Antibodies against VEGF and methods of use thereof

Abstract

The present invention is directed to human monoclonal antibodies that bind to Vascular Endothelial Growth Factor (VEGF) protein and methods of use thereof.

Description

Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 ANTIBODIES AGAINST VEGF AND METHODS OF USE THEREOF

[0001] This application claims priority to U.S. Provisional Application No.63 / 563,120 filed on March 08, 2024, the entire contents of each of which are incorporated herein by reference. Sequence Listing

[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on [ ], is named [ ] and is [ ] bytes in size. Background

[0003] The TME consists of pro-angiogenic growth factors, such as vascular endothelial growth factor (VEGF), that contribute to the abnormal vasculature of tumor cells. The VEGF family is led by VEGF-A, a signaling protein responsible for blood vessel formation, and its receptor VEGFR-2. The VEGF-A / VEGFR-2 axis activates angiogenesis and promotes tumor development. The up-regulation of VEGF-A produces a structurally and functionally abnormal tumor vasculature with features such as irregular branching patterns. The abnormal vasculature limits the egress of therapeutic cells out of the tumor vasculature and into the tumor tissue which reduces the efficiency of drug delivery. Summary of the invention

[0004] Aspects are directed to an antibody or antigen-binding fragment or variant thereof that specifically binds Vascular Endothelial Growth Factor (VEGF) protein.

[0005] In embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the HCVR comprises the complementarity determining regions (CDRs) HCDR1, HCDR2 and HCDR3 and the LCVR comprises CDRs LCDR1, LCDR2 and LCDR3, wherein: the amino acid sequence of HCDR1 is GFTFNDYE, the amino acid sequence of HCDR2 is VTARGGTE, the amino acid sequence of HCDR3 is ARSARDSYGGDYFDY, the amino acid sequence of LCDR1 is NIGSKS, the amino acid sequence of LCDR2 is YDS, and the amino acid sequence of LCDR3 isDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 QVWDSSSDHPV; or the amino acid sequence of HCDR1 is GFAFRTSA, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ASSSGYFDDAFDI, the amino acid sequence of LCDR1 is NIGSKS, the amino acid sequence of LCDR2 is DDS, and the amino acid sequence of LCDR3 is QVWDGGNDDHVV; or the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVSPY, the amino acid sequence of LCDR2 is GAS, and the amino acid sequence of LCDR3 is QQYGSSPLT; or the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVGEN, the amino acid sequence of LCDR2 is AAS, and the amino acid sequence of LCDR3 is QQFGRSPLT; or the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVSRS, the amino acid sequence of LCDR2 is DAS, and the amino acid sequence of LCDR3 is QQYGSSPLT; or the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVRNN, the amino acid sequence of LCDR2 is SAS, and the amino acid sequence of LCDR3 is QQYGSSPIT; or the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is SSYTITQHRG; or the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is ISYTITSIVV; or the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is SSYTITSIVV; or the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is IYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDN, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acidDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 sequence of LCDR3 is SSYTITSIVV; or the amino acid sequence of HCDR1 is GGTFKDYS, the amino acid sequence of HCDR2 is IIPMYGST, the amino acid sequence of HCDR3 is AREEEGLYRAFDL, the amino acid sequence of LCDR1 is SIGSKS, the amino acid sequence of LCDR2 is DDT, and the amino acid sequence of LCDR3 is QVWDSSSDHPV; or the amino acid sequence of HCDR1 is GFTFSSYS, the amino acid sequence of HCDR2 is ISSSGTYI, the amino acid sequence of HCDR3 is ARDSSSWSRTGGMDV, the amino acid sequence of LCDR1 is SLRRYY, the amino acid sequence of LCDR2 is GKN, and the amino acid sequence of LCDR3 is NSRDSSGNPVV; or the amino acid sequence of HCDR1 is GYVFTNYF, the amino acid sequence of HCDR2 is INPNSGGT, the amino acid sequence of HCDR3 is ARERKRGYSYGPLDY, the amino acid sequence of LCDR1 is SLSNFF, the amino acid sequence of LCDR2 is GKN, and the amino acid sequence of LCDR3 is NSRDSNGDLVV; or the amino acid sequence of HCDR1 is GFTFDDYA, the amino acid sequence of HCDR2 is LSWNSGSV, the amino acid sequence of HCDR3 is AKGTKDSGASGPYFFDY, the amino acid sequence of LCDR1 is TGAVTSDNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGARV; or the amino acid sequence of HCDR1 is GFSFNNYA, the amino acid sequence of HCDR2 is ISWNSGSI, the amino acid sequence of HCDR3 is AKDPAKFRDGYNSDGFDI, the amino acid sequence of LCDR1 is TGAVTSAFY, the amino acid sequence of LCDR2 is NTD, and the amino acid sequence of LCDR3 is LLSYSGARPV; or the amino acid sequence of HCDR1 is GFIFDDYA, the amino acid sequence of HCDR2 is INWISGSI, the amino acid sequence of HCDR3 is ARDNEGLYRGFDY, the amino acid sequence of LCDR1 is GSNIGAPYD, the amino acid sequence of LCDR2 is GDN, and the amino acid sequence of LCDR3 is QSYDSSLSGHVV; or the amino acid sequence of HCDR1 is ISGDSVSSGA, the amino acid sequence of HCDR2 is TYYSSKWYN, the amino acid sequence of HCDR3 is GRTLSALGNNWFDP, the amino acid sequence of LCDR1 is TGAVTSDNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGARV; or the amino acid sequence of HCDR1 is GFDFSNWG, the amino acid sequence of HCDR2 is IGHDGMSQ, the amino acid sequence of HCDR3 is ARDLNSGYTDR, the amino acid sequence of LCDR1 is TGTVTSTNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGSGV; or the amino acid sequence of HCDR1 is GYTFSNYG, the amino acid sequence of HCDR2 is ISGHNGKT, the amino acid sequence of HCDR3 is ARDRMRGLYGMDV, the amino acid sequence of LCDR1 is TGAVTSGNY, the amino acid sequence of LCDR2 is IQP, and theDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 amino acid sequence of LCDR3 is LSYPGARG; or the amino acid sequence of HCDR1 is GGTFSSYA, the amino acid sequence of HCDR2 is IIPIFGTA, the amino acid sequence of HCDR3 is ARILGGGMDV, the amino acid sequence of LCDR1 is QSVSSSY, the amino acid sequence of LCDR2 is GAS, and the amino acid sequence of LCDR3 is QQYGSSPWT; or the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; or the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; or the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; or the amino acid sequence of HCDR1 is GYRFITYW, the amino acid sequence of HCDR2 is IYPGDSES, the amino acid sequence of HCDR3 is ARRMGGNDAFDV, the amino acid sequence of LCDR1 is QSISSY, the amino acid sequence of LCDR2 is AAS, and the amino acid sequence of LCDR3 is QQANSLRWT; or the amino acid sequence of HCDR1 is GFSFSSYP, the amino acid sequence of HCDR2 is IRSGGSPI, the amino acid sequence of HCDR3 is ARDDLYAFDI, the amino acid sequence of LCDR1 is QDISNY, the amino acid sequence of LCDR2 is DAS, and the amino acid sequence of LCDR3 is QQSYSTPIT; or the amino acid sequence of HCDR1 is GFIFNNYW, the amino acid sequence of HCDR2 is IERYGGEI, the amino acid sequence of HCDR3 is ARPSWNSGSYFDY, the amino acid sequence of LCDR1 is GSNIGTYT, the amino acid sequence of LCDR2 is RNS, and the amino acid sequence of LCDR3 is SAWDDSLGGEV; or the amino acid sequence of HCDR1 is GFIFNNYW, the amino acid sequence of HCDR2 is IERYGGEI, the amino acid sequence of HCDR3 is ARPSWNSGSYFDY, the amino acid sequence of LCDR1 is NSNIGSYT, the amino acid sequence of LCDR2 is SNN, and the amino acid sequence of LCDR3 is SAWDDSLGGEV; or the amino acid sequence of HCDR1 is GFTFSDYW, the amino acid sequence of HCDR2 is IERYGAEE, the amino acid sequence of HCDR3 is ARPTFNSGSYFDY, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and theDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 amino acid sequence of LCDR3 is SAWDDSLGGEV; or the amino acid sequence of HCDR1 is GFSLTTRGLA, the amino acid sequence of HCDR2 is VYWDDDK, the amino acid sequence of HCDR3 is AHFGYYYFDS, the amino acid sequence of LCDR1 is SGSIASSY, the amino acid sequence of LCDR2 is EDN, and the amino acid sequence of LCDR3 is QSYGSGNWV; or the amino acid sequence of HCDR1 is GFSLTTRGLA, the amino acid sequence of HCDR2 is VYWDDDK, the amino acid sequence of HCDR3 is AHFGYYYFDS, the amino acid sequence of LCDR1 is SGSIASNF, the amino acid sequence of LCDR2 is EDK, and the amino acid sequence of LCDR3 is QSYDTSTHWV; or the amino acid sequence of HCDR1 is GFSFRMFG, the amino acid sequence of HCDR2 is ISYDGSND, the amino acid sequence of HCDR3 is ARDLGLLQYYYYGMDV, the amino acid sequence of LCDR1 is SSDVGGYNY, the amino acid sequence of LCDR2 is DVS, and the amino acid sequence of LCDR3 is SSYTSSSTLNWV; or the amino acid sequence of HCDR1 is GFSFTMFG, the amino acid sequence of HCDR2 is ISYDGSND, the amino acid sequence of HCDR3 is ARDLGLLQYYYYGMDV, the amino acid sequence of LCDR1 is SSDVGGYNY, the amino acid sequence of LCDR2 is DVS, and the amino acid sequence of LCDR3 is SSYTSSSTLNWG.

[0006] In embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the amino acid sequence of the LCVR is SYELTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTIS RVEAGDEADYYCQVWDSSSDHPVFGGGTKLAVL and the amino acid sequence of the HCVR is QVQLVQSGGALVRPGGSLRLSCVASGFTFNDYEMNWVRQAPGTGLEWVSSVTARGGTEYYADSVKGRFTISRDNS KNTVYLQMNSLRGEDTAVYYCARSARDSYGGDYFDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPAQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTIS RVEAGDEADYYCQVWDGGNDDHVVLGGETKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGSVQPGGSLTLSCAASGFAFRTSAMTWVRQAPGRGLEWMGIVYPGDSDTIYSPSFRGQVTISADKS ISTAYLQWSSLKASDTAMYYCASSSGYFDDAFDIWGQGTMVTVSS; or the amino acid sequence of the LCVR is EIVLTQSPATLSLSPGERATLSCRAGQSVSPYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKSDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the amino acid sequence of the LCVR is ETTLTQSPVTLSVSPGERVTLSCRASQSVGENLAWYQQKPGQAPRLLMYAASTRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQFGRSPLTFGGGTKVEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISVDKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the amino acid sequence of the LCVR is EIVLTQSPGTLSLSPGERATLSCRASQSVSRSLAWYQQKPGQAPRLLIYDASTRATGIPARFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the amino acid sequence of the LCVR is EIVLTQSPATLSLSPGERATLSCRASQSVRNNLAWYQQKPGQAPRLLIYSASSRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPITFGQGTRLEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCSSYTITQHRGFGGGTKLTVP and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCISYTITSIVVFGGETKLTVL and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASLDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 TISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISVDKS INTAYLQWNSLKASDTALYFCARQKWETMTASAFDNWGQGTTVTVSS; or the amino acid sequence of the LCVR is SYELTHPPSVSVAPGKTATMTCGGDSIGSKSLNWYQQKPGQAPVLVVYDDTDRPSGIPERFSGSNSGDTATLTLS RVEAGDEADYYCQVWDSSSDHPVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVTKPGSSVKVSCRTSGGTFKDYSFSWVRQAPGRGLEWMGGIIPMYGSTDYTKKFQGRVTLTADTS TTTLYMELSSLRSEDTAVYYCAREEEGLYRAFDLWGQGTMVTVSS; or the amino acid sequence of the LCVR is SSELTQDPAVSVALGQTVRITCQGDSLRRYYASWYQQKPGQAPVLVFYGKNTRPSGIPDRISGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNPVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSGTYIYYADSVKGRFTISRDNA KNSLYLQMNSLGAEDTAVYYCARDSSSWSRTGGMDVWGQGTTVTVSS; or the amino acid sequence of the LCVR is SSELTQDPAVSVALGQTVRITCQGDSLSNFFAGWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSNGDLVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGSELKKPGAPVKVSCKTSGYVFTNYFIHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVTMTRDTS ISTAYMELSRLRSDDTAVYYCARERKRGYSYGPLDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is QTVVTQDPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVLGGKAAL TLSGAQPEDEAEYYCLLSYSGARVFGGETKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVPGRSLRLSCAASGFTFDDYAVHWVRQAPGKGLEWVSGLSWNSGSVGYADSVRGRFTISRDNAG NSLYLQMNGLRAEDTAFYYCAKGTKDSGASGPYFFDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGAVTLTCASSTGAVTSAFYPNWFQQKPGQAPKALIYNTDNKHSWTPARFSGSLLGGKAAL TLSGAQPEDEAEYYCLLSYSGARPVFGGGTKLTVL and the amino acid sequence of the HCVR is EVQLVESGGGLVQAGGSLTLSCAASGFSFNNYAMSWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRDNA KNSLYLQMNTLRAEDTALYYCAKDPAKFRDGYNSDGFDIWGQGTTVTVSS; or the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGTGSNIGAPYDVHWYQHLPGTAPKLLIYGDNNRPSGVPDRFSGSKSGTSPSL AISGLRSEDEADYYCQSYDSSLSGHVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVQPGRSLRLSCAASGFIFDDYAMHWVRQAPGKGLEWVSGINWISGSIGYADSVKGRFTVSRDNA KNSLYLQMNSLRSEDTAVYYCARDNEGLYRGFDYWGQGTLVTVSS; or the amino acid sequence of theDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 LCVR is QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVLGGKAAL TLSGAQPEDEAEYYCLLSYSGARVFGGGTKLTVL and the amino acid sequence of the HCVR is MAQVQLQQSGPGLVKPSQTLSLTCGISGDSVSSGAWNWIRSPSRGLQWLGRTYYSSKWYNDYAESVKSRISINAD TSKNQFSLHLNSVTPEDTAVYYCGRTLSALGNNWFDPWGQGTLVTVSS; or the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGTVTLTCTSSTGTVTSTNYPNWFQQKPGQAPRPLIYSTTKRHSWTPARFSGSLLGGKAAL TLSGAQPEDEAEYFCLLSYSGSGVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFDFSNWGMHWIRQAPGKGLEGVAVIGHDGMSQRYADSVKGRFTVSRDNS KNTQYLEMNSLRVEDTALYYCARDLNSGYTDRWGQGTLVTVSS; or the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSGNYPNWFQQKPGQAPRPLIYIQPKDTPGPLPGSQAPPWGQSCPD TVRCAPEDEADYYCLLSYPGARGLRTGTNLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVNKPGASVKVSCKASGYTFSNYGLTWMRQAPGQGLEWMGWISGHNGKTLSAQKFQDRLLMTTDTS TTTAWLELRSLRSDDTAVYYCARDRMRGLYGMDVWGQGTMVTVSS; or the amino acid sequence of the LCVR is EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLT ISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK and the amino acid sequence of the HCVR is QVQLQQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADES TSTAYMELSSLRSEDTAVYYCARILGGGMDVWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSVLTHPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the amino acid sequence of the HCVR is VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the amino acid sequence of the HCVR is VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGETKLTVL and the amino acid sequence of the HCVR isDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the amino acid sequence of the LCVR is DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQQANSLRWTFGQGTKVEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLRISCQGSGYRFITYWIGWVRQTPGKGLEWMGAIYPGDSESTYSPPFQGQVTMSVDKS INTAYLQWSSLKASDTATYYCARRMGGNDAFDVWGQGTLVTVSS; or the amino acid sequence of the LCVR is DIVMTQTPPSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFTGSGSGTDFTLTI SSLQPEDFATYYCQQSYSTPITFGQGTRLEIK and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCVASGFSFSSYPMNWVRQAPGKGLEWISHIRSGGSPISYADSVKGRFTISRDNA KNSLYLQMNSLRAEDTGVYYCARDDLYAFDIWGQGTMVTVSS; or the amino acid sequence of the LCVR is LPVLTQPPSASGTPGQRVTISCSGSGSNIGTYTVNWYQQLPGTAPKLLIYRNSQRPSGVPARFSASKSGTSASLA ISGLRSEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFTISRDNA KNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is LPVLTQPPSASGTPGQRVTISCSGSNSNIGSYTVNWYQQFPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLA ISGLRFEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFTISRDNA KNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYFSAWDDSLGGEVFGTGTKVNVL and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCEASGFTFSDYWMGWVRQAPGKGLEWVANIERYGAEEKYVDSVRGRFTISRDNA KNLLYLHMDSLRAEDTAMYYCARPTFNSGSYFDYWGQGTLVTVSS; or the amino acid sequence of the LCVR is NFMLTQPHSVSESPGETVTISCTRSSGSIASSYVQWYQQRPGSSPTTVVYEDNQRPSGVPDRFSGSIDSSSNSAS LTISGLKTEDEADYYCQSYGSGNWVFGGGTKLTVL and the amino acid sequence of the HCVR is QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRLSISKDS SKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or the amino acid sequence of the LCVR isDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 NFMLTHPHSVSGSPGETVTISCTRNSGSIASNFVQWYQQRPGGSPKNVIHEDKRRPSGVPDRFSGSIDTSSNSAF LTISGLKTDDEADYYCQSYDTSTHWVFGGGTRLTVL and the amino acid sequence of the HCVR is QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRLSISKDS SKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNRFSGSKSGNTASL TISGLQAEDEADYYCSSYTSSSTLNWVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFSFRMFGLHWVRQAPGKGLEWVAFISYDGSNDYYADSVKGRFTISRDNS KNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS; or the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNLFSGSKSGNTASL TISGLQAEDEADYYCSSYTSSSTLNWGCGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFSFTMFGLHWVRQAPGKGLEWVAFISYDGSNDYYAHSVKGRFTISRDNS KNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS.

