Anti-programmed death ligand 1 (PD-L1) antibody molecules, encoding polynucleotides and methods of use

Abstract

Provided herein are anti-programmed death ligand (PD-L1) antibodies, antigen binding fragments thereof, multispecific antibodies, and encoding polynucleotides. Also provided are PD-L1 targeted phthalocyanine dye conjugates, compositions and articles of manufacture containing the conjugates, and methods of administering the conjugates to an individual for photoimmunotherapy. In some embodiments, the anti-PD-L1 antibodies, fragments, and conjugates specifically bind to human PD-L1. The anti-PD-L1 antibody comprises a human antibody and a humanized antibody. Methods and uses of the provided anti-PD-L1 antibodies, antigen binding fragments, and conjugates for treating human patients are also provided.

Description

[0001] Related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 311,924, filed on February 18, 2022, entitled “ANTI-PROGRAMMED DEATH-LIGAND 1 (PD-L1) ANTIBODY MOLECULES, ENCODING POLYNUCLEOTIDES, AND METHODS OF USE,” the contents of which are incorporated by reference in their entirety.

[0003] The sequence listing is incorporated by reference

[0004] The contents of the electronic sequence listing (file name: 751702002740SeqList.xml, creation date: February 7, 2023, size: 418,493 bytes) are incorporated herein by reference in their entirety. Technical Field

[0005] The present disclosure relates to programmed death ligand (PD-L1) binding molecules, in particular, to anti-PD-L1 antibodies, including antibody fragments and compositions, combinations, methods, and uses thereof. The present disclosure further relates to conjugates containing such antibodies and compositions, combinations, methods, and uses of such conjugates. The present disclosure further relates to nucleic acid molecules encoding the PD-L1 antibodies and fragments described herein. Background Art

[0006] Programmed death ligand 1 (PD-L1), also known as cluster of differentiation 247 (CD247) or B7-H1, is a protein receptor that serves as an immune checkpoint and downregulates immune responses. Although therapies involving inhibitors of the PD-1:PD-L1 pathway have been used to treat cancer, challenges remain. Improved compositions and methods are urgently needed to address these clinical challenges. Embodiments are provided to address these needs. Summary of the Invention

[0007] Provided herein is an antibody or antigen-binding fragment that specifically binds to a programmed death ligand (PD-L1) protein. In some embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a heavy chain variable (VH) region comprising a heavy chain complementary determining region 1 (CDR-H1), CDR-H2, and CDR-H3, respectively, comprising the CDR-H1, CDR-H2, and CDR-H3 contained within the VH region sequence set forth in SEQ ID NO: 1; and a light chain variable (VL) region comprising a light chain complementary determining region 1 (CDR-L1), CDR-L2, and CDR-L3, respectively, comprising the CDR-L1, CDR-L2, and CDR-L3 contained within the VL region sequence set forth in SEQ ID NO: 17. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 2; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 18. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 3; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 19. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:4; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:19.In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:5; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:20. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 1; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 21. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 2; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 22. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:6; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:238. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:7; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:24.In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:8; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:25. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:9; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:26. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 10; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 27. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 11; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 28. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 12; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 29.In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 13; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 30. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 14; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 31. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 15; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 32. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 12; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 33. In some of any of the embodiments, the PD-L1 binding antibody or antigen-binding fragment comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3, respectively, comprising CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO: 16; and a VL region comprising CDR-L1, CDR-L2, and CDR-L3, respectively, comprising CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO: 34.

[0008] In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 35, a CDR-H2 comprising SEQ ID NO: 36, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 210, a CDR-L2 comprising SEQ ID NO: 211, and a CDR-L3 comprising SEQ ID NO: 212. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 48, a CDR-H2 comprising SEQ ID NO: 49, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 218, a CDR-L2 comprising SEQ ID NO: 211, and a CDR-L3 comprising SEQ ID NO: 212. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 58, a CDR-H2 comprising SEQ ID NO: 59, and a CDR-H3 comprising SEQ ID NO: 60; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 221, a CDR-L2 comprising SEQ ID NO: 222, and a CDR-L3 comprising SEQ ID NO: 223. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 71, a CDR-H2 comprising SEQ ID NO: 72, and a CDR-H3 comprising SEQ ID NO: 60; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 229, a CDR-L2 comprising SEQ ID NO: 222, and a CDR-L3 comprising SEQ ID NO: 223. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 35, a CDR-H2 comprising SEQ ID NO: 36, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 233, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 235. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 48, a CDR-H2 comprising SEQ ID NO: 49, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 241, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 242.In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 82, a CDR-H2 comprising SEQ ID NO: 83, and a CDR-H3 comprising SEQ ID NO: 84; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 247, and a CDR-L3 comprising SEQ ID NO: 248. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 48, a CDR-H2 comprising SEQ ID NO: 95, and a CDR-H3 comprising SEQ ID NO: 84; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 254, and a CDR-L3 comprising SEQ ID NO: 255. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 48, a CDR-H2 comprising SEQ ID NO: 104, and a CDR-H3 comprising SEQ ID NO: 105; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 258, a CDR-L2 comprising SEQ ID NO: 259, and a CDR-L3 comprising SEQ ID NO: 260. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 116, a CDR-H2 comprising SEQ ID NO: 117, and a CDR-H3 comprising SEQ ID NO: 118; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 265, a CDR-L2 comprising SEQ ID NO: 266, and a CDR-L3 comprising SEQ ID NO: 267. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 129, a CDR-H2 comprising SEQ ID NO: 130, and a CDR-H3 comprising SEQ ID NO: 131; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 273, and a CDR-L3 comprising SEQ ID NO: 274. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 142, a CDR-H2 comprising SEQ ID NO: 143, and a CDR-H3 comprising SEQ ID NO: 144; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 278, a CDR-L2 comprising SEQ ID NO: 279, and a CDR-L3 comprising SEQ ID NO: 280.In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 155, a CDR-H2 comprising SEQ ID NO: 156, and a CDR-H3 comprising SEQ ID NO: 157; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 286, a CDR-L2 comprising SEQ ID NO: 287, and a CDR-L3 comprising SEQ ID NO: 288. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 168, a CDR-H2 comprising SEQ ID NO: 169, and a CDR-H3 comprising SEQ ID NO: 170; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 294, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 295. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 35, a CDR-H2 comprising SEQ ID NO: 181, and a CDR-H3 comprising SEQ ID NO: 182; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 299, a CDR-L2 comprising SEQ ID NO: 300, and a CDR-L3 comprising SEQ ID NO: 301. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 168, a CDR-H2 comprising SEQ ID NO: 169, and a CDR-H3 comprising SEQ ID NO: 193; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 306, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 307. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 155, a CDR-H2 comprising SEQ ID NO: 156, and a CDR-H3 comprising SEQ ID NO: 157; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 311, a CDR-L2 comprising SEQ ID NO: 312, and a CDR-L3 comprising SEQ ID NO: 313. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 197, a CDR-H2 comprising SEQ ID NO: 198, and a CDR-H3 comprising SEQ ID NO: 199; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 319, a CDR-L2 comprising SEQ ID NO: 320, and a CDR-L3 comprising SEQ ID NO: 321.

[0009] In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 40, a CDR-H2 comprising SEQ ID NO: 41, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 210, a CDR-L2 comprising SEQ ID NO: 211, and a CDR-L3 comprising SEQ ID NO: 212. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 52, a CDR-H2 comprising SEQ ID NO: 53, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 218, a CDR-L2 comprising SEQ ID NO: 211, and a CDR-L3 comprising SEQ ID NO: 212. In some of any embodiments, the VH region comprises: a CDR-H1 comprising SEQ ID NO: 63, a CDR-H2 comprising SEQ ID NO: 64, and a CDR-H3 comprising SEQ ID NO: 60; and the VL region comprises: a CDR-L1 comprising SEQ ID NO: 221, a CDR-L2 comprising SEQ ID NO: 222, and a CDR-L3 comprising SEQ ID NO: 223. In some of any embodiments, the VH region comprises: a CDR-H1 comprising SEQ ID NO: 75, a CDR-H2 comprising SEQ ID NO: 76, and a CDR-H3 comprising SEQ ID NO: 60; and the VL region comprises: a CDR-L1 comprising SEQ ID NO: 229, a CDR-L2 comprising SEQ ID NO: 222, and a CDR-L3 comprising SEQ ID NO: 223. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 40, a CDR-H2 comprising SEQ ID NO: 41, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 233, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 235. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 52, a CDR-H2 comprising SEQ ID NO: 53, and a CDR-H3 comprising SEQ ID NO: 37; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 241, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 242.In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 87, a CDR-H2 comprising SEQ ID NO: 88, and a CDR-H3 comprising SEQ ID NO: 84; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 247, and a CDR-L3 comprising SEQ ID NO: 248. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 98, a CDR-H2 comprising SEQ ID NO: 99, and a CDR-H3 comprising SEQ ID NO: 84; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 254, and a CDR-L3 comprising SEQ ID NO: 255. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 108, a CDR-H2 comprising SEQ ID NO: 109, and a CDR-H3 comprising SEQ ID NO: 105; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 258, a CDR-L2 comprising SEQ ID NO: 259, and a CDR-L3 comprising SEQ ID NO: 260. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 121, a CDR-H2 comprising SEQ ID NO: 122, and a CDR-H3 comprising SEQ ID NO: 118; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 265, a CDR-L2 comprising SEQ ID NO: 266, and a CDR-L3 comprising SEQ ID NO: 267. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 134, a CDR-H2 comprising SEQ ID NO: 135, and a CDR-H3 comprising SEQ ID NO: 131; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 246, a CDR-L2 comprising SEQ ID NO: 273, and a CDR-L3 comprising SEQ ID NO: 274. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 147, a CDR-H2 comprising SEQ ID NO: 148, and a CDR-H3 comprising SEQ ID NO: 144; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 278, a CDR-L2 comprising SEQ ID NO: 279, and a CDR-L3 comprising SEQ ID NO: 280.In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 160, a CDR-H2 comprising SEQ ID NO: 161, and a CDR-H3 comprising SEQ ID NO: 157; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 286, a CDR-L2 comprising SEQ ID NO: 287, and a CDR-L3 comprising SEQ ID NO: 288. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 173, a CDR-H2 comprising SEQ ID NO: 174, and a CDR-H3 comprising SEQ ID NO: 170; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 294, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 295. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 185, a CDR-H2 comprising SEQ ID NO: 186, and a CDR-H3 comprising SEQ ID NO: 182; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 299, a CDR-L2 comprising SEQ ID NO: 300, and a CDR-L3 comprising SEQ ID NO: 301. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 173, a CDR-H2 comprising SEQ ID NO: 174, and a CDR-H3 comprising SEQ ID NO: 193; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 306, a CDR-L2 comprising SEQ ID NO: 234, and a CDR-L3 comprising SEQ ID NO: 307. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 160, a CDR-H2 comprising SEQ ID NO: 161, and a CDR-H3 comprising SEQ ID NO: 157; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 311, a CDR-L2 comprising SEQ ID NO: 312, and a CDR-L3 comprising SEQ ID NO: 313. In some of any embodiments, the VH region comprises a CDR-H1 comprising SEQ ID NO: 202, a CDR-H2 comprising SEQ ID NO: 203, and a CDR-H3 comprising SEQ ID NO: 199; and the VL region comprises a CDR-L1 comprising SEQ ID NO: 319, a CDR-L2 comprising SEQ ID NO: 320, and a CDR-L3 comprising SEQ ID NO: 321.

