullulan, dextrin, cyclodextrin, alpha and beta-HPCD, soluble starch, hydroxyethyl starch, and sodium carboxymethylcellulose. Another example of an isotonicity agent is a sugar alcohol, the term "sugar alcohol" being defined as a C(4-8) hydrocarbon having at least one -OH group. Non-limiting examples of sugar alcohols include mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, and arabitol. The isotonicity agent can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific isotonicity agents constitute alternative embodiments of the invention.

[0158] In another embodiment of the invention, the pharmaceutical composition comprises a chelating agent. Non-limiting examples of chelating agents include citric acid, aspartic acid, salts of ethylenediaminetetraacetic acid (EDTA), and mixtures thereof. The chelating agents can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, for example, about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific chelating agents constitute alternative embodiments of the invention.

[0159] In another embodiment of the invention, the pharmaceutical composition comprises a stabilizer, non-limiting examples of which include one or more aggregation inhibitors, one or more oxidation inhibitors, one or more surfactants, and / or one or more protease inhibitors.

[0160] In another embodiment of the invention, the pharmaceutical composition comprises a stabilizer, said stabilizer being carboxy- / hydroxycellulose and their derivatives (e.g., HPC, HPC-SL, HPC-L and HPMC), cyclodextrin, 2-methylthioethanol, polyethylene glycol (e.g., PEG3350), polyvinyl alcohol (PVA), polyvinylpyrrolidone, salts (e.g., sodium chloride), sulfur-containing substances, e.g., monothioglycerol), or thioglycolic acid. The stabilizers can be present individually or in aggregates at a concentration of about 0.01 mg / ml to about 50 mg / ml, e.g., about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these particular stabilizers constitute alternative embodiments of the invention.

[0161] In a further embodiment of the invention, the pharmaceutical composition comprises one or more surfactants, preferably one surfactant, at least one surfactant, or two different surfactants. The term "surfactant" refers to any molecule or ion composed of a water-soluble (hydrophilic) portion and a fat-soluble (lipophilic) portion. The surfactant may be selected, for example, from the group consisting of anionic surfactants, cationic surfactants, nonionic surfactants, and / or zwitterionic surfactants. The surfactants may be present individually or in aggregates at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific surfactants constitute alternative embodiments of the invention.

[0162] In further embodiments of the invention, the pharmaceutical composition comprises one or more protease inhibitors, such as EDTA, and / or benzamidine hydrochloride (HCl). The protease inhibitors can be present individually or in aggregates at a concentration of about 0.1 mg / ml to about 20 mg / ml. Pharmaceutical compositions comprising each one of these specific protease inhibitors constitute alternative embodiments of the invention.

[0163] In another general aspect, the invention relates to a method for producing a pharmaceutical composition comprising a monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof of the invention, comprising combining the monoclonal antibody or antigen-binding fragment thereof, and / or the bispecific antibody or antigen-binding fragment thereof with a pharma- ceutically acceptable carrier to obtain a pharmaceutical composition.

[0164] How to use In another general aspect, the present invention relates to a method of targeting IL13Ra2 on the surface of cancer cells in a subject to achieve cell killing, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof, that specifically binds to IL13Ra2, or a pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof of the present invention. Binding of an IL13Ra2 monoclonal or bispecific antibody or antigen-binding fragment to IL13Ra2 can mediate complement-dependent cytotoxicity (CDC), antibody-dependent cellular phagocytosis (ADCP), and / or antibody-dependent cellular cytotoxicity (ADCC) or other effects that result in the death of the targeted cancer cells. The monoclonal or bispecific antibody or antigen-binding fragment thereof can, for example, serve to recruit a conjugated drug and / or can form a bispecific antibody with another monoclonal antibody to mediate the death of the targeted cancer cells.

[0165] The functional activity of antibodies and antigen-binding fragments thereof that bind to IL13Ra2 can be characterized by methods known in the art and by methods described herein. Methods for characterizing antibodies and antigen-binding fragments thereof that bind to IL13Ra2 include, but are not limited to, affinity and specificity assays, including Biacore, ELISA, and OctetRed analysis, as well as detection of binding of antibodies and antigen-binding fragments to IL13Ra2 on cells (either cells transfected with IL13Ra2 or cells that naturally express IL13Ra2) by FACS. According to certain embodiments, methods for characterizing antibodies and antigen-binding fragments thereof that bind to IL13Ra2 include those described below.

[0166] In another general aspect, the present invention relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof that specifically binds IL13Ra2, or a pharmaceutical composition of the invention. The cancer can be selected from, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0167] In another general aspect, the present invention relates to a method of treating an inflammatory disease in a subject in need thereof comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof that specifically binds to IL13Ra2, or a pharmaceutical composition of the invention.

[0168] In another general aspect, the present invention relates to a method of treating an autoimmune disease in a subject in need thereof, comprising administering to the subject an isolated monoclonal antibody or antigen-binding fragment thereof, and / or a bispecific antibody or antigen-binding fragment thereof that specifically binds to IL13Ra2, or a pharmaceutical composition of the invention.