[0007] In embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SYELTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTIS RVEAGDEADYYCQVWDSSSDHPVFGGGTKLAVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGALVRPGGSLRLSCVASGFTFNDYEMNWVRQAPGTGLEWVSSVTARGGTEYYADSVKGRFTISRDNS KNTVYLQMNSLRGEDTAVYYCARSARDSYGGDYFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPAQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTIS RVEAGDEADYYCQVWDGGNDDHVVLGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGSVQPGGSLTLSCAASGFAFRTSAMTWVRQAPGRGLEWMGIVYPGDSDTIYSPSFRGQVTISADKS ISTAYLQWSSLKASDTAMYYCASSSGYFDDAFDIWGQGTMVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPATLSLSPGERATLSCRAGQSVSPYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequenceDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence ETTLTQSPVTLSVSPGERVTLSCRASQSVGENLAWYQQKPGQAPRLLMYAASTRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQFGRSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISVDKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPGTLSLSPGERATLSCRASQSVSRSLAWYQQKPGQAPRLLIYDASTRATGIPARFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPATLSLSPGERATLSCRASQSVRNNLAWYQQKPGQAPRLLIYSASSRATGIPDRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPITFGQGTRLEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVTISADKS ISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCSSYTITQHRGFGGGTKLTVP and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCISYTITSIVVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the VL region comprises anDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVTISADRS TNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASL TISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISVDKS INTAYLQWNSLKASDTALYFCARQKWETMTASAFDNWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SYELTHPPSVSVAPGKTATMTCGGDSIGSKSLNWYQQKPGQAPVLVVYDDTDRPSGIPERFSGSNSGDTATLTLS RVEAGDEADYYCQVWDSSSDHPVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVTKPGSSVKVSCRTSGGTFKDYSFSWVRQAPGRGLEWMGGIIPMYGSTDYTKKFQGRVTLTADTS TTTLYMELSSLRSEDTAVYYCAREEEGLYRAFDLWGQGTMVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SSELTQDPAVSVALGQTVRITCQGDSLRRYYASWYQQKPGQAPVLVFYGKNTRPSGIPDRISGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNPVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSGTYIYYADSVKGRFTISRDNA KNSLYLQMNSLGAEDTAVYYCARDSSSWSRTGGMDVWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SSELTQDPAVSVALGQTVRITCQGDSLSNFFAGWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSNGDLVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGSELKKPGAPVKVSCKTSGYVFTNYFIHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVTMTRDTS ISTAYMELSRLRSDDTAVYYCARERKRGYSYGPLDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQDPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVLGGKAALDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 TLSGAQPEDEAEYYCLLSYSGARVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVPGRSLRLSCAASGFTFDDYAVHWVRQAPGKGLEWVSGLSWNSGSVGYADSVRGRFTISRDNAG NSLYLQMNGLRAEDTAFYYCAKGTKDSGASGPYFFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGAVTLTCASSTGAVTSAFYPNWFQQKPGQAPKALIYNTDNKHSWTPARFSGSLLGGKAAL TLSGAQPEDEAEYYCLLSYSGARPVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVESGGGLVQAGGSLTLSCAASGFSFNNYAMSWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRDNA KNSLYLQMNTLRAEDTALYYCAKDPAKFRDGYNSDGFDIWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGTGSNIGAPYDVHWYQHLPGTAPKLLIYGDNNRPSGVPDRFSGSKSGTSPSL AISGLRSEDEADYYCQSYDSSLSGHVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVQPGRSLRLSCAASGFIFDDYAMHWVRQAPGKGLEWVSGINWISGSIGYADSVKGRFTVSRDNA KNSLYLQMNSLRSEDTAVYYCARDNEGLYRGFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVLGGKAAL TLSGAQPEDEAEYYCLLSYSGARVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence MAQVQLQQSGPGLVKPSQTLSLTCGISGDSVSSGAWNWIRSPSRGLQWLGRTYYSSKWYNDYAESVKSRISINAD TSKNQFSLHLNSVTPEDTAVYYCGRTLSALGNNWFDPWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCTSSTGTVTSTNYPNWFQQKPGQAPRPLIYSTTKRHSWTPARFSGSLLGGKAAL TLSGAQPEDEAEYFCLLSYSGSGVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFDFSNWGMHWIRQAPGKGLEGVAVIGHDGMSQRYADSVKGRFTVSRDNS KNTQYLEMNSLRVEDTALYYCARDLNSGYTDRWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSGNYPNWFQQKPGQAPRPLIYIQPKDTPGPLPGSQAPPWGQSCPD TVRCAPEDEADYYCLLSYPGARGLRTGTNLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequenceDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 QVQLVQSGAEVNKPGASVKVSCKASGYTFSNYGLTWMRQAPGQGLEWMGWISGHNGKTLSAQKFQDRLLMTTDTS TTTAWLELRSLRSDDTAVYYCARDRMRGLYGMDVWGQGTMVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLT ISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLQQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADES TSTAYMELSSLRSEDTAVYYCARILGGGMDVWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTHPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYYCQSYDSSLSGWVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTLIADEST RTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQQANSLRWTFGQGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLRISCQGSGYRFITYWIGWVRQTPGKGLEWMGAIYPGDSESTYSPPFQGQVTMSVDKS INTAYLQWSSLKASDTATYYCARRMGGNDAFDVWGQGTLVTVSS; or the VL region comprises an aminoDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 acid sequence that has at least 95% sequence identity to the amino acid sequence DIVMTQTPPSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFTGSGSGTDFTLTI SSLQPEDFATYYCQQSYSTPITFGQGTRLEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCVASGFSFSSYPMNWVRQAPGKGLEWISHIRSGGSPISYADSVKGRFTISRDNA KNSLYLQMNSLRAEDTGVYYCARDDLYAFDIWGQGTMVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence LPVLTQPPSASGTPGQRVTISCSGSGSNIGTYTVNWYQQLPGTAPKLLIYRNSQRPSGVPARFSASKSGTSASLA ISGLRSEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFTISRDNA KNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence LPVLTQPPSASGTPGQRVTISCSGSNSNIGSYTVNWYQQFPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLA ISGLRFEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFTISRDNA KNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL AITGLQAEDEADYFSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCEASGFTFSDYWMGWVRQAPGKGLEWVANIERYGAEEKYVDSVRGRFTISRDNA KNLLYLHMDSLRAEDTAMYYCARPTFNSGSYFDYWGQGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence NFMLTQPHSVSESPGETVTISCTRSSGSIASSYVQWYQQRPGSSPTTVVYEDNQRPSGVPDRFSGSIDSSSNSAS LTISGLKTEDEADYYCQSYGSGNWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRLSISKDS SKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence NFMLTHPHSVSGSPGETVTISCTRNSGSIASNFVQWYQQRPGGSPKNVIHEDKRRPSGVPDRFSGSIDTSSNSAFDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 LTISGLKTDDEADYYCQSYDTSTHWVFGGGTRLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRLSISKDS SKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNRFSGSKSGNTASL TISGLQAEDEADYYCSSYTSSSTLNWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFSFRMFGLHWVRQAPGKGLEWVAFISYDGSNDYYADSVKGRFTISRDNS KNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS; or the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNLFSGSKSGNTASL TISGLQAEDEADYYCSSYTSSSTLNWGCGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFSFTMFGLHWVRQAPGKGLEWVAFISYDGSNDYYAHSVKGRFTISRDNS KNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS.

[0008] In embodiments, the fragment of the antibodies or antigen-binding fragments or variants thereof can include an F(ab), an Fv, or an scFv. In embodiments, the fragment of the antibodies or antigen-binding fragments or variants thereof can include a VhH.

[0009] Aspects are directed to a therapeutic antibody that binds to Vascular Endothelial Growth Factor (VEGF) protein comprising a variable domain and a constant domain. In embodiments, the constant domain is IgG and the variable domain comprises a framework region and a complementary determining means for binding to Vascular Endothelial Growth Factor (VEGF) protein. In embodiments, the constant region of the therapeutic antibody is IgG1 or IgG4. In embodiments, the therapeutic antibody can include any one of the antibodies according to Table 1 – Table 4.

[0010] Aspects are directed to a pharmaceutical composition that can include any of the antibodies described herein, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0011] Aspects are directed to methods of treating cancer in a subject that can include administering to a subject in need thereof an effective amount of any of the antibodies described herein, or the pharmaceutical composition described herein.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0012] Aspects are directed to the use of any of the antibodies described herein, or the pharmaceutical composition described herein for treating cancer. In some embodiments, any of the antibodies described herein, or any of the pharmaceutical compositions described herein can be use in treating cancer. In embodiments, the cancer comprises Ewing carcinoma, renal cell carcinoma, or lung adenocarcinoma.

[0013] Aspects of the invention are directed to a method of generating and / or increasing angiogenesis in a cancer in a subject that can include administering to the subject in need thereof an effective amount of the antibodies as described herein, or the pharmaceutical composition described herein.

[0014] Aspects of the invention are direct to nucleic acids encoding the antibodies as described herein.

[0015] Aspects are directed to vector comprising the nucleic acid as described herein.

[0016] Still further, aspects are drawn towards a cell comprising nucleic acids or vectors described herein.

[0017] Aspects are drawn to a cell producing the monoclonal antibody as described herein. BRIEF DESCRIPTION OF THE FIGURES

[0018] FIG. 1 shows VEGF family overview. The VEGF family ligands and receptors. Bevacizumab blocks the VEGF-ANEGFR-2 pathway by binding to all isoforms of VEGF-A. (A) An overview of the VEGF gene which contains eight exons. Exon splicing results in the production of different VEGF isoforms. TGA - stop codon, ATG - start codon. Image from Ladomery, M. R., Harper, S. J., & Bates, D. 0. (2007). Alternative splicing in angiogenesis: The vascular endothelial growth factor paradigm. Cancer Letters. (B)

[0019] FIG. 2 shows quantification of secreted VEGF-A165. Assessment of VEGF-A165 secretion from cell lines in the lab using ELISA Max Deluxe kit. Supernatants were gathered from each well over four days. The media was not replaced, and cells were not passaged to mimic a stressed state. RMPI and DMEM media were used as control conditions. TC32 refers to a Ewing sarcoma cell line; SKRC++ refers to a human renal carcinoma cell line; CADO refers to a human Ewing sarcoma cell line; A549 refers to a human lung adenocarcincoma cell line.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0020] FIG. 3 shows overview of antibody discovery. Schematic diagram demonstrating the steps in the antibody discovery process. (A) Flowchart depicting antibody removal from candidate pool. Antibodies grouped based on VH germline family and competition binding results. (B)

[0021] FIG. 4 shows competition binding of antibody candidates. Percent blockage was determined by comparison to PBST. Lighter colors represent inhibition and dark blue colors represent minimal inhibition. Candidate E5-D4 demonstrated simultaneous binding to Bevacizumab and VEGF-A165. The experiment was performed with two plates labeled set one (A) and set two (B) which were performed using the same conditions.

[0022] FIG. 5 shows germline sequencing of antibody candidates. The germline sequencing for the eight antibody candidates that were included in the in vitro functional assays. The FR, CDR, and gene names were identified using the IMGT database. Letters in red represent deviations from the expected germline sequence and represent somatic mutations. The numbers in parenthesis represent replicates of our candidates identified during phage panning.

[0023] FIG. 6 shows germline sequencing of antibody candidates (continued). The germline sequencing for the eight antibody candidates that were included in the in vitro functional assays. The FR, CDR, and gene names were identified using the IMGT database. Letters in red represent deviations from the expected germline sequence and represent somatic mutations. The numbers in parenthesis represent replicates of our candidates identified during phage panning.

[0024] FIG. 7 shows kinetic characterization of anti-VEGF-A antibodies. Antibodies were diluted in PBS to concentrations of 25 nM, 12.5 nM, and 6.25 nM. The antibodies were loaded for 30 seconds, association occurred for 60 seconds, and dissociation occurred for 300 seconds. The regeneration buffer was 0.l M Glycine-HCl and the neutralization buffer was PBS-T.

[0025] FIG. 8 shows kinetic characterization of anti-VEGF-A antibodies. Antibodies were diluted in PBS to concentrations of 25 nM, 12.5 nM, and 6.25 nM. The antibodies were loaded for 30 seconds, association occurred for 60 seconds, and dissociation occurred for 300 seconds. The regeneration buffer was 0.1 M Glycine-HCl and the neutralization buffer was PBS-T. (A) Results of kinetic characterization of anti-VEGF-A antibody candidates. E5-F3 and ES-Al obtained Koff rates that were below the detection level for the OctetRed96. (B)Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0026] FIG. 9 shows HUVEC proliferation assay. The effects of antibody candidates, bevacizumab, and irrelevant antibody control on VEGF-induced HUVEC proliferation. An irrelevant anti-SARS-2 antibody that does not regulate angiogenesis was used as a control for comparison. The experiment was performed with two plates labeled set one (A) and set two (B).

[0027] FIG. 10 shows dose-dependent inhibition of HUVEC proliferation. Bevacizumab and E5-F3 inhibit HUVEC proliferation in a dose-dependent manner. The mean value of the VEGF-A only group was established as the reference point for 100% VEGF-A165 proliferation. The mean value of the No VEGF-A group was established as the reference for 0% VEGF-A65 proliferation.

[0028] FIG. 11 shows binding affinity of E5-F3 to VEGF-A165, VEGF-A121, VEGF-C and VEGF-D. The binding affinity of Bevacizumab and E5-F3 to VEGF-A165 and VEGF-A121 was assessed via ELISA. Each sample was tested in triplicate. Results demonstrated the binding of E5-F3 and Bevacizumab to VEGF-A121 and VEGF-A165. E5-F3 and Bevacizumab did not bind to VEGF-C and VEGF-D.

[0029] FIG. 12 shows HUVEC Tube Formation Assay. HUVECs were seeded on Matrigel- coated plates at 4 x 105 cells / mL and incubated with experimental conditions for approximately 16 hours. (A) The images were photographed at a scale of 5x and 1Ox magnification and analyzed using the Angiogenesis plug-in for Image J software. The number of nodes and number of junctions (B) were used to quantify angiogenesis. Both E5-F3 and Bevacizumab were able to inhibit HUVEC tube formation. (B) Collaborated with Laura Miguens on assay. DETAILED DESCRIPTION

[0030] Aspects described herein can lead to the inhibition of the VEGF-A / VEGFR-2 pathway, thereby restructuring abnormal tumor vasculature into a normalized vasculature. In embodiments, anti-VEGF-A antibodies have been discovered which were effective in inhibiting angiogenesis. Without wishing to be bound by theory, we developed and validated monoclonal antibodies against VEGF-A for inhibiting angiogenesis and / or restructuring abnormal tumor vasculature.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0031] The present disclosure provides antibodies or antigen-binding fragments or variants thereof that specifically binds Vascular Endothelial Growth Factor (VEGF) protein.

[0032] The present disclosure further provides therapeutic antibodies or antigen-binding fragments or variants thereof that specifically binds Vascular Endothelial Growth Factor (VEGF) protein.

[0033] Still further, the present disclosure provides a method of treating cancer by administering to a subject in need thereof an effective amount of the anti-VEGF antibody or anti-VEGF therapeutic antibody.

[0034] Also, the present disclosure provides a method of inhibiting angiogenesis in a subject by administering to the subject an effective amount of the anti-VEGF antibody or anti-VEGF therapeutic antibody.

[0035] Also, the present disclosure provides a method of inhibiting VEGF-induced proliferation of endothelial cells in a subject by administering to the subject an effective amount of the anti-VEGF antibody or anti-VEGF therapeutic antibody.

[0036] Also, the present disclosure provides a method of restructuring abnormal tumor vasculature into a normalized vasculature by administering to the subject an effective amount of the anti-VEGF antibody or anti-VEGF therapeutic antibody.

[0037] Further, the present disclosure is drawn to a nucleic acid encoding an anti-VEGF mAb.

[0038] The present disclosure also provides an antibody of the present disclosure for use in therapy. More particularly, the present disclosure provides an antibody of the present disclosure for use in treatment of cancer. Further, the present disclosure provides the use of an antibody of the present disclosure in the manufacture of a medicament for the treatment of cancer.