[0010] In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 17. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 2; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 18. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 3; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 19. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 4; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 19. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 5; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 20. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 21. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 21; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 22. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 6; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 23. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 7; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 24. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 8; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 25. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 9; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 26. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 10; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 27.In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 11; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 28. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 12; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 29. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 13; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 30. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 14; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 31. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 15; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 32. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 12; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 33. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 16; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 34.

[0011] In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 330; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 335. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 331; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 336. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 332; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 337. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 333; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 338. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 330; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 339. In some of any embodiments, the VH region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 334; and the VL region comprises a sequence having at least 95% sequence identity to SEQ ID NO: 340.

[0012] In some of any embodiments, the VH region comprises SEQ ID NO: 1; and the VL region comprises SEQ ID NO: 17. In some of any embodiments, the VH region comprises SEQ ID NO: 2; and the VL region comprises SEQ ID NO: 18. In some of any embodiments, the VH region comprises SEQ ID NO: 3; and the VL region comprises SEQ ID NO: 19. In some of any embodiments, the VH region comprises SEQ ID NO: 4; and the VL region comprises SEQ ID NO: 19. In some of any embodiments, the VH region comprises SEQ ID NO: 5; and the VL region comprises SEQ ID NO: 20. In some of any embodiments, the VH region comprises SEQ ID NO: 1; and the VL region comprises SEQ ID NO: 21. In some of any embodiments, the VH region comprises SEQ ID NO: 21; and the VL region comprises SEQ ID NO: 22. In some of any embodiments, the VH region comprises SEQ ID NO: 6; and the VL region comprises SEQ ID NO: 23. In some of any embodiments, the VH region comprises SEQ ID NO: 7; and the VL region comprises SEQ ID NO: 24. In some of any embodiments, the VH region comprises SEQ ID NO: 8; and the VL region comprises SEQ ID NO: 25. In some of any embodiments, the VH region comprises SEQ ID NO: 9; and the VL region comprises SEQ ID NO: 26. In some of any embodiments, the VH region comprises SEQ ID NO: 10; and the VL region comprises SEQ ID NO: 27. In some of any embodiments, the VH region comprises SEQ ID NO: 11; and the VL region comprises SEQ ID NO: 28. In some of any embodiments, the VH region comprises SEQ ID NO: 12; and the VL region comprises SEQ ID NO: 29. In some of any embodiments, the VH region comprises SEQ ID NO: 13; and the VL region comprises SEQ ID NO: 30. In some of any embodiments, the VH region comprises SEQ ID NO: 14; and the VL region comprises SEQ ID NO: 31. In some of any embodiments, the VH region comprises SEQ ID NO: 15; and the VL region comprises SEQ ID NO: 32. In some of any embodiments, the VH region comprises SEQ ID NO: 12; and the VL region comprises SEQ ID NO: 33. In some of any embodiments, the VH region comprises SEQ ID NO: 16; and the VL region comprises SEQ ID NO: 34.

[0013] In some of any embodiments, the VH region comprises SEQ ID NO: 330; and the VL region comprises SEQ ID NO: 335. In some of any embodiments, the VH region comprises SEQ ID NO: 331; and the VL region comprises SEQ ID NO: 336. In some of any embodiments, the VH region comprises SEQ ID NO: 332; and the VL region comprises SEQ ID NO: 337. In some of any embodiments, the VH region comprises SEQ ID NO: 333; and the VL region comprises SEQ ID NO: 338. In some of any embodiments, the VH region comprises SEQ ID NO: 330; and the VL region comprises SEQ ID NO: 339. In some of any embodiments, the VH region comprises SEQ ID NO: 334; and the VL region comprises SEQ ID NO: 340.

[0014] In some of any of the embodiments, the PD-L1 protein is a human PD-L1 protein.

[0015] In some of any embodiments, the antibody or antigen-binding fragment is recombinant. In some of any embodiments, the antibody or antigen-binding fragment is monoclonal. In some of any embodiments, the antibody or antigen-binding fragment is human, chimeric, or humanized.

[0016] In some of any of the embodiments, the antibody or antigen-binding fragment comprises an Fc region of a human immunoglobulin and / or a human antibody framework region.

[0017] In some of any embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment. In some of any embodiments, the antibody fragment comprises a single-chain Fv (scFv).

[0018] In some of any of the embodiments, the antibody is a complete or intact antibody.

[0019] In some of any embodiments, the antibody or antigen-binding fragment is a bispecific antibody that further specifically binds to a second antigen. In some of any embodiments, the second antigen is an antigen expressed on a tumor cell or an immune cell. In some of any embodiments, the second antigen is an antigen expressed on an immune cell, the immune cell is a T cell, and the antigen is CD25.

[0020] In some of any embodiments, the antibody or its antigen binding fragment comprises an Fc region that exhibits one or more Fc-mediated effector functions. In some of any embodiments, the antibody or its antigen binding fragment has an Fc region that lacks Fc-mediated effector functions, exhibits significantly reduced Fc-mediated effector functions or does not exhibit significant Fc-mediated effector functions. In some of any embodiments, the antibody or its antigen binding fragment comprises an Fc region that exhibits enhanced Fc-mediated effector functions. In some of any embodiments, the FcR-mediated effector functions are selected from one or more of antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), or complement-dependent cytotoxicity (CDC).

[0021] In some of any of the embodiments, the antibody or antigen-binding fragment comprises an IgG1 Fc region or IgG1 isotype, an IgG2 Fc region or IgG2 isotype, an IgG3 Fc region or IgG3 isotype, or an IgG4 Fc region or IgG4 isotype.

[0022] Also provided is a conjugate comprising any of the provided antibodies or antigen-binding fragments and a heterologous molecule or moiety. In some of any embodiments, the heterologous molecule or moiety is a protein, peptide, nucleic acid, dye, or small molecule. In some of any embodiments, the heterologous molecule or moiety is a cytotoxic agent, toxin, radioisotope, chemotherapeutic agent, lytic peptide, cytokine, or photoactivated dye.

[0023] In some of any embodiments, the photoactivated dye is a phthalocyanine dye. In some of any embodiments, the phthalocyanine dye is a Si-phthalocyanine dye.

[0024] In some of any of the embodiments, the phthalocyanine dye is IR700.

[0025] In some of any of the embodiments, the phthalocyanine dye has the structure of Formula (I):

[0026] or a salt, stereoisomer or tautomer thereof.

[0027] In some of any of the embodiments, the conjugate is activated via irradiation at a wavelength between at or about 600 nm and 850 nm or at about 850 nm to achieve cell killing.

[0028] In some of any of the embodiments, the activated conjugate achieves tumor growth inhibition or killing at a higher level, activity, or potency than the unconjugated antibody.

[0029] In some of any of the embodiments, the antibody or antigen-binding fragment and the moiety are linked directly or indirectly via a linker.

[0030] In some of any of the embodiments, the antibody or antigen-binding fragment is covalently linked to a heterologous molecule or moiety.

[0031] In some of any embodiments, when contacted with cells expressing the PD-L1 protein, the conjugate exhibits increased internalization compared to an unconjugated antibody or antigen-binding fragment, or a conjugate comprising a reference antibody. In some of any embodiments, when contacted with cells expressing the PD-L1 protein, the conjugate exhibits decreased internalization compared to an unconjugated antibody or antigen-binding fragment, or a conjugate comprising a reference antibody. In some of any embodiments, the reference antibody is avelumab.

[0032] In some of any embodiments, the conjugate does not exhibit significantly reduced binding affinity for the PD-L1 protein compared to the unconjugated antibody, or exhibits at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% of the binding affinity of the unconjugated antibody for the PD-L1 protein. In some of any embodiments, the conjugate exhibits similar binding to the PD-L1 protein compared to the unconjugated antibody.

[0033] Also provided is a polynucleotide encoding any of the provided antibodies or antigen-binding fragments.

[0034] Also provided are vectors comprising any of the provided polynucleotides.In some of any of the embodiments, the vector is an expression vector.

[0035] Also provided are engineered cells comprising any of the provided vectors.

[0036] Also provided are engineered cells that express any of the provided antibodies or antigen-binding fragments.

[0037] Also provided are compositions comprising any of the provided antibodies or antigen-binding fragments or any of the provided conjugates. In some of any of the embodiments, the composition also comprises a pharmaceutically acceptable excipient.

[0038] Also provided is a method for treating a disease or condition. Also provided is any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions for use in any of such methods. Also provided is a use of any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions for use in the manufacture of a medicament for treating a disease or condition. In some of any of the embodiments, the method involves administering any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions to an individual suffering from a disease or condition.

[0039] Also provided is a method of treatment comprising administering any of the provided compositions to an individual having a disease or condition. In some of any of the embodiments, the disease or condition is a tumor or cancer.