[0169] In another general aspect, the invention relates to a method of treating cancer in a subject in need thereof comprising administering to the subject a CAR-T cell and / or a CAR-NK cell of the invention. The cancer can be selected from, for example, but not limited to, lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0170] According to an embodiment of the invention, the CAR-T cells or CAR-NK cells comprise a therapeutically effective amount of the expressed CAR of the invention, and the pharmaceutical composition comprises a therapeutically effective amount of an anti-IL13Ra2 antibody or an antigen-binding fragment thereof. As used herein, the term "therapeutically effective amount" refers to an amount of an active ingredient or component that elicits a desired biological or pharmaceutical response in a subject. The therapeutically effective amount can be determined empirically and in a routine manner for the stated purpose.

[0171] As used herein with respect to a CAR, a therapeutically effective amount refers to an amount of a CAR molecule expressed in a transduced T cell or NK cell that modulates an immune response in a subject in need thereof. Additionally, as used herein with respect to a CAR, a therapeutically effective amount refers to an amount of a CAR molecule expressed in a transduced T cell or NK cell that provides treatment for a disease, disorder, or condition, prevents or slows the progression of a disease, disorder, or condition, or reduces or completely alleviates symptoms associated with a disease, disorder, or condition.

[0172] As used herein with respect to CAR-T cells or CAR-NK cells, a therapeutically effective amount refers to an amount of CAR-T cells or CAR-NK cells that modulates an immune response in a subject in need thereof. Additionally, as used herein with respect to CAR-T cells or CAR-NK cells, a therapeutically effective amount refers to an amount of CAR-T cells or CAR-NK cells that provides treatment for a disease, disorder, or condition, prevents or slows the progression of a disease, disorder, or condition, or reduces or completely alleviates symptoms associated with a disease, disorder, or condition.

[0173] As used herein with respect to an anti-IL13Ra2 antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to an amount of an anti-IL13Ra2 antibody or antigen-binding fragment thereof that modulates an immune response in a subject in need thereof, and as used herein with respect to an anti-IL13Ra2 antibody or antigen-binding fragment thereof, a therapeutically effective amount refers to an amount of an anti-IL13Ra2 antibody or antigen-binding fragment thereof that provides treatment of a disease, disorder or condition, prevents or slows the progression of a disease, disorder or condition, or reduces or completely alleviates symptoms associated with a disease, disorder or condition.

[0174] According to certain embodiments, the disease, disorder or condition to be treated is cancer, preferably a cancer selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0175] According to other particular embodiments, the disease, disorder or condition being treated is an inflammatory disease. According to other particular embodiments, the disease, disorder or condition being treated is an autoimmune disease.

[0176] According to certain embodiments, a therapeutically effective amount refers to an amount of therapy that is sufficient to achieve one, two, three, four or more of the following effects: (i) reducing or ameliorating the severity of the disease, disorder or condition being treated or symptoms associated therewith; (ii) reducing the duration of the disease, disorder or condition being treated or symptoms associated therewith; (iii) preventing the progression of the disease, disorder or condition being treated or symptoms associated therewith; (iv) causing regression of the disease, disorder or condition being treated or symptoms associated therewith; (v) preventing the development or onset of the disease, disorder or condition being treated or symptoms associated therewith. (vi) preventing the recurrence of the disease, disorder or condition to be treated or symptoms associated therewith; (vii) reducing hospitalization of a subject having the disease, disorder or condition to be treated or symptoms associated therewith; (viii) reducing the length of hospitalization of a subject having the disease, disorder or condition to be treated or symptoms associated therewith; (ix) increasing the survival of a subject having the disease, disorder or condition to be treated or symptoms associated therewith; (xi) inhibiting or reducing the disease, disorder or condition to be treated or symptoms associated therewith in a subject; and / or (xii) enhancing or improving the prophylactic or therapeutic efficacy of another therapy.

[0177] The therapeutically effective amount or dosage can vary depending on a variety of factors, such as the disease, disorder or condition to be treated, the means of administration, the target site, the physiological state of the subject (including, for example, age, weight, health status), whether the subject is human or animal, other pharmaceutical agents administered, and whether the treatment is prophylactic or therapeutic. Treatment dosages are optimally titrated to optimize safety and efficacy.

[0178] According to certain embodiments, the compositions described herein are formulated to be suitable for the intended route of administration to a subject. For example, the compositions described herein can be formulated to be suitable for intravenous, subcutaneous, or intramuscular administration.

[0179] The cells of the present invention can be administered in any convenient manner known to those skilled in the art. For example, the cells of the present invention can be administered to a subject by aerosol inhalation, injection, ingestion, transfusion, implantation and / or transplantation. The composition comprising the cells of the present invention can be administered intraarterially, subcutaneously, intradermally, intratumorally, intranodal, intramedullary, intramuscularly, intrapleurally, by intravenous (iv) injection, or intraperitoneally. In certain embodiments, the cells of the present invention can be administered with or without lymphodepletion of the subject.