[0039] Disclosed herein are isolated, recombinant monoclonal antibodies that specifically bind to VEGF.

[0040] Herein, "specifically binds" or "immunoreacts with" can refer to the antibody reacting with one or more antigenic determinants of VEGF and does not react with other polypeptides.

[0041] The term “isolated” as used herein, such as with respect to cells, proteins or polypeptides, or nucleic acids (e.g., DNA or RNA), can refer to those cells, proteins or polypeptides, or nucleic acids that are purified to some degree from endogenous materials TheDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 term “isolated” can also refer to a nucleic acid or peptide that is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. For example, an “isolated nucleic acid” can include nucleic acid fragments which are not naturally occurring as fragments and would not be found in the natural state. “Isolated” can also refer to cells or polypeptides which are isolated from other cellular proteins or tissues. Isolated polypeptides can include both purified and recombinant polypeptides.

[0042] The term “recombinant” as it pertains to polypeptides (such as antibodies) or polynucleotides can refer to a form of the polypeptide or polynucleotide that does not exist naturally, a non-limiting example of which can be created by combining polynucleotides or polypeptides that would not normally occur together.

[0043] The VEGF antibodies described herein bind to VEGF. In one embodiment, the VEGF antibodies have high affinity and high specificity for VEGF. In some embodiments, the VEGF antibodies can block the VEGF-A / VEGFR-2 axis, such as by blocking VEGF-A from binding to VEGFR-2. In some embodiments, blocking of VEGF-A binding to VEGFR-2 will restore the abnormal tumor vasculature and promote normal vasculature. Without wishing to be bound by theory, the antibodies described herein can counteract the effects of VEGF-A induced angiogenesis, and therefore prevent and / or restore normal vasculature.

[0044] Herein, an "antibody" can refer to an immunoglobulin molecule comprising two heavy chains (HCs) and two light chains (LCs) interconnected by disulfide bonds. The amino terminal portion of each LC and HC includes a variable region of about 100-120 amino acids primarily responsible for antigen recognition via the complementary determining region (CDRs) contained therein. The CDRs are interspersed with regions that are well-known and generally conserved among and between species (e.g., mouse and human), which are termed framework regions (FRs). The amino acids comprising the CDRs and the framework regions, respectively, can be readily identified for a heavy or light chain variable region by one of ordinary skill in the art, since they have been previously defined

[0045] In embodiments, the CDRs are interspersed with FRs. Antibodies disclosed herein have four FRs, termed FR1, FR2, FR3, and FR4. In embodiments, the FRs are human FRs (e.g., Antibody Engineering: Methods and Protocols (Damien Nevoltris and Patrick Chames eds., 3d ed.2018)).Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0046] The three CDRs of the light chain (LC) are referred to as "LCDR1, LCDR2, and LCDR3," and the three CDRs of the heavy chain (HC) are referred to as "HCDR1, HCDR2, and HCDR3." The functional ability of an antibody to bind a particular antigen is largely determined by the six CDRs. Assignment of amino acids to CDR domains within the LCVR and HCVR regions of the antibodies of the present disclosure is based on the IGMT numbering conventions.

[0047] The constant region of the antibody defines the isotype of an antibody. The antibodies of the present disclosure include IgG. IgG antibodies can be further divided into subclasses, e.g., IgG1, IgG2, IgG3, IgG4. In a particular embodiment, the antibodies of the present disclosure are IgG1. The carboxy-terminal portion of each HC defines a constant region primarily responsible for effector function. In a particular embodiment, the antibodies of the present disclosure have one or more modifications in the constant region of each HC that reduces effector function.

[0048] The tumor microenvironment (TME) consists of pro-angiogenic growth factors, such as vascular endothelial growth factor (VEGF), that contribute to the abnormal vasculature of tumor cells. The VEGF family consists of growth factors such as VEGF-A, VEGF-B, VEGF- C, VEGFD, and placental growth factors. Of the growth factors, VEGF-A is the major ligand as it regulates angiogenesis and stimulates the migration of endothelial cells.

[0049] The VEGF family is led by VEGF-A, a signaling protein responsible for blood vessel formation, and its receptor VEGFR-2. The VEGF-A / VEGFR-2 axis activates angiogenesis and promotes tumor development. The upregulation of VEGF-A produces a structurally and functionally abnormal tumor vasculature with features such as irregular branching patterns. Therefore, VEGF is a target for immunotherapy.

[0050] VEGF-A has isoforms that are produced via alternate mRNA splicing, such as VEGF-Al65 and VEGF-Al21 (Holmes & Zachary, 2005). VEGF-A165 is an isoform of particular interest because VEGF-Al65 is the most abundant in the human body and has pro- angiogenic properties (Sedlar et al., 2021).

[0051] As used herein, the term “VEGF antibody” can refer to an antibody that binds VEGF-A and, without wishing to be bound by theory, disrupts VEGF from binding to VEGFR- 2. In embodiments, antibodies described herein can bind to VEGF-A165, VEGF-A121, or both VEGF-A165 and VEGF-A121. Referring to FIG 11, for example, antibodies as describedDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 herein are able to bind both VEGF-A121 and VEGF-A165 indicating broader binding specificity.

[0052] Antibodies within the scope of the antibody are the disclosed antibodies and functional equivalents thereto. Functional equivalent antibodies comprise different specific amino acid residues but maintain binding activity to VEGF. Functional equivalent antibodies would differ insubstantially in their abilities to disrupt VEGF from binding to VEGFR-2, for example, and therefore have a therapeutic effect.

[0053] The amino acid sequences of the monoclonal VEGF antibodies are provided below: Table 1. Amino acid sequences of heavy chain variable region (HCVR). T5-H12QVQLVQSGGALVRPGGSLRLSCVASGFTFNDYEMNWVRQAPGTGLEWVSSVTARGGTEYYADSVKGRFTISRDNSKNTVYLQMNSLRGEDTAVYYCARSARDSYGGDYFDYWGQGTLV TVSS I V R S R S R S R S R V R V R V R V D V Y V N VDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 E5-A3QVQLVQSGGGLVPGRSLRLSCAASGFTFDDYAVHWVRQAPGKGLEWVSGLSWNSGSVGYADSVRGRFTISRDNAGNSLYLQMNGLRAEDTAFYYCAKGTKDSGASGPYFFDY WGQGTLVTVSS GGV W T R S L V NYS Y S Y S T S SHV H V K V K S K S Y T Y TTable 2. Amino acid sequences of light chain variable region (LCVR).Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 T5-H12SYELTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDHPVFGGGTKLAVL E5-G11QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPAQAPVLVVYDDSDRPSGIPERFFIFPPPPLRRRRDRRPP PPPADDDDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 E5-G12QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKSGNTASLTISGLQAEDEADYFCSSYTITQHRGFGGGTKLTVP E5-E6QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSSS S(CDRs). Antibody name CDR1 CDR2 CDR3 T5-H12GFTFNDYE VTARGGTE ARSARDSYGGDYFDYDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 E5-G5GFIFNNYW IERYGGEI ARPSWNSGSYFDYT5-D12GFTFSDYW IERYGAEE ARPTFNSGSYFDYTable 4. Amino acid sequences of light chain (LC) complementarity determining regions (CDRs). Antibody name CDR1 CDR2 CDR3 T5-H12NIGSKS YDS QVWDSSSDHPVDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 E5-D8SSDLGGHNF DVF SSYTITSIVVT5-F10SSDLGGHNF DVF SSYTITSIVV[ ] ur er sc ose ere n are an o y ragmens, suc as we -c arac er ze a s (e.g., Fab, Fab', F(ab')2, F(ab)2,), Fvs (the variable region of the light chain and the variable region of the heavy chain expressed as two chains), and single-chain fragments (e.g., single- chain variable region fragments, scFv, scFv-Fc, and single chain Fabs, scFab), which also bind to VEGF. Antibody fragments can include aptamers, minibodies, and diabodies. Methods of making these fragments are routine (see, e.g., Antibody Engineering: Methods and Protocols (Damien Nevoltris and Patrick Chames eds., 3d ed.2018)).

[0055] The antibodies and fragments thereof disclosed herein can also include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized, or chimeric antibodies.

[0056] The phrase “complementary determining means” as used herein describes the six complimentary determining regions (CDRs) that collectively form specific interactions with EDIL3. CDRs within the scope of complementary determining means are the disclosed CDRs and functional equivalents thereto. Functional equivalent CDRs comprise different specific amino acid residues but maintain binding to VEGF. Functional equivalent CDRs would differ insubstantially in their ability to disrupt VEGF from binding to its target, for example, and therefore have a therapeutic effect.

[0057] Contemplated herein are conservative variants of the disclosed antibodies and fragments thereof. A protein is a conservative variant where it contains conservative amino acid substitutions that do not substantially affect or decrease the affinity of a protein. For example, an antibody that binds VEGF can include at least 1, 2, 5, 10, or 15 conservative substitutions, for example, in a constant domain, and still bind VEGF. Conservative amino acid substitution tables providing functionally similar amino acids are well-known to one of ordinary skill in the art. The following groups are examples of amino acids that are considered conservative substitutions for one another: 1) serine (S), threonine (T); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucineDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 (I), leucine (L), methionine (M), valine (V); and 6) phenylalanine (F), tyrosine (Y), tryptophan (W).

[0058] Herein, a "degenerate variant" can refer to a polynucleotide encoding a polypeptide (such as an antibody or fragment thereof) that includes a sequence that is degenerate based on the genetic code (i.e., the 20 natural amino acids can be specified by more than one codon). All degenerate nucleotide sequences encoding the disclosed antibody and fragment polypeptide sequences are included.

[0059] Further contemplated are variants of the disclosed antibodies and fragments thereof with a sequence identity of at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the sequences according to Table 1A to Table 4B. Herein, “sequence identity“, “sequence homology”, or “sequence similarity” can refer to the similarity between amino acid or nucleic acid sequences, which is expressed as the similarity between the sequences. Sequence identity can be frequently measured as percent identity, in which two sequences are considered more similar the higher the percentage. Homologs or variants of a polypeptide or nucleic acid molecule possess a relatively high degree of sequence identity when aligned using standard methods, which are well-known. Ceslovas Venclovas, Methods for Sequence-Structure Alignment in Homology Modeling: Methods and Protocols, 55-82 (Andrew Orry and Ruben Abagyan, eds., 2012)).

[0060] Herein, “binding” (or “binds”) can refer to the well understood interaction between and antibody and a target protein, peptide, or polysaccharide. Binding can be measured in a variety of ways (see, e.g., Antibody Engineering: Methods and Protocols (Damien Nevoltris and Patrick Chames eds., 3d ed. 2018)). In embodiments, binding (e.g., binding activity) is measured by an ELISA assay. See Example 2, for example. A particular antibody or protein binds to a particular target protein, peptide, or polysaccharide and does not bind in a significant amount to other proteins or polysaccharides present in a sample or subject disclosed herein. Binding occurs between the disclosed antibodies and fragments thereof and an epitope of VEGF. Herein, "epitope" can refer to discrete sites of an antigen recognized by the disclosed antibodies and fragments thereof. Epitopes may be linear or three-dimensional.

[0061] The strength, or affinity of immunological binding interactions can be expressed in terms of the equilibrium binding constant (Kd) of the interaction, wherein a smaller Kdrepresents a greater affinity. An antibody binds to a target protein when the interaction has aDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 KD of less than 10-6molar, such as less than 10-7molar, less than 10-8molar, less than 10-9molar, or less than 10-10molar.

[0062] Functionally, the binding affinity of the anti-VEGF antibody is within the range of 10−5M to 10−12M. For example, the binding affinity of the anti-VEGF antibody is from 10−6M to 10−12M, from 10−7M to 10−12M, from 10−8M to 10−12M, from 10−9M to 10−12M, from 10−5M to 10−11M, from 10−6M to 10−11M, from 10−7M to 10−11M, from 10−8M to 10−11M, from 10−9M to 10−11M, from 10−10M to 10−11M, from 10−5M to 10−10M, from 10−6M to 10−10M, from 10−7M to 10−10M, from 10−8M to 10−10M, from 10−9M to 10−10M, from 10−5M to 10−9M, from 10−6M to 10−9M, from 10−7M to 10−9M, from 10−8M to 10−9M, from 10−5M to 10−8M, from 10−6M to 10−8M, from 10−7M to 10−8M, from 10−5M to 10−7M, from 10−6M to 10−7M, or from 10−5M to 10−6M.

[0063] Referring to FIG. 4, results demonstrate the anti-VEGF antibodies competes with Bevacizumab for binding to biotinylated VEGF-A165, indicating a shared epitope VEGF. Accordingly, in certain embodiments, the anti-VEGF antibody binds an epitope that overlaps with that of Bevacizumab or binds to an epitope that is the same as or substantially similar to that of Bevacizumab. Accordingly, aspects of the invention are drawn to an anti-VEGF antibody of the present disclosure, wherein the anti-VEGF antibody competes with binding to VEGF or binds the same epitope on VEGF as Bevacizumab.

[0064] A first binding protein (e.g., antibody or ligand) “binds to the same epitope” as a second binding protein (e.g., antibody or ligand) if the first binding protein binds to the same site on a target antigen that the second binding protein binds, or binds to a site that overlaps (e.g., 50%, 60%, 70%, 80%, 90%, or 100% overlap, e.g., in terms of amino acid sequence or other molecular feature (e.g., glycosyl group, phosphate group, or sulfate group)) with the site that the second binding protein binds.

[0065] A first binding protein (e.g., antibody or ligand) “competes for binding” with a second binding protein (e.g., antibody) if the binding of the first binding protein to its epitope decreases (e.g., by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or more) the amount of the second binding protein that binds to its epitope. The competition can be direct (e.g., the first binding protein binds to an epitope that is the same as, or overlaps with, the epitope bound by the second binding protein), or indirect (e.g., the binding of the first bindingDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 protein to its epitope causes a steric change in the target compound that decreases the ability of the second binding protein to bind to its epitope).

[0066] Those skilled in the art will recognize that it is possible to determine, without undue experimentation, if a monoclonal antibody has the same specificity as a monoclonal antibody of the invention by ascertaining whether the former prevents the latter from binding to VEGF. For example, if the monoclonal antibody being tested competes with the monoclonal antibody of the invention, as shown by a decrease in binding to VEGF by the monoclonal antibody of the invention, then the two monoclonal antibodies bind to the same, or to a closely related, epitope.

[0067] Another way to determine whether a monoclonal antibody has the specificity of a monoclonal antibody of the invention is to pre-incubate the monoclonal antibody of the invention with the VEGF protein, with which it is normally reactive, and then add the monoclonal antibody being tested to determine if the monoclonal antibody being tested is inhibited in its ability to bind VEGF. If the monoclonal antibody being tested is inhibited then, in all likelihood, it has the same, or functionally equivalent, epitopic specificity as the monoclonal antibody of the invention. Screening of monoclonal antibodies of the invention can be also carried out by utilizing VEGF and determining whether the test monoclonal antibody is able to neutralize VEGF.

[0068] The antibodies herein are monoclonal antibodies ("mAbs"). Various procedures known within the art can be used for the production of monoclonal antibodies directed against a protein of the invention, or against derivatives, fragments, analogs homologs or orthologs thereof. mAbs can be produced, for example, by hybridoma technologies, recombinant technologies, phage display technologies, synthetic technologies (e.g., CDR or specificity- determining residue, SDR, grafting), or combinations of such or other technologies known in the art. mAbs are antibodies derived from a single copy or clone including, for example, any eukaryotic, prokaryotic or phage clone. In some embodiments, VEGF antibodies can be generated using Single B cell cloning technology. A variety of well-known methods and tools can be used for producing and purifying the mAbs disclosed herein, including vectors, for example, plasmids, virus, or other vehicles for polynucleotide insertion or expression, and hosts, for example, microbial, yeast, insect, and mammalian organisms (see, e.g., Process Scale Purification of Antibodies (Uwe Gottschalk, ed., 2d ed. 2017)). For example, a vector forDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 producing and purifying mAbs disclosed herein can contain a DNA segment encoding the monoclonal antibody described herein. In embodiments, the vector can be an adeno-associated virus (AAV), a retrovirus, a lentivirus vector, or the like.

[0069] Further antibodies having a complementary binding means can be prepared and screened by well-known methods, such as hybridoma, transgenic animals, and phage or yeast display (see, e.g., Monoclonal Antibodies: Methods and Protocols (Vincent Ossipow and Nicolas Fischer, eds., 2d ed. 2014)). Antibodies having equivalent complementary binding means differ in their amino acid sequence but perform the same function of binding the target through CDR-target interaction acting as (inhibitor / agonist / antagonist) to achieve the same result (inhibiting tumor growth). Preferably, the complementary binding means functions through the same epitope as the disclosed antibodies.

[0070] In embodiments, an antibody can comprise an Fc variant comprising an amino acid substitution which alters the antigen-independent effector functions of the antibody, in particular the circulating half-life of the antibody.

[0071] In embodiments, antibodies disclosed herein can comprise an Fc variant that can have reduced or eliminated glycosylation (e.g., N- or O-linked glycosylation).

[0072] In embodiments, antibodies disclosed herein can comprise an Fc variant that can either increase or decrease binding to FcRn when compared to antibodies lacking these substitutions, therefore, have an increased or decreased half-life in serum, respectively.