[0040] Also provided is a method of treating a tumor or lesion in a subject. In some of any of the embodiments, the method involves administering to the subject any of the provided compositions or any of the provided compositions; and using a wavelength between 600 nm or about 600 nm and 850 nm or about 850 nm, and 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 The conjugate is activated by irradiating a target area within the individual with a dose of at or about 2 J / cm of fiber length to at or about 500 J / cm of fiber length; thereby, the growth, volume or size of the tumor or lesion is reduced or inhibited.

[0041] Also provided is a method of treating a tumor or lesion. In some of any of the embodiments, the method involves administering any of the provided conjugates or any of the provided compositions to an individual having a tumor or lesion that is low-responsive, unresponsive, resistant, refractory to, or unresponsive to a previous immunotherapy, or that has recurred after a previous immunotherapy; and administering the drug at a wavelength of 600 nm or about 600 nm to 850 nm or about 850 nm at a dose of 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length to irradiate a target area where the tumor or lesion is located; wherein the method causes killing of PD-L1-expressing cells in the target area.

[0042] In some of any of the embodiments, the prior immunotherapy was treatment with an immune checkpoint inhibitor.

[0043] In some of any of the embodiments, the individual has primary or acquired resistance to a prior immunotherapy comprising PD-1 / PD-L1 blockade therapy.

[0044] Also provided is a method of treating a tumor or lesion. In some of any embodiments, the method involves administering any of the provided conjugates or any of the provided compositions to an individual who is not being treated with or has not previously received an immune checkpoint inhibitor; and administering at a wavelength of 600 nm or about 600 nm to 850 nm or about 850 nm at 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 The method further comprises irradiating a target area of ​​the individual where the tumor or lesion is located with a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length; wherein after the irradiation, the growth, size or viability of the tumor or lesion is reduced or inhibited.

[0045] In some of any of the embodiments, the conjugate is administered to a subject to treat a first tumor or lesion, inhibit its growth and / or reduce its size; and the method inhibits, delays, or prevents the emergence, growth, or establishment of one or more second tumors or lesions distal to the first tumor or lesion.

[0046] Also provided is a method of immunizing an individual having a first tumor or lesion. In some of any of the embodiments, the method involves administering to an individual having a tumor or lesion any of the provided conjugates; and administering to an individual having a tumor or lesion at a wavelength of at or about 600 nm to at or about 850 nm at a dose of 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 The invention provides a method for irradiating a target area within a first tumor or lesion with a dose of from about 2 J / cm of fiber length to about 500 J / cm of fiber length; wherein the growth of the first tumor or lesion is inhibited and / or the size of the first tumor or lesion is reduced; and the emergence, growth or establishment of one or more second tumors or lesions distal to the treated first tumor or lesion is inhibited, delayed or prevented.

[0047] In some of any embodiments, the second tumor or lesion is a metastasis of the first tumor or lesion. In some of any embodiments, the method causes killing of cells expressing PD-L1 and / or activation of an immune cell response in the vicinity of the first tumor or lesion, thereby inhibiting, delaying, or preventing the emergence, growth, or establishment of the second tumor or lesion. In some of any embodiments, the second tumor or lesion is phenotypically and / or genotypically identical to the first tumor or lesion. In some of any embodiments, the second tumor or lesion is phenotypically and / or genotypically different from the first tumor or lesion.

[0048] In some of any embodiments, the second tumor or lesion is not derived from a metastasis of the first tumor or lesion. In some of any embodiments, the method results in killing of cells expressing PD-L1 or immune cells expressing PD-L1. In some of any embodiments, the tumor or lesion comprises tumor cells, and the tumor cells do not express an immune checkpoint protein or have reduced expression of an immune checkpoint protein. In some of any embodiments, the immune checkpoint protein is selected from PD-L1, PD-1, and CTLA-4.

[0049] In some of any embodiments, the tumor cells do not express PD-L1 in response to an inflammatory stimulus. In some of any embodiments, the inflammatory stimulus is interferon.

[0050] In some of any embodiments, the tumor cells are not specifically recognized by an anti-PD-L1 antibody. In some of any embodiments, the tumor or the lesion comprises PD-L1 negative tumor cells. In some of any embodiments, at least 40%, 50%, 60%, 70%, 80%, 90% or 95% or at least about 40%, 50%, 60%, 70%, 80%, 90% or 95% of the tumor cells in the tumor or lesion are PD-L1 negative tumor cells.

[0051] In some of any embodiments, the treatment delays regrowth of the tumor or lesion, prevents recurrence of a cancer associated with the tumor or lesion, or prolongs the duration of remission of a cancer associated with the tumor or lesion. In some of any embodiments, the inhibition of growth of the tumor or lesion and / or the killing of cells expressing PD-L1 is dependent on the presence of CD8+ T cells.

[0052] In some of any embodiments, the individual is not treated with an immune checkpoint inhibitor or has not previously received treatment with an immune checkpoint inhibitor. In some of any embodiments, the individual has previously been treated with an immune checkpoint inhibitor. In some of any embodiments, the individual has a low response, no response, is resistant, is refractory to, fails to respond to, or relapsed after prior treatment with the immune checkpoint inhibitor. In some of any embodiments, the inhibition of growth, size, or viability of the tumor or lesion resulting from performing the method is greater than the inhibition resulting from prior treatment with the immune checkpoint inhibitor.

[0053] In some of any embodiments, the immune checkpoint inhibitor is an inhibitor of PD-L1, PD-1, or CTLA-4. In some of any embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor. In some of any embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some of any embodiments, the PD-1 inhibitor is an anti-CTLA-4 antibody. In some of any embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. In some of any embodiments, the PD-L 1 inhibitor is an anti-PD-L1 antibody.

[0054] In some of any of the embodiments, the method increases the number or activity of immune cells in the tumor or lesion and / or in the microenvironment of the tumor or lesion.

[0055] In some of any embodiments, the target area includes immune cells expressing PD-L1. In some of any embodiments, the cells expressing PD-L1 are immune cells. In some of any embodiments, immune cells are monocytes, macrophages, dendritic cells (DCs) or myeloid-derived suppressor cells (MDSCs). In some of any embodiments, immune cells are selected from the following groups: monocytes, macrophages (such as M1 macrophages, M2 macrophages and / or M2 tumor-associated macrophages (M2 TAMs)), dendritic cells (DCs), tolerogenic dendritic cells (tDCs) and myeloid-derived suppressor cells (MDSCs). In some of any embodiments, immune cells are monocytes. In some of any embodiments, immune cells are macrophages. In some of any embodiments, immune cells are M1 macrophages. In some of any embodiments, immune cells are M2 macrophages. In some of any embodiments, immune cells are M2 tumor-associated macrophages (M2 TAMs). In some of any embodiments, immune cells are dendritic cells (DCs). In some of any embodiments, the immune cell is a tolerogenic dendritic cell (tDC). In some of any embodiments, the immune cell is a myeloid-derived suppressor cell (MDSC). In some of any embodiments, the immune cell is located in a tumor, a tumor microenvironment, or a lymph node.

[0056] In some of any embodiments, prior to administration of the conjugate, the individual has a tumor or lesion with a low number or content of CD8+ T cell infiltration. In some of any embodiments, after the administration and the irradiation, the number, content or activity of immune cells in the tumor or lesion or in the microenvironment of the tumor or lesion increases. In some of any embodiments, after the administration and the irradiation, the number or content of CD8+ T cell infiltration in the tumor or lesion increases. In some of any embodiments, after the administration and the irradiation, the number or content of memory T cells near the tumor or lesion increases. In some of any embodiments, the targeting molecule is an antibody, antigen-binding antibody fragment or antibody-like molecule that binds to PD-L1 or comprises an antibody, antigen-binding antibody fragment or antibody-like molecule that binds to PD-L1. In some of any embodiments, the targeting molecule is an anti-PD-L1 antibody or an antigen-binding fragment thereof or comprises an anti-PD-L1 antibody or an antigen-binding fragment thereof. In some of any embodiments, the target area is a lymph node or near a lymph node. In some of any embodiments, following the administering and the irradiating, the subject exhibits a durable response, prolonged progression-free survival, reduced chance of recurrence, and / or reduced chance of metastasis.

[0057] In some of any embodiments, the irradiation is performed between 30 minutes and 96 hours after administration of the conjugate. In some of any embodiments, the irradiation is performed 24 hours ± 4 hours after administration of the conjugate. In some of any embodiments, the target area is irradiated at a wavelength of 690 ± 40 nm. In some of any embodiments, the target area is irradiated at a wavelength of 670 ± 50 nm. In some of any embodiments, the target area is irradiated at a wavelength of 50 J / cm 2 or about 50J / cm 2 Alternatively, the irradiation may be performed at a dose of 100 J / cm of fiber length or approximately 100 J / cm of fiber length.

[0058] In some of any embodiments, the tumor, lesion, or cancer is associated with a cancer selected from the group consisting of colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell carcinoma, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, small intestine cancer, spindle cell neoplasm, liver cancer, liver cancer, peripheral nerve cancer, brain cancer, skeletal muscle cancer, smooth muscle cancer, bone cancer, adipose tissue cancer, cervical cancer, uterine cancer, genital cancer, lymphoma, and multiple myeloma.

[0059] In some of any embodiments, one or more steps of the method are repeated. In some of any embodiments, administration of the antibody or antigen-binding fragment, conjugate, or composition is repeated one or more times. In some of any embodiments, after each repeated administration of the conjugate or composition, the irradiation step is repeated.

[0060] In some of any of the embodiments, the method further involves administering an additional therapeutic agent or anti-cancer therapy.

[0061] BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1A-1F The binding of exemplary anti-PD-L1 antibodies to human PD-L1 ( Figure 1A 、 Figure 1C and Figure 1E ) and cynomolgus macaque PD-L1( Figure 1B 、 Figure 1D and Figure 1F ) combination.

[0063] Figure 1G Shown is the binding specificity of exemplary anti-PD-L1 antibody 1P4 to PD-L1 compared to other B7 ligands, as determined by ELISA. Figure 1H Shown is the binding specificity of exemplary anti-PD-L1 antibody 1P9 to PD-L1 compared to other B7 ligands, as determined by ELISA.