[0180] Pharmaceutical compositions comprising the cells of the invention expressing a CAR of the invention can be provided in a sterile liquid formulation, typically an isotonic aqueous solution with a cell suspension, or optionally as an emulsion, dispersion, etc., which are typically buffered to a selected pH. The composition can include a carrier suitable for cell integrity and viability, as well as for administration of the cell composition, e.g., water, saline, phosphate buffered saline, etc.

[0181] Sterile injectable solutions can be prepared by incorporating the cells of the present invention in a suitable amount of a suitable solvent, optionally with various other ingredients. Such compositions can include pharma- ceutically acceptable carriers, diluents or excipients, such as sterile water, saline, glucose, dextrose, etc., suitable for use in the cell compositions and for administration to subjects, such as humans. Suitable buffers for providing cell compositions are well known in the art. Any vehicle, diluent or additive used is compatible with preserving the integrity and viability of the cells of the present invention.

[0182] The cells of the present invention can be administered in any physiologically acceptable vehicle. The population of cells containing the cells of the present invention can include a population of purified cells. One of skill in the art can easily determine the cells in a population of cells using a variety of well-known methods. The range of purity in a population of cells containing the genetically modified cells of the present invention can be about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 100%. The dosage can be easily adjusted by one of skill in the art, for example, a decrease in purity may require an increase in dosage.

[0183] The cells of the invention are generally administered as a dose based on the number of cells per kilogram of body weight (cells / kg) of the subject to whom the cells are administered. Generally, cell doses are about 10 per kg of body weight depending on the mode and location of administration. 4 ~about 10 10 Cell number, e.g., about 10 5 ~about 10 9 , about 10 5 ~about 10 8 , about 10 5 ~about 10 7 Or about 10 5 ~about 10 6 Generally, for systemic administration, higher doses are used than for local administration, where the immune cells of the invention are administered within the region of the tumor and / or cancer. Exemplary dose ranges include, but are not limited to, 1×10 4~1×10 8 、2×10 4 ~1×10 8 、3×10 4 ~1×10 8 、4×10 4 ~1×10 8 、5×10 4 ~6×10 8 、7×10 4 ~1×10 8 、8×10 4 ~1×10 8 、9×10 4 ~1×10 8 、1×10 5 ~1×10 8 、1×10 5 ~9×10 7 、1×10 5 ~8×10 7 、1×10 5 ~7×10 7 、1×10 5 ~6×10 7 、1×10 5 ~5×10 7 、1×10 5 ~4×10 7 、1×10 5 ~4×10 7 、1×10 5 ~3×10 7 、1×10 5 ~2×10 7 、1×10 5 ~1×10 7 、1×10 5 ~9×10 6 、1×10 5 ~8×10 6 、1×10 5 ~7×10 6 、1×10 5 ~6×10 6 、1×10 5 ~5×10 6 、1×10 5 ~4×10 6 、1×10 5 ~4×10 6 、1×10 5 ~3×10 6 、1×10 5 ~2×10 6 、1×105 ~1×10 6 , 2×10 5 ~9×10 7 , 2×10 5 ~8×10 7 , 2×10 5 ~7×10 7 , 2×10 5 ~6×10 7 , 2×10 5 ~5×10 7 , 2×10 5 ~4×10 7 , 2×10 5 ~4×10 7 , 2×10 5 ~3×10 7 , 2×10 5 ~2×10 7 , 2×10 5 ~1×10 7 , 2×10 5 ~9×10 6 , 2×10 5 ~8×10 6 , 2×10 5 ~7×10 6 , 2×10 5 ~6×10 6 , 2×10 5 ~5×10 6 , 2×10 5 ~4×10 6 , 2×10 5 ~4×10 6 , 2×10 5 ~3×10 6 , 2×10 5 ~2×10 6 , 2×10 5 ~1×10 6 , 3×10 5 ~3×10 6 The dosage can include, for example, the number of cells / kg, etc. Additionally, the dosage can be adjusted to account for whether a single dose or multiple doses are administered. The precise determination of what is considered an effective dose can be based on individual factors for each subject.

[0184] As used herein, the terms "treat", "treating" and "treatment" all refer to an improvement or reversal of at least one measurable physical parameter associated with cancer and / or an inflammatory or autoimmune disease, disorder or condition, which may be identifiable in the subject, but not necessarily in the subject. The terms "treat", "treating" and "treatment" can also refer to causing regression, preventing progression, or at least slowing progression of a disease, disorder or condition. In one particular embodiment, "treat", "treating" and "treatment" refer to alleviating, preventing the development or onset, or reducing the duration of one or more symptoms associated with a disease, disorder or condition, such as a tumor, more preferably a cancer. In one particular embodiment, "treat", "treating" and "treatment" refer to preventing the recurrence of a disease, disorder or condition. In one particular embodiment, "treat", "treating" and "treatment" refer to increasing the survival of a subject having a disease, disorder or condition. In one particular embodiment, "treat", "treating" and "treatment" refer to eliminating a disease, disorder or condition in a subject.