[0073] In embodiments, antibodies disclosed herein can comprise an Fc variant comprising mutations introduced to the constant regions of the mAb such that the antibody dependent cell- mediated cytotoxicity (ADCC) activity of the mAb is altered. For example, the mutation is a LALA mutation in the CH2 domain which reduces ADCC activity.

[0074] The antibodies and fragments thereof disclosed herein can be used in therapy. In embodiments, the antibodies and fragments thereof disclosed herein can be used to treat, prevent (such as through prophylactic treatment), or ameliorate a cancer or metastasis. In embodiments, the cancer can comprise Ewing sarcoma, renal cell carcinoma, or lung adenocarcinoma. Non-limiting examples of other cancers that can be treated include melanoma, non-small-cell lung cancer (NSCLC), small cell lung cancer (SCLC), chronic lymphocytic leukemia (CLL; such as B cell CLL or T cell CLL), classical Hodgkin lymphoma (cHL), head and neck squamous cell carcinoma (HNSCC), colorectal cancer (CRC), gastricDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 cancer, hepatocellular carcinoma (HCC), primary mediastinal large B-cell lymphoma (PMLBCL), bladder cancer, urothelial cancer, endometrial cancer, cervical cancer, breast cancer (e.g., triple negative breast cancer), Merkel cell carcinoma (MCC), and microsatellite instability high (MSI-H) or DNA mismatch repair deficient (dMMR) adult and pediatric solid tumors.

[0075] In some embodiments, the method for treating cancer can include the use of a pharmaceutically effective amount of one or more VEGF antibodies that are administered to a subject with cancer. In some embodiments, the method of treating cancer leads to the prevention of angiogenesis and / or the restoration of normal vasculature. In some embodiments, the method of treating cancer can counteract the effects of the VEGF-A / VEGFR-2 axis on angiogenesis activation and tumor development.

[0076] In some embodiments, the method for treating cancer can include one or more additional therapies to treat cancer. In some embodiments, the additional therapy to treat cancer is a therapy that can modify angiogenesis. In some embodiments, the additional therapy to treat cancer is a therapy that can modify checkpoint blockade.

[0077] Herein, "preventing" a disease can refer to inhibiting the full development of a disease, such as cancer. "Treating" can refer to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop, such as a reduction in tumor burden or a decrease in the number of size of metastases. "Ameliorating" can refer to the reduction in the number or severity of signs or symptoms of a disease, such as cancer. A "prophylactic" treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing a pathology, such as cancer. The antibodies and fragments thereof disclosed herein can be administered to subjects or patients. Herein, “administration” can refer to the act of the attending physician or caregiver, prescribing the agent for administration and thereby causing the application of an agent to a subject, through ingestion, infusion, injection, or any other means, whether self-administered or administered by a clinician or other qualified care giver. For example, administration of the mAb disclosed herein can abrogate or inhibit or interfere with an activity of the VEGF protein. The term “therapeutic” in conjunction with antibody disclosed herein refers to an antibody suitable for use in human treatment of cancer. In embodiments, such an antibody can have aDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 KD of less than 10-6molar, such as less than 10-7molar, less than 10-8molar, less than 10-9molar, or less than 10-10molar and any toxic or detrimental effects of the antibody are outweighed by the therapeutic beneficial effects.

[0078] Herein, a “subject” includes both human patient and veterinary subjects, including human and non-human mammals. In embodiments, the subject or patient has or has a risk of cancer.

[0079] A pharmaceutical composition of the present disclosure contains an "effective" or "therapeutically effective" amount, as used interchangeably herein, of a monoclonal antibody of the present disclosure. The dosages and dosage regimen to achieve the desired therapeutic result depending on the means of administration and may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the monoclonal antibody to elicit a desired response in the individual. The dosage administered to a subject (e.g., a patient) of the antigen-binding polypeptides described herein is typically 0.1 mg / kg to 100 mg / kg of the patient's body weight, between 0.1 mg / kg and 20 mg / kg of the patient's body weight, or 1 mg / kg to 10 mg / kg of the patient's body weight. An effective amount is also one in which any toxic or detrimental effects of the monoclonal antibody of the present disclosure are outweighed by the therapeutically beneficial effects.

[0080] A pharmaceutical composition of the present disclosure can be formulated to be compatible with its intended route of administration. Non-limiting examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration.

[0081] The pharmaceutically acceptable carriers of use are conventional (e.g., as described in Remington, The Science and Practice of Pharmacy, 22nd Edition, Loyd V., ed., Pharmaceutical Press, 2012). In general, the nature of the carrier will depend on the mode of administration. For instance, parenteral formulations typically comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids, such as water, physiological saline, balanced salt solutions or the like as a vehicle. Pharmaceutical compositions can additionally include minor amounts of non-toxic auxiliary substances for stability.

[0082] In embodiments, the carrier can be sterile and / or suspended or otherwise contained in a unit dosage form including one or more measured doses of the composition suitable for administration to a subject of an effective amount of the antibodies and fragments thereofDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 disclosed herein. Medications for use in treatment can also be included in embodiments. In embodiments, the unit dosage form can be in a sealed vial that contains sterile contents or a syringe for injection into a subject, lyophilized for subsequent solubilization and administration, or in a solid or controlled release dosage.

[0083] Herein, an “effective amount” can refer to a quantity sufficient to achieve a desired effect in a subject. For instance, this can be the amount necessary to prevent, treat, or ameliorate a disease, for example, inhibiting or suppressing cancer. In embodiments, an effective amount is the amount necessary to eliminate, reduce the size, or prevent metastasis of cancer or a tumor. Efficacy is first evident in the cellular response, for which a variety of in vitro and cell assays are well-known to measure. Kristina V. Kitaeva et al., Cell Culture Based In vitro Test Systems for Anticancer Drug Screening, 8 Front. Bioeng. Biotechnol.322(2020)). In embodiments, an effective amount is the amount necessary to significantly inhibit or reduce cancer cell proliferation or migration, invasion, or adhesion. A cellular response manifests as significantly reduced tumor size, reduced or inhibited disease progression, and improvement in survival in a subject. More particularly, an effective amount provides improvement in important cancer endpoints, Overall Survival (OS), Disease-Free Survival (DFS), Objective Response Rate, Complete Response Rate or Progression Free Survival (PFS). See Dept. of Health and Human Services, Food and Drug Admin, Clinical Trial Endpoints for the Approval of Cancer Drugs and Biologics: Guidance for Industry (2018); E.A. Eisenhauer et al., New Response Evaluation Criteria in Solid Tumours: Revised RECIST Guideline (Version 1.1), 45 Eur. J. Cancer 228 (2009).

[0084] The mAbs disclosed herein can be used in conventional methods relating to the localization and / or quantitation of an VEGF protein (e.g., for use in measuring levels of the VEGF protein within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like).

[0085] The mAbs disclosed herein can be used in methods relating to the isolation of an VEGF polypeptide by conventional techniques, such as immunoaffinity, chromatography or immunoprecipitation. Antibodies directed against an VEGF protein (or a fragment thereof) can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0086] Aspects are drawn towards a method of identifying a subject in need of an anti- angiogenesis therapy. For example, embodiments can comprise detecting VEGF expression or protein levels in a sample obtained from a subject. “Detecting” can refer to quantitative, semi- quantitative, qualitative, or other methods for determining an analyte, e.g., VEGF, in a sample. For example, detecting can comprise conventional methods, such as immunohistochemistry of the tumor and / or the tumor microenvironment; measuring levels of VEGF, such as circulating VEGF; and / or measuring levels of VEGF antibody. In embodiments, the elevated levels of VEGF expression or protein can be indicative of a subject in need of an anti-angiogenesis therapy.

[0087] In embodiments, the VEGF expression or protein levels can be compared to a control sample and, if changed (e.g., above or below) when compared to the control sample, the subject can be identified as a subject in need of an anti-angiogenesis therapy. As used herein, “changed as compared to a control” sample or subject is understood as having a level of the analyte or diagnostic or therapeutic indicator (e.g., marker, such as VEGF) to be detected at a level that is statistically different than a sample from a normal, untreated, or abnormal state control sample. Determination of statistical significance is within the ability of those skilled in the art, e.g., the number of standard deviations from the mean that constitute a positive or negative result.

[0088] Aspects are drawn towards methods of preventing, reducing or inhibiting angiogenesis. “Angiogenesis” can refer to the formation and spread of blood vessels. For example, “angiogenesis” can refer to a process involving tissue angiogenesis, such as, the proliferation, migration and invasion of vascular endothelial cells and the development of new capillaries. Referring to FIG. 9, anti-VEGF antibodies as described herein inhibited proliferation of human umbilical vein endothelial cells, a cell line commonly used to measure angiogenesis.

[0089] The mAbs of the present disclosure can also be used in combination therapy. In embodiments, the subject is treated with an mAb disclosed herein in combination with one or more additional therapies to treat cancer, for example, radiation, surgery, bone marrow transplantation, chemotherapy, immunotherapy, hormone therapy, or targeted therapy. Use of the mAbs of the present disclosure in combination with chemotherapy or immunotherapy is preferred. Most preferably, the additional treatment is directed to angiogenesis and / orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 checkpoint blockade. Accordingly, aspects of the invention are drawn towards administering an mAb disclosed herein in combination with an anti-angiogenic agent and / or an immune checkpoint blockade therapy.

[0090] As used herein, “combination” therapy or use in combination refers to the administration of the mAbs of the present disclosure to a patient in conjunction with (i.e., before, simultaneously, or following) any number of relevant treatments. Such treatments include, but are not limited to, agents such as an angiogenesis inhibitor and / or an immune checkpoint inhibitor.

[0091] An “angiogenesis inhibitor” or “anti-angiogenic agent” can refer to a substance that inhibits the growth of new blood vessels. In embodiments, the angiogenesis inhibitor can comprise cytokines, an EDIL-3 antibody, a VEGF inhibitor, a tyrosine kinase inhibitor, or a combination thereof. For example, the VEGF inhibitor comprises a VEGF antibody. For example, the angiogenesis inhibitor comprises axitinib, bevacizumab, cabozantinib, everolimus, lenalidomide, Lenvatinib mesylate, pazopanib, ramucirumab, regorafenib, sofafenib, sunitinib, thalidomide, vandetanib, and ziv-aflibercept.

[0092] Without wishing to be bound by theory, administering an angiogenesis inhibitor can synergize a patient’s response to an immune blockade therapy.

[0093] The term “immune checkpoint inhibitor” or “immune checkpoint blockade therapy” can refer to any compound inhibiting the function of an immune inhibitory checkpoint protein. Inhibition includes reduction of function and full blockade. In embodiments, immune checkpoint inhibitors can be antibodies that specifically recognize an immune checkpoint protein. In embodiments, immune checkpoint inhibitors can include peptides, antibodies, nucleic acid molecules, and small molecules.

[0094] In embodiments, the checkpoint blockade therapy can comprise a CTLA4 antibody, a PD-L1 antibody, a PD-1 antibody, a LAG-3 antibody, or a combination thereof. For example, the CTLA4 antibody can comprise ipilimumab, tremelimumab, or a combination thereof. For example, the PD-L1 antibody can comprise atezolizumab, avelumab, duvalumab, or a combination thereof. For example, the PD-1 antibody can comprise pembrolizumab, nivolumab, cemiplimab, or a combination thereof. For example, the LAG-3 antibody can comprise relatlimab.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0095] Cellular therapies, such as chimeric antigen receptor (CAR) cell therapies, are also provided herein. For example, the cell can be a CAR T-cell (e.g., T lymphocyte, a CD4+ T cell, a CD8+ T cell, or the combination thereof) or a CAR NK-cell. CAR cell therapies redirect a patient’s T-cells and / or NK-cells to kill tumor cells by the exogenous expression of a CAR on a T-cell or NK-cell, for example. A CAR can be a membrane spanning fusion protein that links the antigen recognition domain of an antibody to the intracellular signaling domains of the T- cell receptor and co-receptor or NK-cell receptor.

[0096] In one embodiment, monospecific CAR cells are provided. For example, the VEGF antibodies described herein can be used as the targeting moiety for the CAR cell. The CAR cell can be an engineered cell comprising a chimeric antigen receptor, wherein the chimeric antigen receptor comprises an extracellular ligand binding domain that is specific for VEGF.

[0097] In another embodiment, the VEGF antibodies described herein can be used as a payload for armored CAR-cell therapies. A suitable cell can be used, for example, that can secrete a VEGF antibody of the invention (or alternatively engineered to express a VEGF antibody as described herein to be secreted). The anti-VEGF “payloads” to be secreted, can be, for example, minibodies, scFvs, IgG molecules, fusion molecules, and other antibody fragments as described herein. Upon contact or engineering, the cell described herein can then be introduced to a patient in need of a treatment by infusion therapies known to one of skill in the art.

[0098] In embodiments, CAR cells (i.e., CAR T cells or CAR NK cells) can be generated according to methods known in the art using lentivirus systems (via transduction), retrovirus systems (via transfection (electroporation)), and transposon systems (via PiggyBac). Useful for promoters for payloads that can be used in the generating of CAR-Ts include, for example, constitutive promoters (where the promoter is the same as for CAR-T, such as EF1a then IRES or 2A); inducible promoters (where the promoter is different from the promoter for CAR-T, such as NFAT, IL-2 prom); and genetically engineered promoters (such as a VEGF locus “knock in” of cytokine and / or a promoter that is under the control of an endogenous promoter).

[0099] The skilled artisan will recognize that CAR cells can be generated from cellular sources known to the skilled artisan. Non-limiting examples include T cells, NK cells, iPSC- derived cells (e.g., iPSC-derived T-cells and / or iPSC-derived NK cells), peripheral blood cells (e.g., peripheral blood mononuclear cells), cord blood cells, cell lines (e.g., NK92 cell line),Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 human embryonic stem cells (hESCs) and CD34+ hematopoietic progenitor cells (HPCs). See, for example, Lu, Hui, et al. "From CAR-T cells to CAR-NK cells: a developing immunotherapy method for hematological malignancies." Frontiers in Oncology (2021): 3151.

[0100] Equivalents

[0101] Various aspects of the present disclosure may be used alone, in combination, or in a variety of arrangements not specifically discussed in the embodiments described in the foregoing and is, therefore, not limited in its application to the details and arrangement of components set forth in the foregoing description or illustrated in the drawings. For example, aspects described in one embodiment may be combined in any manner with aspects described in other embodiments.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 EXAMPLES

[0102] Examples are provided below to facilitate a more complete understanding. The following examples illustrate the exemplary modes of making and practicing the invention. However, the scope is not limited to specific embodiments disclosed in these Examples, which are for purposes of illustration only, since alternative methods can be utilized to obtain similar results.

[0103]

[0104] EXAMPLE 1

[0105] Discovery of Vascular Endothelial Growth Factor (VEGF) Antibodies for a Normalized Tumor Vasculature

[0106] Abstract

[0107] Chimeric antigen receptor (CAR) T cells present a promising approach to cancer immunotherapy due to their longevity and functionality in the human body. Although CAR-T therapies have demonstrated success for blood cancers, further development is needed for solid tumor cancers. This discrepancy is partly due to the immunosuppressive tumor microenvironment (TME). The TME consists of pro-angiogenic growth factors, such as vascular endothelial growth factor (VEGF), that contribute to the abnormal vasculature of tumor cells. The VEGF family is led by VEGF-A, a signaling protein responsible for blood vessel formation, and its receptor VEGFR-2. The VEGF-A / VEGFR-2 axis activates angiogenesis and promotes tumor development. The up-regulation of VEGF-A produces a structurally and functionally abnormal tumor vasculature with features such as irregular branching patterns. The abnormal vasculature limits the egress of CAR-T cells out of the tumor vasculature and into the tumor tissue which reduces the efficiency of drug delivery. Without wishing to be bound by theory, the inhibition of the VEGF-A / VEGFR-2 pathway via the discovery of an anti-VEGF-A antibody will enhance immune cell trafficking of CAR-T cells that secrete anti-VEGF-A to the tumor sites and restore the abnormal tumor vasculature. In this project, a phage panning campaign was performed with the successful enrichment of anti- VEGF-A phage for further isolation and characterization of anti-VEGF-A antibodies. The antibody candidates were characterized in vitro via assays such as Bio-layer interferometry (BLI) and anti-angiogenic assays. The findings demonstrate that the lead antibody candidate exhibits anti-angiogenic properties in comparison to Bevacizumab, an FDA-approved anti-Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 VEGF-A antibody. The discovery of an anti-VEGF-A antibody for CAR-T cell therapy merits attention because it can help restore the abnormal tumor vasculature and improve cancer drug delivery for solid tumor cancers.