[0064] Figure 2A Binding of exemplary anti-PD-L1 antibodies containing wild-type Fc or effector knockout Fc regions to human PD-L1 expressed on the surface of CHO cells engineered to express human PD-L1 is shown. Figure 2B Binding of exemplary anti-PD-L1 antibodies containing wild-type Fc or effector knockout Fc regions to wild-type CHO cells is shown.

[0065] Figure 3 Binding of exemplary anti-PD-L1 antibodies to A431 cancer cells expressing PD-L1 is shown.

[0066] Figure 4A-4B Shown to contain wild-type Fc region ( Figure 4A ) or effector knockout Fc region ( Figure 4B )'s ADCC activity of exemplary anti-PD-L1 antibodies.

[0067] Figure 5A-5B Exemplary anti-PD-L1-IR700 conjugates are shown to bind to human PD-L1 ( Figure 5A ) and cynomolgus macaque PD-L1( Figure 5B ) combination.

[0068] Figure 6A Binding of exemplary anti-PD-L1-IR700 conjugates containing wild-type Fc or effector knockout Fc regions to human PD-L1 expressed on the surface of CHO cells engineered to express human PD-L1 is shown. Figure 6B Binding of exemplary anti-PD-L1-IR700 conjugates containing wild-type Fc or effector knockout Fc regions to wild-type CHO cells is shown.

[0069] Figure 7A Binding of exemplary anti-PD-L1-IR700 conjugates to A431 cancer cells is shown. Figure 7B Binding of exemplary anti-PD-L1-IR700 conjugates is shown compared to avelumab-IR700 binding to interferon-γ-stimulated A431 cells. Figure 7C Binding of exemplary anti-PD-L1-IR700 conjugates is shown compared to avelumab-IR700 binding to interferon-γ-stimulated BxPC3 cells. Figure 7D Binding of exemplary anti-PD-L1-IR700 conjugates is shown compared to avelumab-IR700 binding to CHO cells engineered to express human PD-L1 (CHO-hPD-L1 cells). Figure 7E Binding of exemplary anti-PD-L1-IR700 conjugates and avelumab-IR700 bound to wild-type CHO cells is shown.

[0070] Figure 8A Shown is photoimmunotherapy-induced killing of PD-L1-expressing A431 squamous cell carcinoma cells after irradiation with 1P4-IR700, 1P9-IR700, and avelumab-IR700 conjugates.

[0071] Figure 8B-Figure 8C Shown are the results of incubation with 1P9-IR700 or avelumab-IR700 conjugates for 1 hour ( Figure 8B ) or 24 hours ( Figure 8C ) Photoimmunotherapy-induced IFN-γ-stimulated killing of A431 cancer cells following irradiation.

[0072] Figure 8D Shown are the survival rates of human peripheral blood mononuclear cells (PBMCs) from three donors after incubation with 1P9-IR700 or avelumab-IR700 and irradiation.

[0073] Figure 8E Shown are the survival rates of M1 and M2 primary human macrophages after incubation with 1P9-IR700 and irradiation.

[0074] Figures 9A-9C The results show that the Avelumab-IR700 ( Figure 9A )、1P9-IR700( Figure 9B ) and 1P4-IR700( Figure 9C ) after photoimmunotherapy-induced killing of A431 squamous cell carcinoma cells.

[0075] Figure 10A Shown are the abilities of exemplary anti-PD-L1 antibody 1P4, control antibody avelumab, and exemplary conjugate 1P4-IR700 to block PD-1 / PD-L1 interaction.

[0076] Figure 10B Shown are the abilities of exemplary anti-PD-L1 antibody 1P9, control antibody avelumab, and exemplary conjugate 1P9-IR700 to block PD-1 / PD-L1 interaction.

[0077] Figure 11 Shown are the internalization of exemplary anti-PD-L1 conjugates 1P9-IR700 and 1P4-IR700, and the reference conjugate avelumab-IR700, on pancreatic BxPC3 cancer cells. DETAILED DESCRIPTION

[0078] Provided herein are anti-PD-L1 antibodies, anti-PD-L1 antibody fragments, conjugates, compositions, combinations, methods and uses for treating individuals with tumors, lesions or cancers, for example, by activating an immune response and / or by killing target tumor cells expressing PD-L1. Also provided herein are anti-PD-L1 antibodies, anti-PD-L1 antibody fragments, conjugates, compositions, combinations, methods and uses for enhancing systemic immunity in individuals (such as cancer). In some aspects, the cancer is an invasive cancer, an infiltrating cancer or a metastatic cancer. Also provided are anti-PD-L1 antibodies, anti-PD-L1 antibody fragments, conjugates, compositions, combinations and methods for producing an enhanced response, for example, an enhanced response to a second treatment or therapy, in an individual, for example, an individual with a cancer or tumor, such as an invasive cancer, an infiltrating cancer or a metastatic cancer.

[0079] In some aspects, the embodiments provided relate to administering an anti-PD-L1 antibody provided herein to a subject to directly treat a tumor or lesion, indirectly treat a tumor or lesion, enhance systemic immunity, enhance an immune response, or enhance a response to a second treatment or therapy.

[0080] In some aspects, the embodiments provided relate to administering a conjugate to an individual, the conjugate containing any one of the antibodies or antigen-binding fragments provided, which binds to a programmed death ligand 1 (PD-L1) coupled to a phthalocyanine dye (such as a silicon phthalocyanine dye, such as any one of the dyes described in IR700 or WO2021 / 207691). In some aspects, the embodiments provided relate to irradiating a target area, such as a target area where cells expressing PD-L1 are present or may be present. In some aspects, the irradiation causes the death of cells expressing PD-L1 on the surface. In some aspects, the irradiation causes the death of tumor cells expressing PD-L1 on the surface. In some aspects, the irradiation causes the death of bone marrow cells expressing PD-L1 on the surface. In some aspects, the irradiation causes the killing of tumor cells and bone marrow cells expressing PD-L1 on the surface.

[0081] In some of any of the embodiments provided herein, provided antibodies, antibody fragments, conjugates, compositions, combinations, methods, and uses are for treating an individual having a tumor, lesion (e.g., a cancerous lesion), or cancer that is poorly responsive or substantially unresponsive to, has failed, relapsed after, is refractory to, and / or is resistant to a previous therapeutic treatment, such as a previous immunomodulatory agent treatment and / or a previous anti-cancer therapeutic treatment.

[0082] In some aspects, anti-PD-L1 antibodies and (in some cases) additional therapeutic agents are used in the compositions, combinations, methods and uses provided. In some aspects, a phthalocyanine dye-targeting molecule conjugate (e.g., a conjugate of anti-PD-L1 and IR700) is used, and in some cases, an additional therapeutic agent is used in the compositions, combinations, methods and uses provided. Uses include the use of antibodies, antibody fragments, conjugates, compositions and combinations in such methods, such as treatment methods and treatments, such as treatment regimens, and the use of such antibodies, antibody fragments, conjugates, compositions and combinations in the preparation of medicaments for the purpose of performing such treatment methods and treatments. Such antibodies, antibody fragments, conjugates, compositions and combinations for the treatment of tumors, lesions or cancers are also provided. In some aspects, such uses include performing a method or treatment as described herein, such as any treatment method or treatment regimen. In some embodiments, the methods and uses also involve irradiating a target area with light, such as the light described herein, such as a target area in which a tumor, lesion or cancer is located in the individual's body. In some embodiments, the methods and uses thereby treat the tumor, lesion or cancer. In some aspects, the tumor, lesion, or cancer to be treated includes, for example, a cancer in a subject, including a primary tumor and secondary or metastatic tumor cells, such as a secondary or metastatic cancer. In some aspects, the tumor, lesion, or cancer may include a primary tumor or multiple primary tumors and metastatic tumor cells. In some cases, the subject being treated may have one or more of a primary tumor, metastatic tumor cells, and / or invasive tumor cells.

[0083] In some aspects, methods and uses of such antibodies, antibody fragments, conjugates, compositions and combinations for enhancing, activating, inducing, eliciting, strengthening or supporting the immune function of the individual, such as local and / or systemic immunity, are also provided. In some aspects, the embodiments provided can target tumor cells. In some aspects, the embodiments provided can target cells in the tumor microenvironment, including non-cancerous cells and / or immune cells, such as antigen presenting cells or bone marrow cells with immunosuppressive functions.

[0084] One challenge in treating cancer patients is the lack of responsiveness of the cancer to therapeutic agents. Compositions and methods for treating such cancers are urgently needed. In some cases, the embodiments provided are based on the observation that treatment with the anti-PD-L1 antibodies or antibody fragments described herein results in a significant reduction in the PD-1:PD-L1 interaction. Such blocking of the PD1:PD-L1 interaction can result in substantial inhibition of tumor growth and / or a complete response to treatment with the antibody or antibody fragment. In some aspects, it was observed that the exemplary anti-PD-L1 antibodies or antibody fragments provided herein bind to PD-L1 expressed on the surface of target cells at significantly lower concentrations compared to the reference antibody, supporting the advantages of the provided antibodies or antibody fragments, such as the ability to use lower concentrations of antibodies and reduce inhibition by blocking the PD-1:PD-L1 interaction.

[0085] In some aspects, the anti-PD-L1 antibodies or antibody fragments provided also provide the advantage of retaining binding affinity after coupling with additional reagents such as phthalocyanine dyes. In contrast, it was observed that the conjugates containing the same reagent (e.g., phthalocyanine dye) and the reference anti-PD-L1 antibody exhibited a significant decrease in binding affinity, for example, a decrease of about 50 times. Such effects of retaining binding affinity after binding of the provided antibodies are unexpected and advantageous. In some aspects, the provided antibodies will retain their activity, including binding affinity, even when coupled with additional reagents, supporting the utility and advantages of antibodies for generating conjugates (such as antibody-drug conjugates and phthalocyanine dye-antibody conjugates), and the uses of the conjugates, including therapeutic uses, experimental uses, or imaging. The results described herein also indicate the advantages of the provided antibodies and antibody fragments and conjugates containing the provided antibodies and antibody fragments, such as antibody-phthalocyanine dye conjugates, in therapeutic applications, such as in photoimmunotherapy (PIT).