[0185] According to certain embodiments, compositions are provided for use in the treatment of cancer and / or inflammatory diseases, disorders or conditions. For cancer therapy, the compositions provided can be used in combination with another treatment, including, but not limited to, chemotherapy, anti-CD20 mAb, anti-TIM-3 mAb, anti-LAG-3 mAb, anti-EGFR mAb, anti-HER-2 mAb, anti-CD19 mAb, anti-CD33 mAb, anti-CD47 mAb, anti-CD73 mAb, anti-DLL-3 mAb, anti-Apelin mAb, anti-FOLR1 mAb, anti-CTLA-4 mAb, anti-PD-L1 mAb, anti-PD-1 mAb, anti-Claudin18.2 mAb, other immuno-oncology drugs, anti-angiogenesis drugs, radiation therapy, antibody-drug conjugates (ADC), targeted therapy, or other anti-cancer drugs. Antibodies against IL13Ra2 can be used to construct bispecific antibodies with partner mAbs against PD-1, PD-L1, LAG3, TIM-3, CTLA-4, EGFR, HER-2, CD19, CD20, CD33, CD73, CD47, CD3, Apelin, DLL-3, TIP-1, GPC3, Claudin18.2, folate receptor alpha (FOLR1), and / or any other TAA to treat cancers / tumors expressing both antigens. Two antibodies recognizing two different epitopes on IL13Ra2 can also be used to construct bispecific antibodies to treat cancers / tumors expressing IL13Ra2.

[0186] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that increases the efficacy of cells expressing a CAR molecule, including but not limited to, an antibody fragment that binds to CD73, CD39, PD1, PD-L1, PD-L2, CTLA4, TIM3 or LAG3, or an adenosine A2a receptor antagonist.

[0187] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that ameliorates one or more side effects associated with the administration of cells expressing a CAR molecule, including, but not limited to, a steroid, an inhibitor of TNFα, or an inhibitor of IL-6.

[0188] According to certain embodiments, a method of treating cancer in a subject in need thereof comprises administering to the subject the CAR-T cells and / or CAR-NK cells of the invention in combination with an agent that treats a disease associated with IL13Ra2, including, but not limited to, an anti-IL13Ra2 monoclonal or bispecific antibody.

[0189] As used herein, in the context of administering two or more therapies to a subject, the term "in combination" refers to the use of more than one therapy.The use of the term "in combination" does not limit the order in which the therapies are administered to a subject.For example, a first therapy (e.g., a composition described herein) can be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks before), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks after) the administration of a second therapy to a subject.

[0190] In another general aspect, the invention relates to a method for determining the level of IL13Ra2 in a subject, the method comprising: (a) obtaining a sample from the subject, (b) contacting the sample with a monoclonal antibody or antigen-binding fragment thereof of the invention, and (c) determining the level of IL13Ra2 in the subject.

[0191] As used herein, "sample" refers to a biological sample isolated from a subject, and can include, but is not limited to, whole blood, serum, plasma, blood cells, endothelial cells, tissue biopsy (e.g., cancer tissue), lymphatic fluid, peritoneal fluid, interstitial fluid, bone marrow, cerebrospinal fluid, saliva, mucus, sputum, sweat, urine, or any other secretion, excretion, or other bodily fluid. A "blood sample" refers to whole blood or any fraction thereof, including blood cells, serum, and plasma.

[0192] In certain embodiments, the level of IL13Ra2 in a subject can be determined using an assay selected from, but not limited to, Western blot assay, immunohistochemistry (IHC) and ELISA assay. Relative protein levels can be determined by utilizing Western blot analysis and IHC, and absolute protein levels can be determined by utilizing ELISA assay. When determining the relative level of IL13Ra2, the level of IL13Ra2 can be determined between at least two samples, for example, between samples from the same subject at different time points, between samples from different tissues in the same subject, and / or between samples from different subjects. Alternatively, when determining the absolute level of IL13Ra2, for example, by ELISA assay, the absolute level of IL13Ra2 in a sample can be determined by producing a standard for the ELISA assay before testing the sample. Those skilled in the art will understand the analytical techniques to be used to determine the level of IL13Ra2 in a sample from a subject using the antibody or antigen-binding fragment thereof of the present invention.

[0193] Utilizing the method for determining the level of IL13Ra2 in a sample from a subject can result in the diagnosis of abnormal (elevated, reduced or insufficient) IL13Ra2 levels in diseases and allow appropriate treatment decisions to be made. Such diseases can be selected from, but are not limited to, cancer, inflammatory diseases and autoimmune diseases. Furthermore, by monitoring the level of IL13Ra2 in a subject, the risk of developing such diseases can be determined based on knowledge of the level of IL13Ra2 in a particular disease and / or during the progression of a particular disease.