[0108] Introduction

[0109] Cancer is one of the leading causes of death worldwide and has affected the lives of many individuals. This disease arises from the uncontrollable proliferation and division of cells leading to the formation of malignant tumors. In recent years, groundbreaking advancements in cancer research have centered around immunotherapy – the use of a patient’s own immune system to attack unhealthy tumor cells. For example, autologous chimeric antigen receptor (CAR) T cell therapy involves the collection of T cells – or white cells that drive immune responses – from a patient’s bloodstream. On the other hand, allogeneic CAR-T cells involve the collection of T cells from a healthy, unrelated donor that are then infused into the patients that are receiving the therapy (Chen et al., 2023). Additionally, CAR-T therapy allows for the engineering of T cells that can recognize specific antigens – naturally expressed molecules found on the surface of cancer cells that can bind to T cells. The T cells are first engineered to express the engineered CAR that will then bind to the antigens on cancer cells. Then, the T cells are infused back into the patient’s body where they will become activated when they encounter the target antigen at the tumor site. The CAR-T cells will then undergo direct activation through the intracellular signaling domain of the CAR and secondary activation via cytokines, or small proteins involved in cell signaling (Zhang et al., 2017). Thus, the overall goal of CAR-T cell therapies is to target and kill tumor cells while also restoring the anti-tumor immunity at the tumor site.

[0110] The structure of a first-generation CAR-T cell includes an antigen-binding domain, an extracellular hinge, a transmembrane domain, and an intracellular signaling domain. The antigen-binding domain is located extracellularly and is responsible for recognizing the target antigen. The extracellular domain usually consists of a single-chain variable fragment (scFv) that can bind to surface antigens present on cancer cells. The scFv is a fusion protein comprised of the variable heavy (VH) and light chains (VL) of immunoglobulins (Ig) which are connected by a polypeptide linker. Next, the extracellular hinge connects the scFv to the transmembrane domain – a structure that serves to stabilize the CARs in the T cell membrane. Finally, the intracellular signaling domain is made up of an activation domain, such as a CD3ζ chain, toDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 allow for the activation of CAR-T cells after they bind to an antigen. In second-generation CAR-T cells, a co-stimulatory domain is added to allow for the long-term survival and proliferation of CAR-T cells in the body (Hong et al., 2020; Rafiq et al., 2020). Research studies showed that first-generation CAR-T cells have a short life span in vivo and CARs that only have the CD3ζ chain have demonstrated reduced T cell responses and proliferation. More specifically, initial clinical trials demonstrated minimal to negligible efficacy of first- generation CAR-T cells (Sterner & Sterner, 2021; Till et al., 2008). Co-stimulatory domains, such as CD28 and CD137, are thus able to improve the proliferation and cytotoxicity of CAR- T cells. For example, CD137 plays an important role in the response signal and survival of T cells and the memory of cytotoxic T cells. Third-generation CAR-T cells are made by combining multiple signaling domains with the goal of stronger increased cytokine production and enhanced persistence in the body (Zhang et al., 2017).

[0111] CAR-T cells can be designed in many ways to target specific factors. The immune system contains checkpoints that can send “off” signals and inhibit desired immune responses. Research studies have highlighted the importance of programmed cell death protein 1 (PD-1) and programmed death-ligand 1 (PD-L1) as immune checkpoints due to their ability to regulate T cell activity. PD-1 is a checkpoint protein on T cells that binds to its receptor PD-L1. As PD- 1 binds to PD-L1, inhibitory signals are sent to downregulate T cell activity and thus inhibit the killing of tumor cells (Gumber, 2022). The inhibitory motifs of PD-1 / PD-L1 can refer to two tyrosine motifs: an immune receptor tyrosine-based inhibitory motif (ITIM) and an immune receptor inhibitory tyrosine-based switch motif (ITSM). The binding of PD-1 to PD- L1 causes the phosphorylation of tyrosine residues in the PD-1 region of the ITSM domain which then leads to the inhibition of downstream signaling (Jiang et al., 2019). To combat this, CAR-T cells can be designed to secrete anti-PD-L1 antibodies that block the PD-1 / PD-L1 pathway and thus prevent T-cell exhaustion and inactivation (Suarez et al., 2016).

[0112] Currently, six CAR-T cell therapies have been approved by the Food and Drug Administration (FDA) for blood cancers such as myeloma and leukemia (Chen et al., 2023). However, efforts to design CAR-T cell therapies for solid tumor cancers have not yet achieved the same success. Obstacles in the development of CAR-T therapies for solid tumor cancers include factors such as the toxicity due to cytokine release and the role of the tumor microenvironment (TME). The deficit in FDA-approved treatments for solid tumors is due inDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 large part to the immunosuppressive TME. The TME is defined as the complex system surrounding tumor cells that provides the tumor with the necessary nutrients for tumor growth. For example, the TME consists of endothelial cells, blood, lymphatic vessels, and signaling molecules (Carmeliet & Jain, 2011; Finley & Popel, 2013). The TME not only serves as a physical barrier for CAR-T therapies but facilitates the recruitment of immunosuppressive T cells that dampen anti-tumor immune responses. Additionally, the presence of T cell inhibitory signals, such as PD-1, in the TME contributes to the dysfunction of CAR-T cell therapies (Rafiq et al., 2020). As previously mentioned, one method to overcome the T cell inhibitory signals is through the design of CAR-T cells that secrete anti-PD-L1 antibodies (Suarez et al., 2016).

[0113] The blood vessels surrounding solid tumors have structural irregularities, increased permeability, and are abnormal in function. This abnormal tumor vasculature is further characterized by leaky blood vessel networks and disorganized branching patterns. The increased permeability and leakiness of tumor blood vessels present a challenge as it allows for fluids and cancer cells to escape into surrounding tissues. Additionally, when tumor blood vessels are leaky this causes the leakage of fluid from blood vessels into the TME which can cause an increase in interstitial fluid pressure (Weis & Cheresh, 2005). Thus, the abnormal vasculature and increased interstitial fluid pressure present challenges for the delivery of cancer drugs as they can lead to poor penetration of cancer drugs through tumor vessels. Finally, the abnormal vasculature can hinder the extravasation of CAR T cells to the tumor site (Fukumura et al., 2018).

[0114] Angiogenesis and the VEGF family

[0115] Angiogenesis, the process by which blood vessels proliferate, is recognized as a hallmark of cancer. During processes of tumor cell growth and healing, angiogenesis facilitates the proliferation of blood vessels. This proliferation then allows for the transport of nutrients. Thus, targeting angiogenesis has emerged as a therapeutic strategy to inhibit the growth of solid cancer tumors. The abnormal tumor vasculature causes the tumor cells to undergo hypoxia – a condition marked by reduced oxygen supply to tumor cells (Fukumura et al., 2018). During hypoxia, tumor cells upregulate angiogenic factors as a method to enhance oxygen and nutrient supply. More specifically, higher levels of vascular endothelial growth factor (VEGF) are secreted by tumor cells when they experience hypoxia or other stress conditions (Ferrara, 2005). VEGF is a signaling protein that promotes the proliferation of endothelial cells to formDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 new blood vessels. Members of the VEGF family are commonly expressed in solid malignant tumors because the solid tumors are dependent on angiogenesis for blood vessel formation. Additionally, tumor cells need a steady blood supply that can provide oxygen and nutrients for their growth and survival.

[0116] The VEGF family consists of growth factors such as VEGF-A, VEGF-B, VEGF-C, VEGF-D, and placental growth factors. Of the growth factors, VEGF-A is the major ligand as it regulates angiogenesis and stimulates the migration of endothelial cells. VEGF-A has isoforms that are produced via alternate mRNA splicing, such as VEGF-A165 and VEGF-A121 (Holmes & Zachary, 2005). In many research studies, VEGF-A165 is selected as the isoform of interest. This preference is because VEGF-A165 is the most abundant in the human body and has pro-angiogenic properties (Sedlář et al., 2021). Additionally, responses to VEGF-A are mediated through the binding of VEGF-A to its receptor VEGFR-2. As VEGF-A binds to VEGFR-2 this causes endothelial cell migration, proliferation, and survival (FIG.1, panel A). VEGFR-2 is expressed on endothelial cells of blood and lymphatic vessels and is also the receptor for VEGF-C and VEGF-D (Goel et al., 2011; Jin et al., 2011). Thus, VEGF-A is an important target in cancer immunotherapy because of the significant role it plays in the upregulation of blood vessel formation.

[0117] The VEGF gene consists of eight exons with alternate splicing mechanisms that produce different pro-angiogenic or anti-angiogenic VEGF isoforms. The VEGF isoforms are generated by either 5’ splicing, 3’ splicing, or cassette exons (FIG.1, panel B). Cassette exons, also known as exon skipping, are created through alternate splicing. This process occurs when the exon of interest, positioned between two other exons in the sequence, is either included or excluded from the transcript. The cassette exons are marked by the inclusion of exon 7 which leads to the formation of VEGF-A165. On the other hand, the exclusion of exon 7 leads to the formation of VEGF-A121 (Ladomery et al., 2007).

[0118] Anti-angiogenic therapies have been designed as approaches to tackle solid tumor cancers and are designed to block the binding of VEGF-A to VEGFR-2. This is achieved through the design of anti-VEGF-A antibodies that bind to VEGF-A. For example, Bevacizumab (Avastin), the first anti-angiogenic antibody to be approved by the FDA in 2006, operates by binding to VEGF-A to block the VEGF-A / VEGFR-2 pathway. Bevacizumab was approved as a treatment for colorectal cancer and can bind to all isoforms of VEGF-A (Ferrara,Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 2005; Simó & Hernández, 2008). Another common method of anti-angiogenic therapy is the blockage of the VEGF-A / VEGFR-2 pathway with the development of an anti-VEGFR-2 antibody. Similar to an anti-VEGF-A antibody, an anti-VEGFR-2 antibody can bind to the VEGFR-2 receptor on the surface of endothelial cells and block the VEGF-A / VEGFR-2 pathway. For example, Ramucirumab is an FDA-approved monoclonal antibody that can bind to VEGFR-2 and is used for the treatment of non-small-cell lung cancer (NSCLC) (Cheng et al., 2023).

[0119] Research Objectives

[0120] Our aim of this study was to identify an anti-VEGF-A antibody that could restore the abnormal tumor vasculature through the inhibition of the VEGF-A / VEGFR-2 pathway. Without wishing to be bound by theory, an anti-VEGF-A165 antibody will produce a normalized tumor vasculature with enhanced immune cell trafficking to the tumor site and improved drug delivery.

[0121] We performed the steps of panning a 27-billion-member human scFv phage display library. A phage display library is a collection of bacteriophages presenting diverse proteins on their surface. This library allows for the screening of molecules that bind to specific antigens. Thus, the screening of large phage libraries allows for the identification of scFv antibodies against a target of interest (Alfaleh et al., 2020; Azzazy & Highsmith, 2002). Due to the small size (~25 kDa) of scFvs, the scFvs are reformatted into scFvs fused to the fragment crystallizable (Fc) to facilitate downstream experiments. The Fc region is approximately 50 kDa and plays a crucial role in immune responses by mediating effector functions that enhance the clearance of pathogens from the body (Schneider et al., 2021).

[0122] The results of the phage panning campaign demonstrated that there was successful enrichment of anti-VEGF-A165 phage. To discover an anti-VEGF-A antibody, we first performed a screening of individual phage colonies to identify the candidates that bind to soluble VEGF-A165. After the identification of anti-VEGF-A antibodies, Sanger sequencing and germline mapping were performed to assess the diversity of the antibody candidates. Then, the anti-VEGF-A165 antibodies were cloned into soluble expression vectors and further characterization was performed. Kinetic screening and competition assays were performed to characterize the antibody binding kinetics and affinity to the target.

[0123] Once we narrowed down the antibody candidate pool to eight candidates, in vitroDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 angiogenesis assays were performed to evaluate the ability of the antibody candidates to block endothelial cell proliferation and tube formation. A common hallmark of angiogenesis is the proliferation, migration, and formation of vessel networks by endothelial cells. Thus, primary human umbilical vascular endothelial cells (HUVECs) are commonly used in angiogenesis studies as a method to model vessel normalization and screen potential anti-angiogenic agents. To analyze the ability of the antibody candidates to inhibit angiogenesis, a HUVEC proliferation assay and tube formation assay was performed (Stryker et al., 2019). The results from the HUVEC proliferation assay and tube formation assay allowed us to identify a high affinity anti-VEGF-A165 antibody with biological function comparable to Bevacizumab.

[0124] Presently, we are setting up a HUVEC migration assay to continue to validate the anti-angiogenic activity of the lead antibody candidate (E5-F3) in vitro. Secondly, we are performing the steps to design a CAR-T cell with the lead antibody candidate as the payload.

[0125] Materials and Methods

[0126] Discovery of anti-VEGF-A antibodies

[0127] Phage display library collection

[0128] Phage single chain variable fragment (scFv) libraries were generated with isolated human B cells from healthy donors. The phage display library was incubated in immunotubes coated with hVEGF-A165 (Biolegend, Cat No. 583706) overnight. All unbound phage were washed out extensively with phosphate-buffered saline (PBS) with Tween (PBS and 0.05% Tween-20X). Then, the phage bound to the immunotubes was eluted and neutralized using 1 M Tris – HCl. Additional phage were produced via infection in TG1 E. coli cells. The recovered phage was further enriched with additional rounds of panning against hVEGF-A165 at lower concentrations (Alfaleh et al., 2020).

[0129] Small-scale phage rescue and screening of phagemid antibodies

[0130] Screening of the phage display library was performed after the third round of panning based on a protocol written by the lab. Single bacterial colonies from the third round of panning were picked and cultured in a 96 well-plate overnight in 150 µl of 2YTGA (1% Glucose and 100 µg / ml Ampicillin). After overnight incubation, 50 µl of culture from the 96 well plates were added to 96 deep well plates which contained 0.7 mL of 2YTGA. The cultures in the deep well plate were grown for 3 hours shaking at 37°C. Then, 50 µl of YT media containing 5 x 109 of helper phage was added to each well. The plate was left to stand for 30Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 minutes at 37°C to facilitate the infection of the helper phage into TG1 bacteria. The plates were spun at 3,000 rpm for 10 minutes and then resuspended with 0.8 mL of 2YTAK (Ampicillin 100 ug / ml, Kanamycin 50 ug / ml). The plates were grown overnight with shaking at 37°C. The next day, the plates were spun at 3,000 rpm for 20 minutes and the supernatants were collected to be used for a phage enzyme-linked immunoassay (ELISA) protocol.

[0131] Recombinant human VEGF-A165 (Biolegend, Cat. No. 798206) coating protein was prepared at 2 µg / mL in PBS. Then, 100 µl of the coating protein in PBS was added to Maxisorb U-bottom plates (Nunc #449824). The plate was washed with PBS and 2% Bovine Serum Albumin (BSA) + PBS was used to block the plate. After a one-hour incubation and subsequent wash in PBS, 50 µl of phagemid supernatant and 50 µl 4% milk / 2x PBS-T was added to each well and incubated for an hour. The plates were washed six times with PBS-T. Next, anti-M13-HRP antibody (Sigma, Product #27-9421-01) was diluted in 2% milk / 2x PBS- T at 1:5000 and 100 µl of this solution was added to each well. The plates were washed six times with PBS-T. Finally, 100 µl of TMB substrate was added to each well. When the plates developed, 100 µl of 1M H2SO4 stop solution was added and the plate was read on the Bio- Rad microplate reader at 450 nm. Positive wells were selected for Sanger sequencing and germline mapping was performed (Quintara Biosciences).

[0132] In-Fusion cloning of phage scFv to scFv-Fcs

[0133] First, pFarber plasmids were isolated from TG1 cells. Then, PCR with Vent polymerase was performed on the scFvs using universal primers Sfil-Fwd, Kconst-Rev, and Lconst-Rev (Genewiz). The Vent Polymerase protocol was adapted from New England Biolabs. Cloning enhancer (2 µl) was added to 5 µl of the PCR product and incubated for 20 minutes at 37°C. The pcDNA3.4-stuffer-Fc vector was linearized with Sfil and Not1 restriction enzyme sites. The next day, In-Fusion cloning was performed using our linearized vector, cloning enhancer-treated PCR product, ddH20 (added only to bring the reaction to 8µl volume), and 2µl of 5x In-fusion enzyme mix. Transformation of competent E. coli was performed in a 96-well plate and the plate was incubated shaking overnight so the colonies could grow. The next day, a miniprep was performed using the QIAprep Spin Miniprep Kit (QIAGEN) to extract DNA from our bacteria.

[0134] Characterization of anti-VEGF-A Antibodies

[0135] Protein A Purification and SDS PAGEDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0136] Expi293F cells were transfected with antibody candidate minipreps following the Thermofisher Gibco protocol. A Human IgG ELISA Quantification was performed (Bethyl Laboratories, Cat. No. E80-104) to confirm the successful transfection of plasmid DNA. To perform protein purification, Expi293F cells were centrifuged at 4,000g for 10 minutes. Then, resin was washed with binding buffer three times. The Expi293F cell supernatant was transferred to a new 50 mL tube and 300 µl of resin was added. The mixture was incubated overnight at 4°C. The next day, the resin was transferred to a 20 ml column (BioRad Poly-Prep Chromatography Columns #7311550). The tube was washed was supernatant that flowed through the column once. Then, the columns were washed with 30 ml of PBS three times. Next, 2 ml of IgG Elution Buffer (Thermo #21009) was added to the columns for a five-minute incubation. For protein A purification, 1 M Tris-HCL pH 8 was added as a neutralization buffer. Once a final elution volume of 10 mL was reached, the concentration was recorded on the Nanodrop.