[0086] In some cases, embodiments are provided based on the observation that treatment of the target area with a phthalocyanine dye-targeting molecule conjugate, such as a conjugate containing an anti-PD-L1 antibody or antibody fragment described herein and a phthalocyanine dye (e.g., IR700), followed by photoirradiation (also known as "photoimmunotherapy" and "PIT"), results in significant inhibition of tumor growth and / or a complete response to treatment.

[0087] In some aspects, treatment with the anti-PD-L1 antibodies provided herein can activate, induce, enhance, or potentiate an immune response, for example, by helping to eliminate or reduce PD-L1:PD-1 interactions, which are expressed by binding to PD-L1 on immunosuppressive cells, such as immunosuppressive myeloid cells (e.g., myeloid-derived suppressor cells (MDSC), tolerogenic dendritic cells (tDC), M1 macrophages, M2 tumor-associated macrophages (M2 TAMs)). In some aspects, treatment with a phthalocyanine dye-anti-PD-L1 antibody (or antibody fragment) conjugate and light irradiation can activate, induce, enhance, or potentiate an immune response, for example, by eliminating immunosuppressive cells, such as immunosuppressive myeloid cells (e.g., myeloid-derived suppressor cells (MDSC), tolerogenic dendritic cells (tDC), M1 macrophages, M2 tumor-associated macrophages (M2 TAMs)). In some aspects, elimination of immunosuppressive cells results in activation, induction, enhancement, or potentiation of an immune response, such as an anti-tumor or anti-cancer immune response. In some aspects, any of the provided embodiments provide an advantage in that it can be used for many different tumors, lesions, or cancer types, such as cancer types of different origin or expressing different surface antigens, or cancers that may share similar immunosuppressive mechanisms. In some aspects, any of the provided embodiments can be used to overcome such immunosuppressive mechanisms.

[0088] In addition, in some aspects, the embodiments provided can provide effective treatment for heterogeneous tumors, lesions or cancers, such as tumors, lesions or cancers containing various different types of tumor or cancer cells. In some aspects, the embodiments provided also provide the advantage of inducing, activating or enhancing the local and / or systemic immune activity or systemic immunity of the individual, thereby allowing the treatment of tumors, lesions or cancers that exist elsewhere in the body rather than in the target area for irradiation, such as metastatic tumors or cancers, invasive tumors or cancers, tumors or cancers at different locations, or different types of tumors, lesions or cancers. Other benefits include treating metastatic cancers and / or invasive cancers without the need to locate and / or directly irradiate metastatic tumor cells.

[0089] The provided embodiments may also be used to treat tumors, lesions, or cancers that have not responded to previous therapeutic treatments, such as immune checkpoint inhibitors, anticancer agents, or molecules that target immunosuppressive cells. The provided embodiments also provide other advantages in cancer treatment, such as effectively treating cancers that have not responded to previous therapeutic treatments, including other anti-PD-L1 therapies.

[0090] The present disclosure also provides unexpected features in enhancing an individual's anti-cancer or anti-tumor immunity, such as anti-cancer or anti-tumor immunity against different tumors or cancers that may develop. In some cases, the embodiments provided are based on the observation that treating cancer with a phthalocyanine dye-anti-PD-L1 antibody or antibody fragment conjugate, such as an anti-PD-L1 antibody-IR700 conjugate, followed by irradiation of the tumor results in not only treatment of that particular tumor, but also effective treatment of subsequently developed tumors of the same or different types. The embodiments provided also provide for effective treatment of tumors introduced after the individual has had a complete response after the initial tumor treatment, indicating an immune memory response; and / or effective treatment of tumors distal to the target area for irradiation (e.g., metastatic tumors or tumors present in different locations). The compositions, combinations, methods and uses provided can enhance or improve an individual's immune response, such as a systemic immune response against cancer, including an immune memory response that can be effective against tumors that may develop after treatment.

[0091] In some aspects, methods involving administering an additional therapeutic agent, such as an immunomodulator, in combination with an anti-PD-L1 antibody or fragment thereof or a phthalocyanine dye conjugate (e.g., an anti-PD-L1-IR700 conjugate) are also provided.

[0092] In some of any of the embodiments provided, treatment with or administration of an anti-PD-L1 conjugate provided herein is generally followed by irradiation with a suitable wavelength of light. Such irradiation is considered part of the treatment and administration of the anti-PD-L1 conjugate provided herein, unless it is specifically stated that the irradiation step is not performed in this method. In some cases, such irradiation is referred to as photoimmunotherapy (PIT). In some cases, it is observed that the exemplary anti-PD-L1 antibodies, antibody fragments, or conjugates provided herein exhibit increased or decreased internalization compared to unconjugated antibodies or antigen-binding fragments, or conjugates comprising a reference antibody. In some cases, it is observed that the exemplary PD-L1 antibodies, antibody fragments, or conjugates provided herein exhibit increased internalization compared to unconjugated antibodies or antigen-binding fragments, or conjugates comprising a reference antibody.

[0093] All publications (including patent documents, scientific papers, and databases) mentioned in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each publication were individually incorporated by reference. If a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in patents, applications, published applications, and other publications incorporated herein by reference, the definition set forth herein controls and not the definition incorporated herein by reference.

[0094] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0095] I. PD-L1 Binding Molecules and Encoding Polynucleotides

[0096] In some aspects, PD-L1 binding molecules, such as PD-L1 binding polypeptides, are provided. Such binding molecules include antibodies (including antigen-binding fragments) that specifically bind to PD-L1 proteins, such as human PD-L1 protein (huPD-L1). Also included among the binding molecules are polypeptides containing such antibodies, including multispecific antibodies that also bind to PD-L1 in addition to one or more other antigens, such as bispecific antibodies.

[0097] PD-L1 is a ligand for the immune checkpoint protein programmed cell death 1 (PD-1) expressed in B cells, NK cells, and T cells (Shinohara et al., 1995, Genomics 23:704-6; Blank et al., 2007, Cancer Immunol Immunother 56:739-45; Finger et al., 1997, Gene 197:177-87; Pardoll, 2012, Nature Reviews Cancer 12:252-264). Under IFNγ stimulation, PD-L1 expression can be induced in dendritic cells and keratinocytes. PD-L1 is expressed on activated T cells, B cells, bone marrow cells, macrophages, and certain types of tumor cells. PD-L1 is expressed on certain immune cells, such as antigen-presenting cells (APCs), monocytes, dendritic cells (DCs), macrophages (such as M1 macrophages, M2 macrophages, M2 tumor-associated macrophages (M2TAMs)), tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs), or certain tumor cells, to induce immunosuppression near tumors or the tumor microenvironment (TME).

[0098] PD-L1 is the homologous ligand of PD-1. The complex of PD-1 and PD-L1 inhibits the proliferation of CD8+ T cells and reduces the immune response (Topalian et al., 2012, N Engl J Med 366:2443-54; Brahmer et al., 2012, N Engl J Med 366:2455-65). The main role of PD-1 is to limit the activity of T cells in peripheral tissues during inflammation in response to infection, and to limit autoimmunity (Pardoll, 2012, Nature Reviews Cancer 12:252-264). PD-1 expression is induced in activated T cells and the binding of PD-1 to one of its endogenous ligands, such as PD-L1, is used to inhibit T cell activation by inhibiting stimulatory kinases (Pardoll, 2012, Nature Reviews Cancer 12:252-264). PD-1 is also used to inhibit TCR "stop signal" (Pardoll, 2012, Nature Reviews Cancer 12: 252-264). PD-1 is highly expressed on regulatory T (Treg) cells and can increase their proliferation in the presence of ligands (Pardoll, 2012, Nature Reviews Cancer 12: 252-264). The binding of PD-L1 to PD-1 is based on the interaction with phosphatases (SHP-1 or SHP-2) via the immunoreceptor tyrosine-based switch motif (ITSM) to transmit inhibitory signals.

[0099] Therefore, the PD-L1:PD-1 pathway acts as a checkpoint in the negative regulation of some immune responses and is a key immunosuppressive mediator of T cell exhaustion. Blocking this pathway can lead to T cell activation, expansion, and enhanced effector function (Sakuishi et al., JEM Vol. 207, Sep. 27, 2010, pp. 2187-2194).

[0100] PD-L1 is also expressed on tumor cells. Tumors known to overexpress PD-L1 include, but are not limited to, those that lead to breast cancer, lung cancer, bladder cancer, colorectal cancer, head and neck cancer, various myelomas, and other cancers (Iwai et al. PNAS 99:12293-7 (2002); Ohigashi et al. Clin Cancer Res 11:2947-53 (2005)). Anti-PD-L1 antibodies have been used to treat cancers such as non-small cell lung cancer, melanoma, colorectal cancer, renal cell carcinoma, pancreatic cancer, gastric cancer, ovarian cancer, breast cancer, and hematologic malignancies (Brahmer et al., N Eng J Med 366:2455-65; Ott et al., 2013, Clin Cancer Res 19:5300-9; Radvanyi et al., 2013, Clin Cancer Res 19:5541; Menzies and Long, 2013, Ther Adv Med Oncol 5:278-85; Berger et al., 2008, Clin Cancer Res 14:13044-51). In some aspects, the use of anti-PD-L1 antibodies can reduce some of the immunosuppressive effects of PD-1 / PD-L1 by blocking the binding of PD-L1 to PD-1.

[0101] Human PD-L1 is expressed as a type I membrane precursor protein of 290 amino acids (aa) with a putative 18-aa signal peptide, a 221-aa extracellular domain, a 21-aa transmembrane region, and a 31-aa cytoplasmic domain. Antibodies and antigen-binding fragments thereof that bind, such as specifically bind to PD-L1, such as human PD-L1, and conjugates and molecules comprising such antibodies or antigen-binding fragments are provided.