[0194] Embodiment The present invention also provides the following non-limiting embodiments.

[0195] In embodiment 1, (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

[0196] Embodiment 2 is an isolated monoclonal antibody or antigen-binding fragment of embodiment 1, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, or 169, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170.

[0197] In embodiment 3, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; or (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170. The isolated monoclonal antibody or antigen-binding fragment of embodiment 1 or 2, comprising:

[0198] Embodiment 4 is an isolated monoclonal antibody or antigen-binding fragment of any one of embodiments 1 to 3, which is chimeric or human or humanized.

[0199] Embodiment 5 is an isolated monoclonal antibody or antigen-binding fragment thereof according to embodiment 4, wherein the humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

[0200] In embodiment 6, The humanized monoclonal antibody or antigen-binding fragment thereof is (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213. The isolated monoclonal antibody or antigen-binding fragment thereof of embodiment 5, comprising:

[0201] Embodiment 7 is an isolated monoclonal antibody or antigen-binding fragment of any one of embodiments 1 to 6, which is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or mediating the recruitment of a conjugated drug, and / or forming a bispecific antibody with another mAb or antigen-binding fragment thereof that has cancer-killing activity.

[0202] Embodiment 8 is an isolated bispecific antibody or antigen-binding fragment thereof comprising the monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1 to 7.

[0203] Embodiment 9 is an isolated nucleic acid encoding the monoclonal antibody or antigen-binding fragment of any one of embodiments 1 to 7, or the bispecific antibody or antigen-binding fragment of embodiment 8.

[0204] Embodiment 10 is a vector comprising the isolated nucleic acid of embodiment 9.

[0205] Embodiment 11 is a host cell comprising the vector of embodiment 10.

[0206] Embodiment 12 is a pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 7, or the bispecific antibody or antigen-binding fragment thereof of Embodiment 8, and a pharma- ceutically acceptable carrier.

[0207]

[0036] Embodiment 13 is a method of targeting IL13Ra2 on the surface of cancer cells and / or treating cancer, treating an inflammatory disease, and / or treating an autoimmune disease in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of embodiment 12, optionally wherein the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0208] Embodiment 14 is a method for producing the monoclonal antibody or antigen-binding fragment of any one of Embodiments 1 to 7, or the bispecific antibody or antigen-binding fragment thereof of Embodiment 8, comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, under conditions for production of the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof, and recovering the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof from the cell or culture.

[0209] Embodiment 15 is a method for producing a pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1 to 7, or the bispecific antibody or antigen-binding fragment thereof of embodiment 8, comprising combining the monoclonal antibody or antigen-binding fragment thereof, or the bispecific antibody or antigen-binding fragment thereof with a pharma- ceutically acceptable carrier to obtain the pharmaceutical composition.

[0210] Embodiment 16 is a method for determining a level of IL13Ra2 in a subject, comprising: (1) obtaining a sample from a subject; (2) contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of embodiments 1 to 7; and (3) determining the level of IL13Ra2 in the subject; The method includes:

[0211] Embodiment 17 is the method of embodiment 16, wherein the sample is a tissue sample or a blood sample, optionally wherein the tissue sample is a cancer tissue sample.

[0212] Embodiment 18 is an isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to IL13Ra2; (b) hinge region; (c) a transmembrane domain, and (d) Intracellular signaling domain It is an isolated polynucleotide comprising:

[0213] Embodiment 19 is a method for producing an antigen-binding domain comprising the steps of: (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. and a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having a polypeptide sequence of:

[0214] Embodiment 20 is a method for the preparation of a heavy chain variable region having an antigen-binding domain having a polypeptide sequence at least 95% identical to SEQ ID NO: 1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, 169, 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212, or SEQ ID NO: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, 170, 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

[0215] Embodiment 21 is a method for determining whether the antigen-binding domain comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213; or (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213. 21. The isolated polynucleotide of embodiment 20, comprising:

[0216] Embodiment 22 is the isolated polynucleotide of any one of embodiments 18 to 21, wherein the antigen-binding domain is a single-chain variable fragment (scFv).

[0217] Embodiment 23 is the isolated polynucleotide of embodiment 22, wherein the single chain variable fragment (scFv) is humanized.

[0218] Embodiment 24 is the isolated polynucleotide of any one of embodiments 18 to 23, wherein the chimeric antigen receptor (CAR) comprises one or more antigen-binding domains, and / or the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0219] Embodiment 25 is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of any one of embodiments 18 to 24.

[0220] Embodiment 26 is a vector comprising the isolated polynucleotide of any one of embodiments 18 to 24.

[0221] Embodiment 27 is a host cell comprising the vector of embodiment 26, which is optionally a T cell or a NK cell, preferably a human T cell or a human NK cell.

[0222] Embodiment 28 is a method of making a host cell expressing a chimeric antigen receptor (CAR), comprising transducing a T cell or an NK cell with the vector of embodiment 26.