[0137] An SDS page was run to confirm the size of the antibody candidates. Invitrogen NuPage 4-12% Bis-Tris Plus SDS-Page gels were used (Lot 20010970). The loading dye (Invitrogen) was mixed with 2 µg of each protein at a 1:4 dilution. Then, 20 µl of the mixture was loaded per well. The gel was imaged, and the size of the proteins was confirmed.

[0138] Biolayer Interferometry kinetic and competition binding assays

[0139] Binding kinetic assays were performed on the ForteBio Octet RED96 platform. Biotinylated recombinant human VEGF-A165 (Biolegend, Cat. No.522503) was immobilized onto a biotinylated streptavidin sensor. Then, the anti-VEGF-A antibody candidates were associated with the sensor. The antibody candidates were presented in different concentrations ranging from 25 nM to 6.25 nM using a 2x dilution. The sensor was placed in PBS-T to measure the dissociation, or Koff, of the antibody candidates. The sensor was regenerated in 0.1M glycine- HCl so that further analysis could be performed for the antibody candidates. The Kon and Koff generated a KD value for the antibodies that allowed us to evaluate the kinetic profiles of the constructs (Petersen, 2017).

[0140] Epitope binning of our anti-VEGF-A antibody constructs was measured by immobilizing biotinylated VEGF-A on a biotinylated sensor and saturating with antibody candidates or Bevacizumab (AvastinTM, Leinco Technologies Prod. No.: LT400). Our candidates and Bevacizumab were used to saturate VEGF-A loaded on the surface of aDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 biotinylated streptavidin sensor and competition for binding was tested (Petersen, 2017). In both the kinetic and competitive binding assays, a 96-well black plate was used. Additionally, the kinetics and competition were recorded on the OctetRed96. The percent of binding for our competition binding assay was calculated as: % = 100 * (signal of saturating and competing scFv-Fc) / (signal of PBS-T).

[0141] In vitro Functional Assays with Antibody Candidates

[0142] Cell Culture

[0143] Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza and cultured in EBM™-2 Basal Medium (Lonza, Cat. No. CC-3156) supplemented with EGM™- 2 SingleQuots Supplements (Lonza, Cat. No. CC-4176). The EBM-2 media was replaced every 2-3 days and the HUVECs were passaged when they reached 80% confluence. All assays were performed with HUVECs that were less than 15 passages.

[0144] Quantification of secreted VEGF-A165

[0145] TC-32, Skrc-59, CADO-ES1, and A549 cells were donated by members in the lab to measure VEGF-A secretion. The cells were seeded in a 6-well plate at 1 x 105cells per well. Once the cells were ~80% confluent, the supernatants were collected daily over four days. Approximately 100 µl of supernatant was collected from each well and transferred to an eppendorf tube. The supernatant was centrifuged at 13,000 rpm for 5 minutes to remove any debris. Then, the supernatant was transferred to a plate to be stored for future use. After four days of collection, the VEGF-A secretion was measured using the Human VEGF ELISA MAX Deluxe Set (Biolegend, Cat. No. 446504). The 96-well plates were coated with a mouse monoclonal anti- human VEGF antibody. The supernatant samples were incubated for an hour. RPMI and DMEM were used as controls. A detection antibody was added to each well and incubated for an hour. During all incubations after the blocking step, the plate was incubated with shaking. Finally, the plate was washed 5 times with PBS-T and 100 µl of TMB was added to each well. Once the plate developed and the wells turned blue, TMB stop solution was added immediately. The plate was read at 450 nm.

[0146] ELISA VEGF binding assay

[0147] ELISA plates were coated with soluble proteins VEGF-A121, VEGF-A165, VEGF- C, or VEGF-D (Biolegend) in PBS overnight at 4°C. The plates were blocked with 250 µl 2% BSA in PBS. Then, Bevacizumab and antibody candidate E5-F3 were added at an eight-foldDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 dilution from 8 µg / ml using 2% BSA and PBS-T. After an hour, the plates were washed 5 times with PBS-T and incubated with Goat anti-Human IgG-Fc Fragment HRP Conjugated (Bethyl, Catalog No. NC9873055). After an hour of incubation, the plate was washed 5 times with PBS- T, and 100 µl of TMB was added to each well. Once the plate developed and the wells turned blue, TMB stop solution was added immediately. The plate was read at 450 nm.

[0148] HUVEC proliferation assay

[0149] HUVECs were grown in a T-75 flask until they reached approximately 65-70% confluency. Around 16-20 hours before the assay, the media was aspirated from each well and 15 mL of M199 starvation media (5% FBS, Gibco, Cat. No.11150059) was added to each T- 75 flask. After overnight starvation, the HUVECs were passaged and seeded in a 96-well imaging plate (Thermofisher) at a density of 5,000 cells per well. The media used for the assay was M199 starvation media. For antibody and Bevacizumab inhibition experiments, 60 nM of Bevacizumab or antibodies were incubated with 50 ng / ml of recombinant human VEGF-A165 (Biolegend, Cat No.583706) for 2 hours at 37°C. The HUVECs were incubated with the VEGF + Bevacizumab or VEGF + antibody mixture for 72 hours. To measure proliferation, the HUVECs were rinsed with PBS and stained with Hoechst 33342 (Invitrogen, Cat. No. H3570) dye for 30 minutes. After the staining incubation, the plate was read on the Celigo Image Cytometer. Percent growth was calculated as % = (A-B) / B x 100, where A is the number of cells in the VEGF-only condition and B is the number of cells in the M199 media-only condition (Hein & Graver, 2013; Jin et al., 2011).

[0150] HUVEC tube formation assay

[0151] The day before the assay, Corning Matrigel Matrix (Corning, Cat. No.354234) was thawed on ice at 4°C. To prepare the plates, 0.3 mL of chilled Corning Matrigel Matrix was added per well in a 24-well plate. The plates were incubated at 37°C for an hour. The HUVECs were passaged and then seeded onto the Matrigel-coated plate at 4 x 105 cells / mL. The assay media used was M199 media except for one group that tested tube formation with EBM-2 media (with EGM-2 supplements). In each well, hVEGF-A (30 ng / mL) was added to either Bevacizumab (15 µg / mL), E5-F3 (10.5 ug / mL), an irrelevant control antibody (11.2 mg / mL), or M199 media. The plate was incubated for 16 to 18 hours at 37 °C (Carneiro, 2009). The wells were imaged at 5x to 10x magnification. The images were analyzed using Image J with the Angiogenesis Analyzer plugin to quantify the tube formation (Carpentier et al., 2020).Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0152] Statistical Analysis

[0153] One-way ANOVA was used to determine the statistical significance of the experimental conditions. A p-value of p < 0.05 was determined to be statistically significant. A 0.01< p < 0.05 was denoted by *, a 0.001 < p < 0.01 was denoted by **, a 0.0001 < p < 0.001 was denoted by ***, a p < 0.0001 was denoted by ****. Experiments were analyzed using GraphPad and Image J software.

[0154]

[0155] Results

[0156] The quantification of recombinant hVEGF-A165 secretion was measured from cell lines commonly used in the lab. hVEGF-A165 quantification was performed using the ELISA Max Deluxe Set. The results demonstrated that tumor cells secreted hVEGF-A165 in a time- dependent manner in response to stress-invoking conditions. The response to hVEGF-A165 increased with time since the cells were seeded. Additionally, the results demonstrated that the human renal cell carcinoma Skrc-59++ cells produced the highest levels of hVEGF-A165 (FIG. 2).

[0157] Discovery and Characterization of anti-VEGF-A Antibodies

[0158] The aim of this project was to identify a collection of anti-VEGF-A165 antibodies. Phage biopanning campaigns targeting expressed VEGF-A165 initiated the discovery of new anti-VEGF-A165 antibodies. Approximately 960 colonies were selected and cultured for screening by phage ELISA. Phage ELISA was followed by Sanger sequencing of the positive clones to determine the diversity of our candidates. The sequencing alignment and germline mapping of our positive clones resulted in the discovery of 32 unique clones. The phage scFvs were then cloned into soluble expression vectors using a high-throughput in-fusion cloning method. Of the 32 unique clones, 18 clones were successfully expressed as scFv-Fcs (FIG. 3, panel A). After in-fusion cloning, the 18 scFv-Fcs were transfected in Expi293 cells following Thermofisher’s protocol. Three of the 18 scFv-Fcs failed IgG quantification and were removed from the antibody candidate pool. Antibody candidates with similar germline sequencing were organized into groups and our candidate pool was further narrowed to 12 candidates (FIG. 3, panel B).

[0159] Then, competition binding of our remaining 12 antibody candidates was performed using the supernatant from our Expi293 transfection. The results demonstrated that candidateDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 E5-D4 showed simultaneous binding to VEGF-A with Bevacizumab indicating that these molecules might target different epitopes on the VEGF-A protein. The remainder antibody candidates competed with Bevacizumab for binding to biotinylated VEGF-A165 (FIG. 4). During this stage, antibody candidates were categorized according to factors such as VH germline sequences to allow for further characterization (FIG.5 and FIG.6).

[0160] The remaining eight scFv-Fcs were then purified using protein A. Kinetic screening of the antibody candidates was performed using biolayer interferometry (BLI) to measure the association and dissociation rates of our purified proteins (FIG. 7 and FIG. 8). Antibody candidates E5-F3 and E5-A1 had Koff rates (<1.0E-7 s-1) that were below the detection level of the OctetRed. Additionally, E5-F3 and E5-A1 had the lowest amplitude of binding to biotinylated VEGF-A. Regardless, all eight antibody candidates at this stage were selected for testing in our HUVEC functional assays.

[0161] E5-F3 Inhibits HUVEC Proliferation and Tube Formation

[0162] To test the anti-angiogenic activity of the antibody candidates, a HUVEC proliferation assay was performed (FIG. 9). To compare the ability of the eight antibody candidates to block VEGF-stimulated proliferation, Bevacizumab and an irrelevant anti-SARS- 2 antibody were used as controls. In the HUVEC proliferation assay, a significant inhibition of proliferation was observed when HUVECs were incubated with antibody candidate E5-F3 compared to an irrelevant anti-SARs control. Previous studies have demonstrated that Bevacizumab can significantly inhibit proliferation in endothelial cells (Carneiro, 2009; Ferrara, 2005). As research studies previously found, Bevacizumab was able to significantly decrease VEGF-A-induced proliferation in the HUVEC proliferation assay. Due to the ability of E5-F3 to block VEGF-A-induced proliferation, this antibody candidate was selected as the lead anti-VEGF-A antibody candidate.

[0163] The inhibitory ability of E5-F3 was further evaluated by adding recombinant hVEGF-A165 with a concentration range of Bevacizumab, E5-F3, or an irrelevant anti-SARS- 2 antibody for the HUVEC proliferation assay. The results demonstrated that Bevacizumab and E5-F3 resulted in a dose-dependent suppression of HUVEC proliferation after stimulation with hVEGF-A165. Higher concentrations of Bevacizumab and E5-F3 resulted in greater suppression of HUVEC proliferation. As expected, an irrelevant anti-SARS-2 antibody was not able to inhibit VEGF-A- induced HUVEC proliferation (FIG.10).Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0164] To identify the isoforms or subfamily of VEGF that E5-F3 was able to bind to, soluble proteins were purchased from BioLegend for use in an ELISA. Bevacizumab was also measured for comparison. Research studies have demonstrated that Bevacizumab can bind to all VEGF-A isoforms (Papadopoulos et al., 2012). Similarly, Bevacizumab was able to bind to VEGF-A165 and VEGF-A121 in the ELISA. Additionally, the candidate E5-F3 was able to bind to both VEGF-A121 and VEGF-A165. Bevacizumab and E5-F3 were not able to bind to VEGF-C and VEGF-D (FIG.11). Given that the panning was performed against VEGF-A165, the results demonstrate that E5-F3 could have a broader binding specificity.

[0165] To continue to evaluate the anti-angiogenic ability of E5-F3, a tube formation assay was conducted using HUVECs seeded on a Matrigel matrix to mimic the formation of capillary-like structures in vitro (Murray & Martin, 2009). Treatment with E5-F3 demonstrated inhibition of tube formation in VEGF-A-treated HUVECs (FIG. 12). Tube formation was quantified using an Angiogenesis Analyzer plug-in within Image J software. More specifically, the number of junctions and nodes was calculated for each of the experimental groups to quantify the extent of angiogenesis. Junctions were defined as a group of nodes that formed a bifurcation (the location where two points diverge). Nodes were defined as pixels that had a minimum of three adjacent pixels (Li et al., 2023). E5-F3 was able to successfully inhibit the number of junctions and nodes formed compared to the VEGF-A only group. Following previous research studies, Bevacizumab was able to inhibit angiogenesis in comparison to the VEGF-A only control group (Wang et al., 2004). An analysis of the number of nodes found that E5-F3 inhibited the number of nodes formed more significantly than Bevacizumab (FIG. 12).

[0166] Discussion

[0167] One of the goals of this project was to identify and discover an anti-VEGF-A antibody with biological function for tumor vasculature normalization. This field is worthy of attention as clinical study results have demonstrated that a blockage of VEGF-A signaling, through anti- VEGF-A or anti-VEGFR-2 antibodies, can restructure the abnormal tumor vasculature into a normalized vasculature (Yang et al., 2021). A normalized vasculature can lead to improved drug delivery and thus enhanced delivery of immunotherapy (Ager et al., 2016). In this project, we demonstrated that the lead antibody candidate (E5-F3) was effective in inhibiting angiogenesis in the presence of recombinant hVEGF-A in vitro.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025

[0168] A phage panning campaign was used for the enrichment of anti-VEGF phage. Isolation of the anti-VEGF-A antibodies via phage screening allowed for the discovery of 32 unique anti- VEGFA165 antibody clones. Germline mapping and Sanger sequencing allowed us to sort the candidates into antibody families while also providing insight into the diversity of the antibody candidates. Kinetic characterization and BLI competition assays are important assays for antibody discovery as they provide insight into the affinity of antibodies toward their targets and information about the potential epitopes antibodies may bind to. It was found that candidate E5-D4 displayed simultaneous binding to VEGF-A and Bevacizumab. Thus, we could infer that E5-D4 likely targets a different epitope than Bevacizumab does (Sivasubramanian et al., 2017).

[0169] HUVEC tube formation and proliferation assays were performed to allow us to determine if our antibodies displayed desired biological functions. Given that we had discovered anti-VEGF-A165 antibodies, we were expecting to see that our antibodies exhibited anti-angiogenic activity in vitro in response to human rVEGFA-165. Firstly, our HUVEC proliferation assay revealed that endothelial cell proliferation in the presence of hVEGF-A was significantly inhibited with the addition of antibody candidate E5-F3. The other antibody candidates did not block HUVEC proliferation when compared to an irrelevant anti-SARS-2 antibody control. Thus, our decision to select E5-F3 as our lead antibody candidate was based on the superior ability of E5-F3 to inhibit VEGF-induced proliferation of HUVECs in comparison with our other candidates. To further characterize the performance of our lead candidate E5-F3, the HUVEC proliferation assay was repeated with an antibody dilution of E5-F3, Bevacizumab, and an irrelevant anti-SARS-2 antibody. Given that E5-F3 and Bevacizumab both inhibited HUVEC proliferation in a dose- dependent manner, this provided further support for the selection of E5-F3 as our lead antibody candidate. Finally, in a HUVEC tube formation assay E5-F3 inhibited capillary-like tube formation comparably to Bevacizumab. Our findings follow previously published literature demonstrating the ability of Bevacizumab to block angiogenesis in vitro and in vivo (Cheng et al., 2023; Liu et al., 2014).

[0170]

[0171] Alternatives

[0172] Angiogenesis can be measured in vitro primarily through proliferation, tube formation, and migration assays (Stryker et al., 2019; Wang et al., 2004). VEGF-A signalingDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 to VEGFR-2 leads to endothelial cell migration, proliferation, and tube formation (McMahon, 2000). Thus, angiogenesis assays are crucial for evaluating the biological activity of anti- angiogenic agents. The ability of E5-F3 to inhibit endothelial cell proliferation and capillary- like tube formation was successfully tested. Further, the inclusion of a HUVEC scratch assay can validate that E5-F3 can successfully block endothelial cell migration. More specifically, a HUVEC scratch assay entails the “wounding” of a cell monolayer using a pipette tip. Under the presence of VEGF-A, the HUVECs will be stimulated to migrate to the scratch area. However, without wishing to be bound by theory, if E5-F3 or Bevacizumab was added to the wells, HUVEC migration and wound healing will be inhibited (Liu et al., 2014; Nowak- Sliwinska et al., 2018).

[0173] Another assay to test HUVEC migration is the Transwell chamber assay. In this experiment, HUVECs are seeded in the upper chamber of a Transwell insert and the lower chamber contains media supplemented with chemoattractants, such as VEGF-A. To access the anti-angiogenic activity of E5-F3, our antibody candidate or Bevacizumab would be added to the lower chamber. Without wishing to be bound by theory, Bevacizumab and E5-F3 may effectively block HUVEC transmigration (Prager et al., 2010).

[0174] The observed inhibition of HUVEC tube formation and proliferation indicates that E5-F3 is interfering with the angiogenesis process. To validate that E5-F3 is blocking VEGF- A binding to VEGFR-2, we can use a competition binding assay including VEGFR-2 to validate E5-F3 and Bevacizumab (control) can block VEGF-A binding to VEGFR-2.