[0102] A. Anti-PD-L1 Antibodies

[0103] Anti-PD-L1 antibodies are provided, including functional antigen-binding fragments. In some embodiments, the antibody or antigen-binding fragment contains a heavy chain variable region (VH) and a light chain variable region (VL), which together can specifically bind to PD-L1. In some embodiments, the antibody or antigen-binding fragment includes a full-length IgG molecule or fragment, such as scFv, Fab, or F(ab)2, that specifically binds to PD-L1, for example, human PD-L1. Among the anti-PD-L1 antibodies provided are human antibodies. Antibodies include isolated antibodies.

[0104] The term "antibody" is used herein in the broadest sense and includes polyclonal and monoclonal antibodies, including whole antibodies and functional (antigen-binding) antibody fragments, including fragment antigen-binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (VH) regions capable of specific binding to antigen, single-chain antibody fragments, including single-chain variable fragments (scFv) and single-domain antibodies (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific (e.g., bispecific or trispecific) antibodies, diabodies, triabodies and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term "antibody" should be understood to encompass functional antibody fragments thereof, also referred to herein as "antigen-binding fragments." The term also encompasses intact or full-length antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0105] The terms "complementarity determining region" and "CDR", which are synonymous with "hypervariable region" or "HVR", are known in the art to refer to non-contiguous amino acid sequences within the variable region of an antibody that confer antigen specificity and / or binding affinity. Generally, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region, and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. "Framework region" and "FR" are known in the art to refer to the non-CDR portions of the variable regions of heavy and light chains. Generally, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region, and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region.

[0106] The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those by Kabat et al., (1991), "Sequences of Proteins of Immunological Interest," 5th ed. National Institutes of Health, Public Health, Bethesda, MD. ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding sitetopography," J. Mol. Biol. 262, 732-745. ("Contact" numbering scheme); Lefranc et al., (1998) JMB 273, 927-948 ("Chothia" numbering scheme); MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Igsuperfamily V-like domains," Dev Comp Immunol, 2003 Jan;27(1):55-77 ("IMGT" numbering scheme); Honegger A and Plückthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8;309(3):657-70, ("Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272, ("AbM" numbering scheme).

[0107] The boundaries of a given CDR or FR can vary depending on the scheme used for visual identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. The numbering of both the Kabat and Chothia schemes is based on the most common antibody region sequence length, wherein insertions and deletions represented by inserted letters (e.g., "30a") occur in some antibodies. The two schemes place certain insertions and deletions ("indels") in different positions, resulting in different numbering. The Contact scheme is based on the analysis of complex crystal structures and is similar to the Chothia numbering scheme in many aspects. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on the scheme utilized by Oxford Molecular's AbM antibody modeling software.

[0108] Table 1 below lists exemplary positional boundaries for CDR-L1, CDR-L2, CDR-L3 and CDR-H1, CDR-H2, CDR-H3, as identified by the Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, the residue numbers are listed using both the Kabat and Chothia numbering schemes. FRs are located between the CDRs, e.g., where FR-L1 precedes CDR-L1, FR-L2 is between CDR-L1 and CDR-L2, FR-L3 is between CDR-L2 and CDR-L3, and so on. It should be noted that because the Kabat numbering scheme shown allows insertions at H35A and H35B, the ends of the Chothia CDR-H1 loop vary between H32 and H34 when numbered using the Kabat numbering convention shown, depending on the loop length.

[0109]

[0110]

[0111] 1-Kabat et al. (1991), "Sequences of Proteins of Immunological Interest"

[0112] Immunological Interest), 5th edition, National Institutes of Health, Public Health, Bethesda, Maryland

[0113] 2-Al-Lazikani et al., (1997) JMB 273, 927-948

[0114] Thus, unless otherwise specified, a "CDR" or "complementarity determining region" of a given antibody or region thereof (such as a variable region thereof), or a single designated CDR (e.g., CDR-H1, CDR-H2, CDR-H3), should be understood to encompass complementarity determining regions (or specific complementarity determining regions) as defined via any of the aforementioned schemes or other known schemes. For example, when stating that a particular CDR (e.g., CDR-H3) contains a given V H or V L In some embodiments, when an antibody or antigen-binding fragment thereof is stated to comprise CDR-H1, CDR-H2, and CDR-H3 as contained within a given VH region amino acid sequence and CDR-L1, CDR-L2, and CDR-L3 as contained within a given VL region amino acid sequence, the CDRs may be defined by any of the aforementioned schemes, such as the Kabat, Chothia, AbM, IMGT, or Contact methods, or other known schemes. In some embodiments, specific CDR sequences are described. Exemplary CDR sequences of the provided antibodies are described using various numbering schemes (see, for example, Tables E1a and E1b), but it is understood that the provided antibodies may include CDRs described according to any of the other aforementioned numbering schemes or other numbering schemes known to those skilled in the art.

[0115] Similarly, unless otherwise indicated, the FR or a single designated FR (e.g., FR-H1, FR-H2, FR-H3, FR-H4, FR-L1, FR-L2, FR-L3 and / or FR-L4) of a given antibody or region thereof (such as its variable region) should be understood to encompass framework regions (or specific framework regions) as defined by any known scheme. In some cases, a scheme for identifying a specific CDR, FR, or multiple FRs or CDRs is specified, such as CDRs defined by the Kabat, Chothia, AbM, IMGT, or Contact methods or other known schemes. In other cases, a specific amino acid sequence of a CDR or FR is given. In some embodiments, when an antibody or antigen-binding fragment thereof is stated to comprise FR-H1, FR-H2, FR-H3, and FR-H4 as contained within a given VH region amino acid sequence, and FR-L1, FR-L2, FR-L3, and FR-L4 contained within a given VL region amino acid sequence, the CDRs can be defined by any of the aforementioned schemes, such as the Kabat, Chothia, AbM, IMGT, or Contact methods, or other known schemes.

[0116] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain involved in binding the antibody to an antigen. The variable regions of the heavy and light chains of a native antibody (VH and VL, respectively) generally have a similar structure, with each domain comprising four conserved framework regions (FRs) and three CDRs. (See, e.g., Kindt et al., Kuby Immunology, 6th ed., WH Freeman and Co., p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be isolated using a VH domain or VL domain from an antibody that binds the antigen to screen a library of complementary VL domains or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0117] An antibody fragment is a fragment of a provided antibody. An "antibody fragment" or "antigen-binding fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; heavy chain variable (V H ) region, single-chain antibody molecules such as scFv and those containing only V H In a specific embodiment, the antibody is a single domain antibody comprising a variable heavy chain (V H ) and variable light chain (V L ) region or an antibody fragment containing a variable heavy chain (V H ) and variable light chain (V L In some embodiments, the antibody is an antibody fragment comprising a heavy chain variable (V H ) region and / or light chain variable (V L ) region, such as a single-chain antibody fragment, such as scFv.

[0118] Antibody fragments can be produced by various techniques, including but not limited to proteolytic digestion of intact antibodies and production by recombinant host cells. In some embodiments, antibodies are recombinantly produced fragments, such as fragments comprising arrangements that do not occur naturally, such as fragments having two or more antibody regions or chains joined by synthetic linkers (e.g., peptide linkers), and / or fragments that cannot be produced by enzymatic digestion of naturally occurring intact antibodies. In some aspects, the antibody fragment is a scFv.

[0119] "Humanized" antibodies are antibodies in which all or substantially all of the CDR amino acid residues are derived from non-human CDRs and all or substantially all of the FR amino acid residues are derived from human FRs. Humanized antibodies optionally may include at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of a non-human antibody refers to a non-human antibody variant that has undergone humanization (generally to reduce immunogenicity to humans) while retaining the specificity and affinity of the parent non-human antibody. In some embodiments, some FR residues in the humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., an antibody from which the CDR residues were derived), e.g., to restore or improve antibody specificity or affinity.

[0120] Among the anti-PD-L1 antibodies provided are human antibodies. A "human antibody" is an antibody of non-human origin that has an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human or human cell, or has a sequence that uses a human antibody repertoire or encodes other human antibodies (including human antibody libraries). The term does not include humanized forms of non-human antibodies that contain non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human CDRs. The term includes antigen-binding fragments of human antibodies.

[0121] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce complete human antibodies or complete antibodies with human variable regions in response to an antigenic challenge. Such animals typically contain all or part of a human immunoglobulin locus, which replaces the endogenous immunoglobulin locus, or is present outside the chromosome or randomly integrated into the animal chromosome. In such transgenic animals, the endogenous immunoglobulin locus is generally no longer activated. Human antibodies can also be derived from human antibody libraries, including phage display libraries and cell-free libraries, which contain sequences encoding antibodies derived from the human repertoire.

[0122] Among the antibodies provided are monoclonal antibodies, including monoclonal antibody fragments. As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population or obtained within the population, that is, except for the possible variants containing naturally occurring mutations or occurring during the production of monoclonal antibody preparations, the individual antibodies constituting the population are identical, and such variants generally exist in small quantities. Compared with polyclonal antibody preparations that generally include different antibodies for different epitopes, each monoclonal antibody of a monoclonal antibody preparation is for a single epitope on an antigen. The term should not be interpreted as requiring the production of antibodies via any ad hoc method. Monoclonal antibodies can be obtained via a variety of techniques, including but not limited to self-hybridoma production, recombinant DNA methods, phage display and other antibody presentation methods.

[0123] The terms "polypeptide" and "protein" are used interchangeably and refer to a polymer of amino acid residues and are not limited to a minimum length. Polypeptides (including the antibodies and antibody conjugates provided) may include amino acid residues, including natural and / or non-natural amino acid residues. The term also includes post-translational modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, and the like. In some aspects, the polypeptide may contain modifications relative to the native or natural sequence, as long as the protein maintains the desired activity. These modifications may be intentional, such as through site-directed mutagenesis, or may be accidental, such as through mutations in the host producing the protein or errors caused by PCR amplification.

[0124] 1. Exemplary Antibodies

[0125] In some embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof contains a heavy chain variable region (VH) and a light chain variable region (VL) as described herein, or a sufficient antigen-binding portion thereof. In some embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof contains a VH region sequence, or a sufficient antigen-binding portion thereof, containing CDR-H1, CDR-H2, and / or CDR-H3 as described herein. In some embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof contains a VL region sequence, or a sufficient antigen-binding portion thereof, containing CDR-L1, CDR-L2, and / or CDR-L3 as described herein. In some embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof contains a VH region sequence, containing CDR-H1, CDR-H2, and / or CDR-H3 as described, and a VL region sequence, containing CDR-L1, CDR-L2, and / or CDR-L3 as described herein. Also included among the antibodies provided are those having sequences that are at least or about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to such sequences.