[0223]

[0041] Embodiment 29 is a method of producing chimeric antigen receptor (CAR)-T cells or chimeric antigen receptor (CAR)-NK cells, comprising culturing T cells or NK cells comprising an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of embodiments 18 to 24 under CAR-T cell or CAR-NK cell producing conditions, and recovering the CAR-T cells or CAR-NK cells.

[0224]

[0031] Embodiment 30 is a method of generating a cell comprising a chimeric antigen receptor (CAR), comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of embodiments 18 to 24, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0225]

[0036] Embodiment 31 is a method of treating cancer in a subject in need thereof comprising administering to a subject in need thereof the host cell of embodiment 27, optionally wherein the cancer is selected from lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

[0226]

[0036] Embodiment 32 is the method of embodiment 31, further comprising administering to a subject in need thereof an agent that increases the efficacy of the cells expressing a CAR, or an agent that ameliorates one or more side effects associated with administration of the cells expressing a CAR, or an agent that treats a disease associated with IL13Ra2.

[0227] [Example]

[0228] [Example 1] Identification of anti-IL13Ra2 monoclonal antibodies

[0229] Mice were immunized with recombinant human IL13Ra2 (ACROBiosystems; Cat#: IL2-H52H5, Lot#: C333P1-97HF1-Q5), hybridomas were generated and screened by ELISA and / or FACS. Positive clones were isolated and sequenced.

[0230] The sequences of the heavy and light chain variable regions (VH and VL regions, respectively) of the anti-IL13Ra2 monoclonal antibody are provided in Tables 1 and 2, and the CDR regions of the anti-IL13Ra2 monoclonal antibody are provided in Tables 3-6.

[0231] [Table 1-2]

[0232] [Table 2]

[0233] [Table 3]

[0234] [Table 4]

[0235] [Table 5]

[0236] [Table 6-1] [Table 6-2]

[0237] [Example 2] Production and purification of mAbs from culture medium of transfected cells To obtain recombinant anti-IL13Ra2 chimeric mAb, an expression vector containing the mouse variable regions (VH and VL) fused to the constant regions of human IgG1 heavy and kappa light chains, respectively, was transiently transfected into ExpiCHO-S cells. The recombinant antibody produced in suspension in ExpiCHO-S cells was purified using Protein A affinity chromatography.

[0238] [Example 3] ELISA binding analysis of purified chimeric mAbs The purified chimeric mAbs were tested in an ELISA assay for their ability to bind to immobilized IL13Ra2. Recombinant human IL13Ra2 in bicarbonate buffer (Sigma; St. Louis, MO; Cat#: C3041-100CAP) was coated onto 96-well plates for 1 h at room temperature. The plates are blocked with 5% BSA in TBST for 1 h at room temperature. Various concentrations of mAb were incubated in each well of each plate for 1 h at room temperature. The plates were washed and binding to IL13Ra2 was detected by incubating with mouse anti-human IgG Fc antibody conjugated to horseradish peroxidase (Genescript; Piscataway, NJ; Cat#: A01854) for 1 h at room temperature. Then, after washing, the ELISA was developed using One-Step Detection Solution (ThermoFisher Scientific; Waltham, MA; Cat#: 34028) and measured as absorbance at 450 nm. The results are shown in Figures 1A-1E. L3A6-C is a chimeric version of L3A6, and other antibodies follow the same naming convention.

[0239] [Example 4] FACS binding analysis of purified chimeric mAbs A375 cells were transferred to a 96-well plate. Approximately 50,000 cells were incubated with purified chimeric anti-IL13Ra2 mAb (variable regions of mouse mAb fused to human IgG1 heavy and kappa light chain constant regions, respectively) for 15 min at 4 °C. oThe cells were incubated at various concentrations in HBSS supplemented with 5% BSA and 0.05% sodium azide. The cells were then centrifuged for 5 min and washed once with FACS buffer (HBSS supplemented with 5% BSA and 0.05% sodium azide). The cells were then incubated with PE / Cyanine 7-labeled anti-human IgG Fc recombinant antibody (Biolegend; San Diego, CA; Cat# 366907) or F(ab')2-goat anti-human IgG Fc secondary antibody, Alexa Fluor™ 488 (ThermoFisher; Cat# H10120) and incubated for another 15 min on ice. The cells were then washed once in FACS buffer and resuspended in FACS buffer. The cells were then run on Attune NxT and the data was analyzed by Attune NxT software. The results are shown in Figures 2A-2B. L3A6-C is a chimeric version of L3A6, and other antibodies follow the same naming convention.