[0175] Future Directions

[0176] The abnormal tumor microenvironment remains an obstacle to the treatment of solid cancer tumors. A solution to improve CAR-T cell therapy in solid tumors is through the targeting of the tumor vasculature. Our main future direction will be to engineer a CAR-T cell that can secrete our anti-VEGF-A165 monoclonal antibody. Then, I will characterize the level of anti- VEGF-A secretion of our Immune Restoring CAR-T cells. The prospect of designing CAR-T cells that can secrete an anti-VEGFA165 antibody is a new way to counteract the immunosuppressive TME. Previous research studies have demonstrated that CAR-T cells designed against VEGFR-1 or VEGFR-2 have successfully inhibited the growth of vascularized solid tumors and extended the survival of mice (Ager et al., 2016). Additionally, CAR-T cells that were targeting VEGFR-2 and tumor antigens displayed improved anti-tumorDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 activity than CAR-T cells that targeted VEGFR-2 alone (Chinnasamy et al., 2013). While our CAR-T construct would target VEGF-A, these findings demonstrate how the normalization of the tumor vasculature demonstrates promising prospects in the treatment of solid tumor cancers.

[0177] Although we were able to successfully characterize endothelial cell proliferation and tube formation, performing the scratch assay will allow us to gain a comprehensive understanding of angiogenesis from the three main perspectives: endothelial proliferation, migration, and tube formation (Stryker et al., 2019). At the same time, we will design a CAR- T cell with an anti-VEGF-A165 payload that could normalize the tumor vasculature for the treatment of solid tumor cancers. To generate CAR-T cells, we will need to culture 293T cells for lentiviral transfection. Then, T-cells will be isolated from the blood of healthy human donors and stimulated with CD3 / CD28 beads. Finally, the T cells will be transduced with the CAR-encoding lentivirus and in vitro characterization assays could be performed (Hong et al., 2020).

[0178] In conclusion, this project identified and characterized an anti-VEGF-A antibody (E5-F3) with biological functions for tumor vasculature normalization. Through a systematic approach involving a phage panning campaign, kinetic characterization and competition, and functional assays, we demonstrated the ability of E5-F3 to inhibit endothelial tube formation and proliferation induced by human rVEGF-A165. Our findings identify E5-F3 as a lead candidate in future CAR-T cell experiments.

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[0222] Zhang, C., Liu, J., Zhong, J. F., & Zhang, X. (2017). Engineering CAR-T cells. Biomarker Research, 5(1), 22. ***** EQUIVALENTS

[0223] Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific substances and proceduresDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 described herein. Such equivalents are considered to be within the scope of this invention, and are covered by the following claims.

Claims

Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 What is claimed:

1. An antibody or antigen-binding fragment or variant thereof that specifically binds Vascular Endothelial Growth Factor (VEGF) protein, wherein the antibody or antigen- binding fragment comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the HCVR comprises the complementarity determining regions (CDRs) HCDR1, HCDR2 and HCDR3 and the LCVR comprises CDRs LCDR1, LCDR2 and LCDR3, wherein: a) the amino acid sequence of HCDR1 is GFTFNDYE, the amino acid sequence of HCDR2 is VTARGGTE, the amino acid sequence of HCDR3 is ARSARDSYGGDYFDY, the amino acid sequence of LCDR1 is NIGSKS, the amino acid sequence of LCDR2 is YDS, and the amino acid sequence of LCDR3 is QVWDSSSDHPV; or b) the amino acid sequence of HCDR1 is GFAFRTSA, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ASSSGYFDDAFDI, the amino acid sequence of LCDR1 is NIGSKS, the amino acid sequence of LCDR2 is DDS, and the amino acid sequence of LCDR3 is QVWDGGNDDHVV; or c) the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVSPY, the amino acid sequence of LCDR2 is GAS, and the amino acid sequence of LCDR3 is QQYGSSPLT; or d) the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVGEN, the amino acid sequence of LCDR2 is AAS, and the amino acid sequence of LCDR3 is QQFGRSPLT; or e) the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVSRS, the amino acid sequence of LCDR2 is DAS, and the amino acid sequence of LCDR3 is QQYGSSPLT; or f) the amino acid sequence of HCDR1 is GYSFSNYW, the amino acid sequence of HCDR2 is IYPGDSGT, the amino acid sequence of HCDR3 is ARLGQPWTFDS, the amino acid sequence of LCDR1 is QSVRNN, the amino acid sequence of LCDR2 is SAS, and the amino acid sequence of LCDR3 is QQYGSSPIT; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 g) the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is SSYTITQHRG; or h) the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is ISYTITSIVV; or i) the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is VYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDK, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is SSYTITSIVV; or j) the amino acid sequence of HCDR1 is GYSFTSYW, the amino acid sequence of HCDR2 is IYPGDSDT, the amino acid sequence of HCDR3 is ARQKWETMTASAFDN, the amino acid sequence of LCDR1 is SSDLGGHNF, the amino acid sequence of LCDR2 is DVF, and the amino acid sequence of LCDR3 is SSYTITSIVV; or k) the amino acid sequence of HCDR1 is GGTFKDYS, the amino acid sequence of HCDR2 is IIPMYGST, the amino acid sequence of HCDR3 is AREEEGLYRAFDL, the amino acid sequence of LCDR1 is SIGSKS, the amino acid sequence of LCDR2 is DDT, and the amino acid sequence of LCDR3 is QVWDSSSDHPV; or l) the amino acid sequence of HCDR1 is GFTFSSYS, the amino acid sequence of HCDR2 is ISSSGTYI, the amino acid sequence of HCDR3 is ARDSSSWSRTGGMDV, the amino acid sequence of LCDR1 is SLRRYY, the amino acid sequence of LCDR2 is GKN, and the amino acid sequence of LCDR3 is NSRDSSGNPVV; or m) the amino acid sequence of HCDR1 is GYVFTNYF, the amino acid sequence of HCDR2 is INPNSGGT, the amino acid sequence of HCDR3 is ARERKRGYSYGPLDY, the amino acid sequence of LCDR1 is SLSNFF, the amino acid sequence of LCDR2 is GKN, and the amino acid sequence of LCDR3 is NSRDSNGDLVV; or n) the amino acid sequence of HCDR1 is GFTFDDYA, the amino acid sequence of HCDR2 is LSWNSGSV, the amino acid sequence of HCDR3 is AKGTKDSGASGPYFFDY, the amino acid sequence of LCDR1 is TGAVTSDNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGARV; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 o) the amino acid sequence of HCDR1 is GFSFNNYA, the amino acid sequence of HCDR2 is ISWNSGSI, the amino acid sequence of HCDR3 is AKDPAKFRDGYNSDGFDI, the amino acid sequence of LCDR1 is TGAVTSAFY, the amino acid sequence of LCDR2 is NTD, and the amino acid sequence of LCDR3 is LLSYSGARPV; or p) the amino acid sequence of HCDR1 is GFIFDDYA, the amino acid sequence of HCDR2 is INWISGSI, the amino acid sequence of HCDR3 is ARDNEGLYRGFDY, the amino acid sequence of LCDR1 is GSNIGAPYD, the amino acid sequence of LCDR2 is GDN, and the amino acid sequence of LCDR3 is QSYDSSLSGHVV; or q) the amino acid sequence of HCDR1 is ISGDSVSSGA, the amino acid sequence of HCDR2 is TYYSSKWYN, the amino acid sequence of HCDR3 is GRTLSALGNNWFDP, the amino acid sequence of LCDR1 is TGAVTSDNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGARV; or r) the amino acid sequence of HCDR1 is GFDFSNWG, the amino acid sequence of HCDR2 is IGHDGMSQ, the amino acid sequence of HCDR3 is ARDLNSGYTDR, the amino acid sequence of LCDR1 is TGTVTSTNY, the amino acid sequence of LCDR2 is STT, and the amino acid sequence of LCDR3 is LLSYSGSGV; or s) the amino acid sequence of HCDR1 is GYTFSNYG, the amino acid sequence of HCDR2 is ISGHNGKT, the amino acid sequence of HCDR3 is ARDRMRGLYGMDV, the amino acid sequence of LCDR1 is TGAVTSGNY, the amino acid sequence of LCDR2 is IQP, and the amino acid sequence of LCDR3 is LSYPGARG; or t) the amino acid sequence of HCDR1 is GGTFSSYA, the amino acid sequence of HCDR2 is IIPIFGTA, the amino acid sequence of HCDR3 is ARILGGGMDV, the amino acid sequence of LCDR1 is QSVSSSY, the amino acid sequence of LCDR2 is GAS, and the amino acid sequence of LCDR3 is QQYGSSPWT; or u) the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; or v) the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 w) the amino acid sequence of HCDR1 is GGTFSKDV, the amino acid sequence of HCDR2 is ILPMFGST, the amino acid sequence of HCDR3 is ARVHGALYYGMDV, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is QSYDSSLSGWV; or x) the amino acid sequence of HCDR1 is GYRFITYW, the amino acid sequence of HCDR2 is IYPGDSES, the amino acid sequence of HCDR3 is ARRMGGNDAFDV, the amino acid sequence of LCDR1 is QSISSY, the amino acid sequence of LCDR2 is AAS, and the amino acid sequence of LCDR3 is QQANSLRWT; or y) the amino acid sequence of HCDR1 is GFSFSSYP, the amino acid sequence of HCDR2 is IRSGGSPI, the amino acid sequence of HCDR3 is ARDDLYAFDI, the amino acid sequence of LCDR1 is QDISNY, the amino acid sequence of LCDR2 is DAS, and the amino acid sequence of LCDR3 is QQSYSTPIT; or z) the amino acid sequence of HCDR1 is GFIFNNYW, the amino acid sequence of HCDR2 is IERYGGEI, the amino acid sequence of HCDR3 is ARPSWNSGSYFDY, the amino acid sequence of LCDR1 is GSNIGTYT, the amino acid sequence of LCDR2 is RNS, and the amino acid sequence of LCDR3 is SAWDDSLGGEV; or aa) the amino acid sequence of HCDR1 is GFIFNNYW, the amino acid sequence of HCDR2 is IERYGGEI, the amino acid sequence of HCDR3 is ARPSWNSGSYFDY, the amino acid sequence of LCDR1 is NSNIGSYT, the amino acid sequence of LCDR2 is SNN, and the amino acid sequence of LCDR3 is SAWDDSLGGEV; or bb) the amino acid sequence of HCDR1 is GFTFSDYW, the amino acid sequence of HCDR2 is IERYGAEE, the amino acid sequence of HCDR3 is ARPTFNSGSYFDY, the amino acid sequence of LCDR1 is SSNIGAGYD, the amino acid sequence of LCDR2 is GNS, and the amino acid sequence of LCDR3 is SAWDDSLGGEV; or cc) the amino acid sequence of HCDR1 is GFSLTTRGLA, the amino acid sequence of HCDR2 is VYWDDDK, the amino acid sequence of HCDR3 is AHFGYYYFDS, the amino acid sequence of LCDR1 is SGSIASSY, the amino acid sequence of LCDR2 is EDN, and the amino acid sequence of LCDR3 is QSYGSGNWV; or dd) the amino acid sequence of HCDR1 is GFSLTTRGLA, the amino acid sequence of HCDR2 is VYWDDDK, the amino acid sequence of HCDR3 is AHFGYYYFDS, the amino acid sequence of LCDR1 is SGSIASNF, the amino acid sequence of LCDR2 is EDK, and the amino acid sequence of LCDR3 is QSYDTSTHWV; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 ee) the amino acid sequence of HCDR1 is GFSFRMFG, the amino acid sequence of HCDR2 is ISYDGSND, the amino acid sequence of HCDR3 is ARDLGLLQYYYYGMDV, the amino acid sequence of LCDR1 is SSDVGGYNY, the amino acid sequence of LCDR2 is DVS, and the amino acid sequence of LCDR3 is SSYTSSSTLNWV; or ff) the amino acid sequence of HCDR1 is GFSFTMFG, the amino acid sequence of HCDR2 is ISYDGSND, the amino acid sequence of HCDR3 is ARDLGLLQYYYYGMDV, the amino acid sequence of LCDR1 is SSDVGGYNY, the amino acid sequence of LCDR2 is DVS, and the amino acid sequence of LCDR3 is SSYTSSSTLNWG.