[0126] In some embodiments, the antibody or antigen-binding fragment thereof has a heavy chain variable (VH) region having a sequence selected from any one of SEQ ID NOs: 1-16 and 330-334, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a VH region amino acid selected from any one of SEQ ID NOs: 1-16 and 330-334, or contains CDR-H1, CDR-H2 and / or CDR-H3 present in such a VH sequence.

[0127] In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any one of SEQ ID NOs: 1-11, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with a VH region amino acid selected from any one of SEQ ID NOs: 1-11, or containing CDR-H1, CDR-H2 and / or CDR-H3 present in such a VH sequence.

[0128] In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any one of SEQ ID NOs: 12-16, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a VH region amino acid selected from any one of SEQ ID NOs: 12-16, or containing CDR-H1, CDR-H2 and / or CDR-H3 present in such a VH sequence.

[0129] In some embodiments, the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment having a VH region comprising CDR-H1, CDR-H2, and / or CDR-H3 from a VH region selected from any one of SEQ ID NOs: 12 to 16. In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any one of SEQ ID NOs: 330 to 334, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region amino acid selected from any one of SEQ ID NOs: 330 to 334, or containing CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence.

[0130] In some embodiments, the antibody or antigen-binding fragment thereof has a VH region having a sequence selected from any one of SEQ ID NOs: 1, 2, 6, and 7, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with a VH region amino acid selected from any one of SEQ ID NOs: 1, 2, 6, and 7, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VH region set forth in SEQ ID NO: 1, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with a VH region amino acid selected from any one of SEQ ID NOs: 1, 2, 6, and 7, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VH region set forth in SEQ ID NO: 2, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VH region set forth in SEQ ID NO: 6, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VH region set forth in SEQ ID NO:7, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:7, or contains CDR-H1, CDR-H2 and / or CDR-H3 present in such a VH sequence.

[0131] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any one of SEQ ID NOs: 1-16. In some of these embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Chothia. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to AbM. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Kabat. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Contact. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to IMGT.

[0132] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any one of SEQ ID NOs: 1-11. In some of these embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Chothia. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to AbM. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Kabat. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Contact. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to IMGT.

[0133] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any one of SEQ ID NOs: 12-16. In some of these embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Chothia. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to AbM. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Kabat. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Contact. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to IMGT.

[0134] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any one of SEQ ID NOs: 330-334. In some of these embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Chothia. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to AbM. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Kabat. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to Contact. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and / or CDR-H3 numbered according to IMGT.

[0135] In some embodiments, the VH region of an antibody or antigen-binding fragment thereof provided comprises CDR-H1, CDR-H2, and / or CDR-H3 according to Chothia numbering, as shown in Table E1a. In some embodiments, the VH region of an antibody or antigen-binding fragment thereof provided comprises CDR-H1, CDR-H2, and / or CDR-H3 according to AbM numbering, as shown in Table E1a. In some embodiments, the VH region of an antibody or antigen-binding fragment thereof provided comprises CDR-H1, CDR-H2, and / or CDR-H3 according to Kabat numbering, as shown in Table E1a. In some embodiments, the VH region of an antibody or antigen-binding fragment thereof provided comprises CDR-H1, CDR-H2, and / or CDR-H3 according to Contact numbering, as shown in Table E1a. In some embodiments, the VH region of an antibody or antigen-binding fragment thereof provided comprises CDR-H1, CDR-H2, and / or CDR-H3 according to IMGT numbering, as shown in Table E1a.

[0136] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VH region comprising a CDR-H1, CDR-H2, and CDR-H3 selected from the following groups: CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 35, 36, and 37, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 48, 49, and 37, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 58, 59, and 60, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 71, 72, and 60, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 82, 83, and 84, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 83, 84, and 85, respectively; NO: 48, 95 and 84; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 48, 104 and 105, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 116, 117, 118, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 129, 130 and 131, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 142, 143 and 144, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 155, 156 and 157, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 157, 158 and 159, respectively; NO: 168, 169 and 170; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 35, 181 and 182, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 168, 169 and 193, respectively; CDR-H1, CDR-H2 and CDR-H3 comprising the sequences of SEQ ID NO: 197, 198 and 199, respectively; numbered according to Chothia.

[0137] For example, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising a CDR-H1, CDR-H2, and CDR-H3 comprising sequences selected from the group consisting of SEQ ID NOs: 35, 36, and 37; SEQ ID NOs: 48, 49, and 37; SEQ ID NOs: 58, 59, and 60; SEQ ID NOs: 71, 72, and 60; SEQ ID NOs: 82, 83, and 84; SEQ ID NOs: 48, 95, and 84; SEQ ID NOs: 48, 104, and 105; SEQ ID NOs: 116, 117, 118; SEQ ID NOs: 129, 130, and 131; and SEQ ID NOs: 142, 143, and 144 (according to Chothia numbering).

[0138] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising a CDR-H1, CDR-H2, and CDR-H3 comprising a sequence selected from the group consisting of SEQ ID NOs: 35, 36, and 37; SEQ ID NOs: 48, 49, and 37; SEQ ID NOs: 82, 83, and 84; SEQ ID NOs: 48, 95, and 84 (numbered according to Chothia). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising a CDR-H1, CDR-H2, and CDR-H3 comprising a sequence of SEQ ID NOs: 35, 36, and 37, respectively (numbered according to Chothia). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising a CDR-H1, CDR-H2, and CDR-H3 comprising a sequence of SEQ ID NOs: 82, 83, and 84, respectively (numbered according to Chothia).

[0139] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 (according to Kabat numbering) comprising the sequences of SEQ ID NOs: 40, 41, and 37, respectively. In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 (according to Kabat numbering) comprising the sequences of SEQ ID NOs: 87, 88, and 84, respectively.

[0140] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any one of SEQ ID NOs: 1-16. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any one of SEQ ID NOs: 1-11. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any one of SEQ ID NOs: 12-16. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any one of SEQ ID NOs: 330-334. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence of SEQ ID NO: 1. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence of SEQ ID NO: 2. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the VH region sequence of SEQ ID NO: 6. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the VH region sequence of SEQ ID NO: 7.

[0141] Also provided are antibodies and antigen-binding fragments thereof having sequences that are at least or about at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to such sequences. For example, provided herein is an antibody or antigen-binding fragment comprising a VH region comprising a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a VH region sequence selected from any one of SEQ ID NOs: 1-16 and 330-334.

[0142] In some embodiments, the antibody comprises only V H A single domain antibody comprising a V domain sequence or a sufficient antigen binding portion thereof, such as the V domain sequence described above H Any one of the sequences (e.g., CDR-H1, CDR-H2 and / or CDR-H3).

[0143] In some embodiments, the antibodies (e.g., anti-PD-L1 antibodies) or antigen-binding fragments thereof provided herein comprising a VH region further comprise a light chain or sufficient antigen-binding portion thereof. For example, in some embodiments, the antibody or antigen-binding fragment thereof contains a VH region and a VL region, or sufficient antigen-binding portions of a VH and VL region. In such embodiments, the VH region sequence may be any of the VH sequences described above. In some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or scFv. In some such embodiments, the antibody is a full-length antibody also containing a constant region.

[0144] In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any one of SEQ ID NOs: 17-34 and 335-340, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 17-34 and 335-340, or containing the CDR-L1, CDR-L2 and / or CDR-L3 present in such VL sequences.

[0145] In some embodiments, the antibody or antigen-binding fragment thereof has a VL having a sequence selected from any one of SEQ ID NOs: 17-28, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 17-28, or containing the CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0146] In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any one of SEQ ID NOs: 29-34, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 29-34, or containing CDR-L1, CDR-L2 and / or CDR-L3 present in such VL sequences.

[0147] In some embodiments, the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment having a VL region comprising CDR-L1, CDR-L2, and / or CDR-L3 from a VL region selected from any one of SEQ ID NOs: 29 to 34. In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any one of SEQ ID NOs: 335 to 340, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 335 to 340, or containing CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0148] In some embodiments, the antibody or antigen-binding fragment thereof has a VL having a sequence selected from any one of SEQ ID NOs: 21-24, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 21-24, or comprising a CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VL region set forth in SEQ ID NO: 21, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any one of SEQ ID NOs: 21-24, or comprising a CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VL region set forth in SEQ ID NO: 22, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 22, or contains CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VL region set forth in SEQ ID NO: 23, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 23, or contains CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence. In some embodiments, the antibody or antigen-binding fragment thereof has a VL region set forth in SEQ ID NO:24, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:24, or contains CDR-L1, CDR-L2 and / or CDR-L3 present in such a VL sequence.

[0149] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Chothia in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to AbM in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Kabat in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Contact in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to IMGT numbering in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-34.

[0150] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Chothia in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to AbM in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Kabat in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Contact in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to IMGT numbering in a VL comprising a sequence selected from any one of SEQ ID NOs: 17-28.

[0151] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Chothia in a VL comprising a sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to AbM in a VL comprising a sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Kabat in a VL comprising a sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Contact in a VL comprising a sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to IMGT numbering in a VL comprising a sequence selected from any one of SEQ ID NOs: 29-34.

[0152] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Chothia in a VL comprising a sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to AbM in a VL comprising a sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Kabat in a VL comprising a sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise CDR-L1, CDR-L2, and / or CDR-L3 numbered according to Contact in a VL comprising a sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to IMGT numbering in a VL comprising a sequence selected from any one of SEQ ID NOs: 335-340.