[0240] [Example 5] Humanization of anti-IL13Ra2 mAb The murine anti-IL13Ra2 mAb was humanized to reduce the possibility of immunogenicity when used in human patients. The sequences of the variable regions of the heavy and light chains (VH and VL) were compared to human antibody sequences in the Protein Data Bank (PDB) database, and a homology model was constructed. The CDRs of both the heavy and light chains of the murine mAb were grafted onto a human framework that was most likely to maintain the proper structure required for antigen binding. Back mutations or other mutations from human residues to murine residues were designed as necessary. The sequences of the humanized VH and VL regions are shown in Table 7. The humanized VH and VL regions were fused to the constant regions of human IgG1 heavy and kappa light chains, respectively. Binding of the humanized mAb to IL13Ra2 was evaluated using ELISA assays (Figures 3A-3H) and FACS assays (Figures 4A-4H). L3A6-H1L1 refers to a humanized mAb constructed using the L3A6-H1 heavy chain and the L3A6-L1 light chain shown in Table 7, and other humanized clones follow the same naming convention.

[0241] [Table 7-1] [Table 7-2] [Table 7-3]

[0242] [Example 6] Construction of a chimeric antigen receptor construct containing an anti-IL13Ra2 antigen-binding domain To construct the CAR construct, the mAb is converted to an scFv with a VH, VL, and (G4S)n linker and the scFv is fused to the N-terminus of the hinge and transmembrane domains derived from human CD8a (aa 114-118, Boursier JP et al., J. Biol. Chem. 268(3):2013-20 (1993)). The C-terminal intracellular signaling domain of the CAR is constructed by fusing the intracellular co-stimulatory domain of CD28 (aa 162-202, Aruffo A and Seed B, Proc. Natl. Acad. Sci. USA 84(23):8573-7 (1987)) followed by the activation domain from the CD3 zeta chain (aa 52-162, Letourneur F and Klausner RD, Proc. Natl. Acad. Sci. USA 88(20):8905-9 (1991)). The DNA sequence encoding the CAR is assembled and cloned into an expression vector (either retroviral, lentiviral, extrachromosomal or integrative) to generate the CAR construct using standard molecular biology cloning techniques.

[0243] Those skilled in the art will appreciate that changes may be made to the above-described embodiments without departing from the broad inventive concept thereof, and it is therefore understood that the invention is not limited to the particular embodiments disclosed, but is intended to cover modifications within the scope of the invention as defined herein.

Claims

1. (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, respectively, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, respectively, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively. An isolated monoclonal antibody or antigen-binding fragment thereof, comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having the polypeptide sequence of:

2. 2. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, comprising a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, or 169, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 2, 16, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, or 170.

3. (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; or (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

170.

3. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1 or 2, comprising:

4. 4. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 1, which is a chimeric or human or humanized monoclonal antibody or antigen-binding fragment thereof.

5. 5. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 4, wherein the humanized monoclonal antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NOs: 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

6. The humanized monoclonal antibody or antigen-binding fragment thereof, (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (13) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213, or (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

213.

6. The isolated monoclonal antibody or antigen-binding fragment thereof of claim 5, comprising:

7. 7. The isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 6, which is capable of inducing effector-mediated tumor cell lysis via antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC), and / or mediating the recruitment of a conjugated drug, and / or forming a bispecific antibody with another mAb or antigen-binding fragment thereof that has cancer-killing activity.

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

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

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

11. A host cell comprising the vector of claim 10.

12. 9. A pharmaceutical composition comprising an isolated monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, or a bispecific antibody or antigen-binding fragment thereof according to claim 8, and a pharma- ceutically acceptable carrier.

13. 13. A method of targeting IL13Ra2 on the surface of cancer cells and / or treating cancer and / or treating an inflammatory disease and / or treating an autoimmune disease in a subject in need thereof comprising administering to the subject the pharmaceutical composition of claim 12, wherein optionally the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, cholangiocarcinoma, cholangiocarcinoma, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

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

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

16. 1. A method for determining the level of IL13Ra2 in a subject, comprising: (a) obtaining a sample from a subject; (b) contacting the sample with the isolated monoclonal antibody or antigen-binding fragment thereof of any one of claims 1 to 7; and (c) determining the level of IL13Ra2 in the subject A method comprising:

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

18. An isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), the CAR comprising: (a) an extracellular domain comprising at least one antigen-binding domain that specifically binds to IL13Ra2; (b) hinge region; (c) a transmembrane domain, and (d) Intracellular signaling domain 2. An isolated polynucleotide comprising:

19. The antigen-binding domain comprises: (1) SEQ ID NOs: 3, 4, 5, 6, 7, and 8, or SEQ ID NOs: 9, 10, 11, 12, 13, and 14, respectively; (2) SEQ ID NOs: 17, 18, 19, 20, 21, and 22, respectively, or SEQ ID NOs: 23, 24, 25, 26, 27, and 28, respectively; (3) SEQ ID NOs: 31, 32, 33, 34, 35, and 36, or SEQ ID NOs: 37, 38, 39, 40, 41, and 42, respectively; (4) SEQ ID NOs: 45, 46, 47, 48, 49, and 50, or SEQ ID NOs: 51, 52, 53, 54, 55, and 56, respectively; (5) SEQ ID NOs: 59, 60, 61, 62, 63, and 64, or SEQ ID NOs: 65, 66, 67, 68, 69, and 70, respectively; (6) SEQ ID NOs: 73, 74, 75, 76, 77, and 78, or SEQ ID NOs: 79, 80, 81, 82, 83, and 84, respectively; (7) SEQ ID NOs: 87, 88, 89, 90, 91 and 92, or SEQ ID NOs: 93, 94, 95, 96, 97 and 98, respectively; (8) SEQ ID NOs: 101, 102, 103, 104, 105 and 106, or SEQ ID NOs: 107, 108, 109, 110, 111 and 112, respectively; (9) SEQ ID NOs: 115, 116, 117, 118, 119, and 120, respectively, or SEQ ID NOs: 121, 122, 123, 124, 125, and 126, respectively; (10) SEQ ID NOs: 129, 130, 131, 132, 133, and 134, or SEQ ID NOs: 135, 136, 137, 138, 139, and 140, respectively; (11) SEQ ID NOs: 143, 144, 145, 146, 147, and 148, or SEQ ID NOs: 149, 150, 151, 152, 153, and 154, respectively; (12) SEQ ID NOs: 157, 158, 159, 160, 161, and 162, respectively, or SEQ ID NOs: 163, 164, 165, 166, 167, and 168, respectively; or (13) SEQ ID NOs: 171, 172, 173, 174, 175, and 176, respectively, or SEQ ID NOs: 177, 178, 179, 180, 181, and 182, respectively.

19. The isolated polynucleotide of claim 18, comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 having a polypeptide sequence of:

20. a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 1, 15, 29, 43, 57, 71, 85, 99, 113, 127, 141, 155, 169, 183, 184, 185, 186, 191, 192, 193, 197, 198, 199, 203, 204, 207, 208, 211, or 212, or a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 2, 20. The isolated polynucleotide of claim 18 or 19, comprising a light chain variable region having a polypeptide sequence that is at least 95% identical to 6, 30, 44, 58, 72, 86, 100, 114, 128, 142, 156, 170, 187, 188, 189, 190, 194, 195, 196, 200, 201, 202, 205, 206, 209, 210, or 213.

21. The antigen-binding domain comprises: (1) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 2; (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16; (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 29 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 30; (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 43 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 44; (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 57 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 58; (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 71 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 72; (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 85 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 86; (8) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 99 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 100; (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 114; (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 127 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 128; (11) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 141 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 142; (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 155 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 156; (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 169 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 170; (14) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (15) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (16) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (17) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (18) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (19) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (21) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (22) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (23) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (24) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (25) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 185 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (26) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187; (27) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 188; (28) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 189; (29) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 186 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190; (30) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (31) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (32) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 191 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (33) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (34) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (35) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 192 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (36) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 194; (37) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195; (38) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 193 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196; (39) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (40) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (41) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 197 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (42) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (43) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (44) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 198 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (45) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 200; (46) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 201; (47) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 199 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 202; (48) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (49) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 203 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (50) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 205; (51) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 204 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 206; (52) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (53) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 207 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (54) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 209; (55) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 208 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 210; (56) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 211 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 213; or (57) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 212 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

213.

21. The isolated polynucleotide of claim 20, comprising:

22. 22. The isolated polynucleotide of any one of claims 18 to 21, wherein the antigen-binding domain is a single-chain variable fragment (scFv).

23. 23. The isolated polynucleotide of claim 22, wherein the single chain variable fragment (scFv) is humanized.

24. 24. The isolated polynucleotide of any one of claims 18 to 23, wherein the chimeric antigen receptor (CAR) comprises one or more antigen binding domains and / or the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

25. 25. A chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of any one of claims 18 to 24.

26. 25. A vector comprising the isolated polynucleotide of any one of claims 18 to 24.

27. 27. A host cell comprising the vector of claim 26, optionally a T cell or a NK cell, preferably a human T cell or a human NK cell.

28. 27. A method of making a host cell expressing a chimeric antigen receptor (CAR), comprising transducing a T cell or a NK cell with the vector of claim 26.

29. 25. A method for producing chimeric antigen receptor (CAR)-T cells or chimeric antigen receptor (CAR)-NK cells, comprising culturing T cells or NK cells comprising an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) of any one of claims 18 to 24 under CAR-T cell or CAR-NK cell producing conditions, and recovering the CAR-T cells or CAR-NK cells.

30. 25. A method of generating a cell comprising a chimeric antigen receptor (CAR), comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid encoding the chimeric antigen receptor (CAR) of any one of claims 18 to 24, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

31. 28. A method of treating cancer in a subject in need thereof comprising administering to a subject in need thereof the host cell of claim 27, optionally wherein the cancer is selected from lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma, mesothelioma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), multiple myeloma (MM), acute myelogenous leukemia (AML) and other liquid tumors.

32. The method of claim 31, further comprising administering to a subject in need thereof an agent that increases the efficacy of the cell expressing a CAR, or an agent that ameliorates one or more side effects associated with administration of the cell expressing a CAR, or an agent that treats a disease associated with IL13Ra2.

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