2. The antibody or antigen-binding fragment or variant thereof of claim 1, comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein: a) the amino acid sequence of the LCVR is SYELTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNT ATLTISRVEAGDEADYYCQVWDSSSDHPVFGGGTKLAVL and the amino acid sequence of the HCVR is QVQLVQSGGALVRPGGSLRLSCVASGFTFNDYEMNWVRQAPGTGLEWVSSVTARGGTEYYADSVKGRFT ISRDNSKNTVYLQMNSLRGEDTAVYYCARSARDSYGGDYFDYWGQGTLVTVSS; or b) the amino acid sequence of the LCVR is QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPAQAPVLVVYDDSDRPSGIPERFSGSNSGNT ATLTISRVEAGDEADYYCQVWDGGNDDHVVLGGETKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGSVQPGGSLTLSCAASGFAFRTSAMTWVRQAPGRGLEWMGIVYPGDSDTIYSPSFRGQVT ISADKSISTAYLQWSSLKASDTAMYYCASSSGYFDDAFDIWGQGTMVTVSS; or c) the amino acid sequence of the LCVR is EIVLTQSPATLSLSPGERATLSCRAGQSVSPYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or d) the amino acid sequence of the LCVR is ETTLTQSPVTLSVSPGERVTLSCRASQSVGENLAWYQQKPGQAPRLLMYAASTRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQFGRSPLTFGGGTKVEIKDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISVDKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or e) the amino acid sequence of the LCVR is EIVLTQSPGTLSLSPGERATLSCRASQSVSRSLAWYQQKPGQAPRLLIYDASTRATGIPARFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or f) the amino acid sequence of the LCVR is EIVLTQSPATLSLSPGERATLSCRASQSVRNNLAWYQQKPGQAPRLLIYSASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPITFGQGTRLEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or g) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITQHRGFGGGTKLTVP and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or h) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCISYTITSIVVFGGETKLTVL and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or i) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the amino acid sequence of the HCVR is EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 j) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVT ISVDKSINTAYLQWNSLKASDTALYFCARQKWETMTASAFDNWGQGTTVTVSS; or k) the amino acid sequence of the LCVR is SYELTHPPSVSVAPGKTATMTCGGDSIGSKSLNWYQQKPGQAPVLVVYDDTDRPSGIPERFSGSNSGDT ATLTLSRVEAGDEADYYCQVWDSSSDHPVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVTKPGSSVKVSCRTSGGTFKDYSFSWVRQAPGRGLEWMGGIIPMYGSTDYTKKFQGRVT LTADTSTTTLYMELSSLRSEDTAVYYCAREEEGLYRAFDLWGQGTMVTVSS; or l) the amino acid sequence of the LCVR is SSELTQDPAVSVALGQTVRITCQGDSLRRYYASWYQQKPGQAPVLVFYGKNTRPSGIPDRISGSSSGNT ASLTITGAQAEDEADYYCNSRDSSGNPVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSGTYIYYADSVKGRFT ISRDNAKNSLYLQMNSLGAEDTAVYYCARDSSSWSRTGGMDVWGQGTTVTVSS; or m) the amino acid sequence of the LCVR is SSELTQDPAVSVALGQTVRITCQGDSLSNFFAGWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNT ASLTITGAQAEDEADYYCNSRDSNGDLVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGSELKKPGAPVKVSCKTSGYVFTNYFIHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVT MTRDTSISTAYMELSRLRSDDTAVYYCARERKRGYSYGPLDYWGQGTLVTVSS; or n) the amino acid sequence of the LCVR is QTVVTQDPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVL GGKAALTLSGAQPEDEAEYYCLLSYSGARVFGGETKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVPGRSLRLSCAASGFTFDDYAVHWVRQAPGKGLEWVSGLSWNSGSVGYADSVRGRFTI SRDNAGNSLYLQMNGLRAEDTAFYYCAKGTKDSGASGPYFFDYWGQGTLVTVSS; or o) the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGAVTLTCASSTGAVTSAFYPNWFQQKPGQAPKALIYNTDNKHSWTPARFSGSLL GGKAALTLSGAQPEDEAEYYCLLSYSGARPVFGGGTKLTVLDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 and the amino acid sequence of the HCVR is EVQLVESGGGLVQAGGSLTLSCAASGFSFNNYAMSWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFT ISRDNAKNSLYLQMNTLRAEDTALYYCAKDPAKFRDGYNSDGFDIWGQGTTVTVSS; or p) the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGTGSNIGAPYDVHWYQHLPGTAPKLLIYGDNNRPSGVPDRFSGSKS GTSPSLAISGLRSEDEADYYCQSYDSSLSGHVVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGLVQPGRSLRLSCAASGFIFDDYAMHWVRQAPGKGLEWVSGINWISGSIGYADSVKGRFT VSRDNAKNSLYLQMNSLRSEDTAVYYCARDNEGLYRGFDYWGQGTLVTVSS; or q) the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVL GGKAALTLSGAQPEDEAEYYCLLSYSGARVFGGGTKLTVL and the amino acid sequence of the HCVR is MAQVQLQQSGPGLVKPSQTLSLTCGISGDSVSSGAWNWIRSPSRGLQWLGRTYYSSKWYNDYAESVKSR ISINADTSKNQFSLHLNSVTPEDTAVYYCGRTLSALGNNWFDPWGQGTLVTVSS; or r) the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGTVTLTCTSSTGTVTSTNYPNWFQQKPGQAPRPLIYSTTKRHSWTPARFSGSLL GGKAALTLSGAQPEDEAEYFCLLSYSGSGVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFDFSNWGMHWIRQAPGKGLEGVAVIGHDGMSQRYADSVKGRFT VSRDNSKNTQYLEMNSLRVEDTALYYCARDLNSGYTDRWGQGTLVTVSS; or s) the amino acid sequence of the LCVR is QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSGNYPNWFQQKPGQAPRPLIYIQPKDTPGPLPGSQAPPW GQSCPDTVRCAPEDEADYYCLLSYPGARGLRTGTNLTVL and the amino acid sequence of the HCVR is QVQLVQSGAEVNKPGASVKVSCKASGYTFSNYGLTWMRQAPGQGLEWMGWISGHNGKTLSAQKFQDRLL MTTDTSTTTAWLELRSLRSDDTAVYYCARDRMRGLYGMDVWGQGTMVTVSS; or t) the amino acid sequence of the LCVR is EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSG TDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK and the amino acid sequence of the HCVR is QVQLQQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVT ITADESTSTAYMELSSLRSEDTAVYYCARILGGGMDVWGQGTLVTVSS; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 u) the amino acid sequence of the LCVR is QSVLTHPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the amino acid sequence of the HCVR is VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or v) the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the amino acid sequence of the HCVR is VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or w) the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGETKLTVL and the amino acid sequence of the HCVR is VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or x) the amino acid sequence of the LCVR is DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT DFTLTISSLQPEDFATYYCQQANSLRWTFGQGTKVEIK and the amino acid sequence of the HCVR is QVQLVQSGAEVKKPGESLRISCQGSGYRFITYWIGWVRQTPGKGLEWMGAIYPGDSESTYSPPFQGQVT MSVDKSINTAYLQWSSLKASDTATYYCARRMGGNDAFDVWGQGTLVTVSS; or y) the amino acid sequence of the LCVR is DIVMTQTPPSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFTGSGSGT DFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIK and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCVASGFSFSSYPMNWVRQAPGKGLEWISHIRSGGSPISYADSVKGRFT ISRDNAKNSLYLQMNSLRAEDTGVYYCARDDLYAFDIWGQGTMVTVSS; or z) the amino acid sequence of the LCVR is LPVLTQPPSASGTPGQRVTISCSGSGSNIGTYTVNWYQQLPGTAPKLLIYRNSQRPSGVPARFSASKSG TSASLAISGLRSEDEADYFCSAWDDSLGGEVFGTGTKVNVLDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 and the amino acid sequence of the HCVR is QVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFT ISRDNAKNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or aa) the amino acid sequence of the LCVR is LPVLTQPPSASGTPGQRVTISCSGSNSNIGSYTVNWYQQFPGTAPKLLIYSNNQRPSGVPDRFSGSKSG TSASLAISGLRFEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFT ISRDNAKNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or bb) the amino acid sequence of the LCVR is QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYFSAWDDSLGGEVFGTGTKVNVL and the amino acid sequence of the HCVR is EVQLVQSGGGLVQPGGSLRLSCEASGFTFSDYWMGWVRQAPGKGLEWVANIERYGAEEKYVDSVRGRFT ISRDNAKNLLYLHMDSLRAEDTAMYYCARPTFNSGSYFDYWGQGTLVTVSS; or cc) the amino acid sequence of the LCVR is NFMLTQPHSVSESPGETVTISCTRSSGSIASSYVQWYQQRPGSSPTTVVYEDNQRPSGVPDRFSGSIDS SSNSASLTISGLKTEDEADYYCQSYGSGNWVFGGGTKLTVL and the amino acid sequence of the HCVR is QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRL SISKDSSKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or dd) the amino acid sequence of the LCVR is NFMLTHPHSVSGSPGETVTISCTRNSGSIASNFVQWYQQRPGGSPKNVIHEDKRRPSGVPDRFSGSIDT SSNSAFLTISGLKTDDEADYYCQSYDTSTHWVFGGGTRLTVL and the amino acid sequence of the HCVR is QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRL SISKDSSKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or ee) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNRFSGSKS GNTASLTISGLQAEDEADYYCSSYTSSSTLNWVFGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFSFRMFGLHWVRQAPGKGLEWVAFISYDGSNDYYADSVKGRFT ISRDNSKNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 ff) the amino acid sequence of the LCVR is QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNLFSGSKS GNTASLTISGLQAEDEADYYCSSYTSSSTLNWGCGGGTKLTVL and the amino acid sequence of the HCVR is QVQLVQSGGGVVQPGRSLRLSCAASGFSFTMFGLHWVRQAPGKGLEWVAFISYDGSNDYYAHSVKGRFT ISRDNSKNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS.

3. The antibody or antigen-binding fragment thereof of claim 1, comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein: a) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SYELTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNT ATLTISRVEAGDEADYYCQVWDSSSDHPVFGGGTKLAVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGALVRPGGSLRLSCVASGFTFNDYEMNWVRQAPGTGLEWVSSVTARGGTEYYADSVKGRFT ISRDNSKNTVYLQMNSLRGEDTAVYYCARSARDSYGGDYFDYWGQGTLVTVSS; or b) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPAQAPVLVVYDDSDRPSGIPERFSGSNSGNT ATLTISRVEAGDEADYYCQVWDGGNDDHVVLGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGSVQPGGSLTLSCAASGFAFRTSAMTWVRQAPGRGLEWMGIVYPGDSDTIYSPSFRGQVT ISADKSISTAYLQWSSLKASDTAMYYCASSSGYFDDAFDIWGQGTMVTVSS; or c) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPATLSLSPGERATLSCRAGQSVSPYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 d) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence ETTLTQSPVTLSVSPGERVTLSCRASQSVGENLAWYQQKPGQAPRLLMYAASTRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQFGRSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISVDKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or e) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPGTLSLSPGERATLSCRASQSVSRSLAWYQQKPGQAPRLLIYDASTRATGIPARFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or f) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPATLSLSPGERATLSCRASQSVRNNLAWYQQKPGQAPRLLIYSASSRATGIPDRFSGSGSGT DFTLTISRLEPEDFAVYYCQQYGSSPITFGQGTRLEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPGDSGTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDSAMYYCARLGQPWTFDSWGQGTLVTVSS; or g) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITQHRGFGGGTKLTVP and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or h) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequenceDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCISYTITSIVVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or i) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIVYPGDSDTRYSPSFQGQVT ISADRSTNTAYLQWSSLKASDTAMYFCARQKWETMTASAFDKWGQGTTVTVSS; or j) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDLGGHNFVSWYQQHPGKAPKLMIYDVFNRPSGVSSRFSGSKS GNTASLTISGLQAEDEADYFCSSYTITSIVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVT ISVDKSINTAYLQWNSLKASDTALYFCARQKWETMTASAFDNWGQGTTVTVSS; or k) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SYELTHPPSVSVAPGKTATMTCGGDSIGSKSLNWYQQKPGQAPVLVVYDDTDRPSGIPERFSGSNSGDT ATLTLSRVEAGDEADYYCQVWDSSSDHPVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVTKPGSSVKVSCRTSGGTFKDYSFSWVRQAPGRGLEWMGGIIPMYGSTDYTKKFQGRVT LTADTSTTTLYMELSSLRSEDTAVYYCAREEEGLYRAFDLWGQGTMVTVSS; or l) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SSELTQDPAVSVALGQTVRITCQGDSLRRYYASWYQQKPGQAPVLVFYGKNTRPSGIPDRISGSSSGNT ASLTITGAQAEDEADYYCNSRDSSGNPVVFGGGTKLTVLDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSGTYIYYADSVKGRFT ISRDNAKNSLYLQMNSLGAEDTAVYYCARDSSSWSRTGGMDVWGQGTTVTVSS; or m) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence SSELTQDPAVSVALGQTVRITCQGDSLSNFFAGWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNT ASLTITGAQAEDEADYYCNSRDSNGDLVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGSELKKPGAPVKVSCKTSGYVFTNYFIHWVRQAPGQGLEWMGRINPNSGGTNYAQKFQGRVT MTRDTSISTAYMELSRLRSDDTAVYYCARERKRGYSYGPLDYWGQGTLVTVSS; or n) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQDPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVL GGKAALTLSGAQPEDEAEYYCLLSYSGARVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVPGRSLRLSCAASGFTFDDYAVHWVRQAPGKGLEWVSGLSWNSGSVGYADSVRGRFTI SRDNAGNSLYLQMNGLRAEDTAFYYCAKGTKDSGASGPYFFDYWGQGTLVTVSS; or o) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGAVTLTCASSTGAVTSAFYPNWFQQKPGQAPKALIYNTDNKHSWTPARFSGSLL GGKAALTLSGAQPEDEAEYYCLLSYSGARPVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVESGGGLVQAGGSLTLSCAASGFSFNNYAMSWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFT ISRDNAKNSLYLQMNTLRAEDTALYYCAKDPAKFRDGYNSDGFDIWGQGTTVTVSS; or p) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGTGSNIGAPYDVHWYQHLPGTAPKLLIYGDNNRPSGVPDRFSGSKS GTSPSLAISGLRSEDEADYYCQSYDSSLSGHVVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequenceDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 QVQLVQSGGGLVQPGRSLRLSCAASGFIFDDYAMHWVRQAPGKGLEWVSGINWISGSIGYADSVKGRFT VSRDNAKNSLYLQMNSLRSEDTAVYYCARDNEGLYRGFDYWGQGTLVTVSS; or q) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSDNYPNWFQQKPGQAPRPLIYSTTKRHAWTPARFSGSVL GGKAALTLSGAQPEDEAEYYCLLSYSGARVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence MAQVQLQQSGPGLVKPSQTLSLTCGISGDSVSSGAWNWIRSPSRGLQWLGRTYYSSKWYNDYAESVKSR ISINADTSKNQFSLHLNSVTPEDTAVYYCGRTLSALGNNWFDPWGQGTLVTVSS; or r) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCTSSTGTVTSTNYPNWFQQKPGQAPRPLIYSTTKRHSWTPARFSGSLL GGKAALTLSGAQPEDEAEYFCLLSYSGSGVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFDFSNWGMHWIRQAPGKGLEGVAVIGHDGMSQRYADSVKGRFT VSRDNSKNTQYLEMNSLRVEDTALYYCARDLNSGYTDRWGQGTLVTVSS; or s) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QTVVTQEPSLTVSPGGTVTLTCASSTGAVTSGNYPNWFQQKPGQAPRPLIYIQPKDTPGPLPGSQAPPW GQSCPDTVRCAPEDEADYYCLLSYPGARGLRTGTNLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVNKPGASVKVSCKASGYTFSNYGLTWMRQAPGQGLEWMGWISGHNGKTLSAQKFQDRLL MTTDTSTTTAWLELRSLRSDDTAVYYCARDRMRGLYGMDVWGQGTMVTVSS; or t) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSG TDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLQQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVT ITADESTSTAYMELSSLRSEDTAVYYCARILGGGMDVWGQGTLVTVSS; orDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 u) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTHPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or v) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or w) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYYCQSYDSSLSGWVFGGETKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence VQLVQSGAEVKKPGSSVKVSCKASGGTFSKDVINWVRQAPGHGLEWMGGILPMFGSTNYAQKFQGRLTL IADESTRTVYLELNSLTSEDTAVYYCARVHGALYYGMDVWGQGTLVTVSS; or x) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT DFTLTISSLQPEDFATYYCQQANSLRWTFGQGTKVEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGAEVKKPGESLRISCQGSGYRFITYWIGWVRQTPGKGLEWMGAIYPGDSESTYSPPFQGQVT MSVDKSINTAYLQWSSLKASDTATYYCARRMGGNDAFDVWGQGTLVTVSS; or y) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequenceDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 DIVMTQTPPSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFTGSGSGT DFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIK and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCVASGFSFSSYPMNWVRQAPGKGLEWISHIRSGGSPISYADSVKGRFT ISRDNAKNSLYLQMNSLRAEDTGVYYCARDDLYAFDIWGQGTMVTVSS; or z) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence LPVLTQPPSASGTPGQRVTISCSGSGSNIGTYTVNWYQQLPGTAPKLLIYRNSQRPSGVPARFSASKSG TSASLAISGLRSEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFT ISRDNAKNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or aa) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence LPVLTQPPSASGTPGQRVTISCSGSNSNIGSYTVNWYQQFPGTAPKLLIYSNNQRPSGVPDRFSGSKSG TSASLAISGLRFEDEADYFCSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCAASGFIFNNYWMGWVRQAPGKGLEWVANIERYGGEIHYADSVQGRFT ISRDNAKNALYLQMNNVRAEDTAVYYCARPSWNSGSYFDYWGQGTLVTVSS; or bb) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKS GTSASLAITGLQAEDEADYFSAWDDSLGGEVFGTGTKVNVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence EVQLVQSGGGLVQPGGSLRLSCEASGFTFSDYWMGWVRQAPGKGLEWVANIERYGAEEKYVDSVRGRFT ISRDNAKNLLYLHMDSLRAEDTAMYYCARPTFNSGSYFDYWGQGTLVTVSS; or cc) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence NFMLTQPHSVSESPGETVTISCTRSSGSIASSYVQWYQQRPGSSPTTVVYEDNQRPSGVPDRFSGSIDS SSNSASLTISGLKTEDEADYYCQSYGSGNWVFGGGTKLTVLDocket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRL SISKDSSKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or dd) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence NFMLTHPHSVSGSPGETVTISCTRNSGSIASNFVQWYQQRPGGSPKNVIHEDKRRPSGVPDRFSGSIDT SSNSAFLTISGLKTDDEADYYCQSYDTSTHWVFGGGTRLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVTLKESGPALVKPTETLTLTCTFSGFSLTTRGLAVGWIRQPPGKALEFLALVYWDDDKRYSPSLKNRL SISKDSSKNQVVLTVTNVDPLDTGTYYCAHFGYYYFDSWGHGTLVTVSS; or ee) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNRFSGSKS GNTASLTISGLQAEDEADYYCSSYTSSSTLNWVFGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFSFRMFGLHWVRQAPGKGLEWVAFISYDGSNDYYADSVKGRFT ISRDNSKNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS; or ff) the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNLFSGSKS GNTASLTISGLQAEDEADYYCSSYTSSSTLNWGCGGGTKLTVL and the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence QVQLVQSGGGVVQPGRSLRLSCAASGFSFTMFGLHWVRQAPGKGLEWVAFISYDGSNDYYAHSVKGRFT ISRDNSKNTLYLQMNRLRPEDTAMYYCARDLGLLQYYYYGMDVWGQGTTVTVSS.

4. The antibody or antigen-binding fragment or variant thereof of any one of claims 1-3, wherein the fragment comprises an F(ab), an Fv, or an scFv.

5. The antibody or antigen-binding fragment or variant thereof of any one of claims 1-3, wherein the fragment comprises a VhH.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 6. A therapeutic antibody that binds to an Vascular Endothelial Growth Factor (VEGF) protein comprising a variable domain and a constant domain, wherein the constant domain is IgG and the variable domain comprises a framework region and a complementary determining means for binding to an Vascular Endothelial Growth Factor (VEGF) protein.

7. The therapeutic antibody of claim 6, wherein the constant region is IgG4.

8. The therapeutic antibody of any one of claims 6-7, wherein the antibody comprises any one of the antibodies according to Table 1 – Table 4.

9. A pharmaceutical composition, comprising: the antibody of any one of claims 1-8, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

10. A method of treating cancer in a subject, the method comprising administering to a subject in need thereof an effective amount of the antibody of any one of claims 1-8, or the pharmaceutical composition of claim 9.

11. The method of claim 10, wherein the cancer comprises Ewing carcinoma, renal cell carcinoma, or lung adenocarcinoma.

12. Use of the antibody of any one of 1-8, or the pharmaceutical composition of claim 9 for treating cancer.

13. The antibody of any one of claims 1-8, or the pharmaceutical composition of claim 9 for use in treating cancer.

14. A method of generating and / or increasing angiogenesis in a cancer in a subject, comprising administering to the subject an effective amount of the antibody of any one of claims 1-8, or the pharmaceutical composition of claim 9.Docket No.: 5031461-000164WO1 Date of Filing: MARCH 10, 2025 15. Use of the antibody of any one of 1-8, or the pharmaceutical composition of claim 9 for increasing angionesis in a cancer.

16. The antibody of any one of claims 1-8, or the pharmaceutical composition of claim 9 for use in increasing angiogenesis in a cancer.

17. A nucleic acid encoding the antibody of claim 1 or claim 2.

18. A vector comprising the nucleic acid of claim 17.

19. A cell comprising the nucleic acid of any one of claim 17 or the vector of claim 18.

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