[0153] In some embodiments, the VL region of an antibody or antigen-binding fragment thereof is provided and comprises CDRL1, CDRL2, and / or CDR-L3 according to Chothia numbering, as shown in Table E1b. In some embodiments, the VL region of an antibody or antigen-binding fragment thereof is provided and comprises CDRL1, CDRL2, and / or CDR-L3 according to AbM numbering, as shown in Table E1b. In some embodiments, the VL region of an antibody or antigen-binding fragment thereof is provided and comprises CDRL1, CDRL2, and / or CDR-L3 according to Kabat numbering, as shown in Table E1b. In some embodiments, the VL region of an antibody or antigen-binding fragment thereof is provided and comprises CDRL1, CDRL2, and / or CDR-L3 according to Contact numbering, as shown in Table E1b. In some embodiments, the VL region of an antibody or antigen-binding fragment thereof is provided and comprises CDRL1, CDRL2, and / or CDR-L3 according to IMGT numbering, as shown in Table E1b.

[0154] In some embodiments of the antibodies or antigen-binding fragments thereof provided herein, the VL region comprises a CDR-L1, CDR-L2, and CDR-L3 selected from the group consisting of: CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 210, 211, and 212, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 218, 211, and 212, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 221, 222, and 223, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 229, 222, and 223, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 233, 234, and 235, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 236, 237, and 238, respectively; NO: 241, 234 and 242; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 246, 247 and 248, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 246, 254 and 255, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 258, 259 and 260, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 265, 266 and 267, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 246, 273 and 274, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 274, respectively; NO: 278, 279 and 280; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 286, 287 and 288, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 294, 234 and 295, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 299, 300 and 301, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 306, 234 and 307, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 311, 312 and 313, respectively; CDR-L1, CDR-L2 and CDR-L3 comprising the sequence of SEQ ID NO: 314, 315 and 316, respectively; CDR-L1, CDR-L2 and CDR-L3 of the sequences of NO:319, 320 and 321.

[0155] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising a CDR-L1, CDR-L2, and CDR-L3 comprising sequences selected from the group consisting of SEQ ID NOs: 210, 211, and 212; SEQ ID NOs: 218, 211, and 212; SEQ ID NOs: 221, 222, and 223; SEQ ID NOs: 229, 222, and 223; SEQ ID NOs: 233, 234, and 235; SEQ ID NOs: 241, 234, and 242; SEQ ID NOs: 246, 247, and 248; SEQ ID NOs: 246, 254, and 255; SEQ ID NOs: 258, 259, and 260; SEQ ID NOs: 265, 266, and 267; SEQ ID NOs: 246, 273, and 274; SEQ ID NOs: 278, 279, and 280, respectively (according to Chothia numbering). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively (according to Chothia numbering). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively (according to Chothia numbering).

[0156] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively (according to Kabat numbering). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively (according to Kabat numbering).

[0157] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VL region comprising a CDR-L1, CDR-L2, and CDR-L3 comprising a sequence selected from the group consisting of: SEQ ID NOs: 233, 234, and 235; SEQ ID NOs: 241, 234, and 242; SEQ ID NOs: 246, 247, and 248; SEQ ID NOs: 246, 254, and 255 (according to Kabat numbering, Chothia numbering, or AbM numbering).

[0158] In some embodiments, the antibodies or antigen-binding fragments thereof provided contain CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a VL region sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided contain CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a VL region sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibodies or antigen-binding fragments thereof provided contain CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a VL region sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided contain CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a VL region sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the antibodies provided contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO: 21. In some embodiments, the antibodies provided contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO: 22. In some embodiments, the antibodies provided contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO: 23. In some embodiments, the antibodies provided contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO: 24.

[0159] In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence selected from any one of SEQ ID NOs: 29-34. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence selected from any one of SEQ ID NOs: 21. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence of SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence of SEQ ID NO: 23. In some embodiments, the antibodies or antigen-binding fragments thereof provided comprise a VL region comprising a sequence of SEQ ID NO: 24. Antibodies having a sequence that is at least or about at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to any of such sequences are also provided.

[0160] In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 1-16 and the VL region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 1-11 and the VL region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 330-334 and the VL region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 1, 2, 6 and 7 and the VL region of the antibody or fragment comprises a sequence selected from any one of SEQ ID NOs: 21-24. In some embodiments, the VH region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 1, and the VL region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 21. In some embodiments, the VH region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 2, and the VL region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 22. In some embodiments, the VH region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 6, and the VL region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 23. In some embodiments, the VH region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 7, and the VL region of the antibody or fragment comprises the sequence set forth in SEQ ID NO: 24.

[0161] Also provided are antibodies and antigen-binding fragments thereof having sequences that are at least or about at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to such sequences. For example, provided herein are antibodies or antigen-binding fragments comprising a VH region comprising a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a VH region sequence selected from any one of SEQ ID NOs: 1-16, and / or comprising a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a VH region sequence selected from any one of SEQ ID NOs: 17-34. In some embodiments, the antibody or antigen-binding fragment contains a VH region comprising a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VH region sequence selected from any one of SEQ ID NOs: 1-11, and a VL region that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VL region sequence selected from any one of SEQ ID NOs: 17-28. In some embodiments, the antibody or antigen-binding fragment contains a VH region comprising a sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VH region sequence selected from any one of SEQ ID NOs: 330-334, and a VL region that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VL region sequence selected from any one of SEQ ID NOs: 335-340. In some embodiments, the antibody or antigen-binding fragment contains a VH region comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region sequence selected from any one of SEQ ID NOs: 1, 2, 6, and 7, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region sequence selected from any one of SEQ ID NOs: 21-24.In some embodiments, the antibody or antigen-binding fragment comprises a VH region comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 1, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 21. In some embodiments, the antibody or antigen-binding fragment comprises a VH region comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:2, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:22. In some embodiments, the antibody or antigen-binding fragment comprises a VH region comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:6, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:23. In some embodiments, the antibody or antigen-binding fragment comprises a VH region comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:7, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO:24.

[0162] In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to a VH sequence selected from any one of SEQ ID NOs: 1-16; and a VL having at least 90% sequence identity to a VL sequence selected from any one of SEQ ID NOs: 17-34.

[0163] In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to a VH sequence selected from any one of SEQ ID NOs: 1, 2, 6, and 7; and a VL having at least 90% sequence identity to a VL sequence selected from any one of SEQ ID NOs: 21-24. In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to a VH sequence of SEQ ID NO: 1; and a VL having at least 90% sequence identity to a VL sequence of SEQ ID NO: 21. In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to a VH sequence of SEQ ID NO: 2; and a VL having at least 90% sequence identity to a VL sequence of SEQ ID NO: 22. In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to a VH sequence of SEQ ID NO: 6; and a VL having at least 90% sequence identity to a VL sequence of SEQ ID NO: 23. In some embodiments, the antibody or antigen-binding fragment comprises a VH having at least 90% sequence identity to the VH sequence of SEQ ID NO:7; and a VL having at least 90% sequence identity to the VL sequence of SEQ ID NO:24.

[0164] In some embodiments, the VH region of the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence of any one selected from SEQ ID NOs: 1-16; and the VL region of the antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence of any one selected from SEQ ID NOs: 17-34.

[0165] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence of any one selected from SEQ ID NOs: 1-11, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence of any one selected from SEQ ID NOs: 17-28.

[0166] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence of any one selected from SEQ ID NOs: 12-16, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence of any one selected from SEQ ID NOs: 29-34.

[0167] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the VH region sequence of any one selected from SEQ ID NOs: 330-334, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the VL region sequence of any one selected from SEQ ID NOs: 335-340.

[0168] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained within the VH region sequence of any one selected from SEQ ID NOs: 1, 2, 6, and 7, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained within the VL region sequence of any one selected from SEQ ID NOs: 21-24.

[0169] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 1, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 17. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:2, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:18. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:3, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:19. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:4, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:19. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:5, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:20.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:1, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:21. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:2, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:22. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:6, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:23. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:7, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:24. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:8, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:25.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO:9, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO:26. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 10, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 27. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 11, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 28. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 12, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 29. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 13, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 30.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 14, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 31. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 15, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 32. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 12, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 33. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, and CDR-H3, which respectively comprise the sequences of CDR-H1, CDR-H2, and CDR-H3 contained in the VH region sequence shown in SEQ ID NO: 16, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3, which respectively comprise the sequences of CDR-L1, CDR-L2, and CDR-L3 contained in the VL region sequence shown in SEQ ID NO: 34.

[0170] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH comprising CDR-H1, CDR-H2, and CDR-H3, wherein: CDR-H1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 35, 38, 40, 42, 45, 48, 50, 52, 54, 56, 58, 61, 63, 65, 68, 71, 73, 75, 77, 79, 82, 85, 87, 89, 92, 96, 98, 100, 102, 106, 108, 110, 113, 116, 119, 121, 123, 126, 129, 32, 134, 136, 139, 142, 145, 147, 149, 152, 155, 158, 160, 162, 165, 168, 171, 173, 175, 178, 183, 185, 187, 190, 197, 200, 202, 204, and 207; CDR-H2 comprises a sequence selected from SEQ ID NO:36, 39, 41, 43, 46, 49, 51, 53, 55, 57, 59, 62, 64, 66, 69, 72, 74, 76, 78, 80, 83, 86, 8 8, 90, 93, 95, 97, 99, 101, 103, 104, 107, 109, 111, 114, 117, 120, 122, 124, 127, 130, 1 33, 135, 137, 140, 143, 146, 148, 150, 153, 156, 159, 161, 163, 166, 169, 172, 174, 176, 179, 181, 184, 186, 188, 191, 194, 198, 201, 203 and 205208; and CDR-H3 comprises a sequence selected from SEQ the sequence of any one of ID NOs: 37, 44, 47, 60, 67, 70, 81, 84, 91, 94, 105, 112, 115, 118, 125, 128, 131, 138, 141, 144, 151, 154, 157, 164, 167, 170, 177, 180, 182, 189, 192, 193, 195, 196, 199, 206, and 209;and a VL comprising CDR-L1, CDR-L2, and CDR-L3, wherein CDR-L1 comprises a sequence selected from any one of SEQ ID NOs: 210, 213, 216, 218, 219, 220, 221, 224, 227, 229, 230, 232, 233, 236, 239, 241, 243, 245, 246, 249, 252, 258, 261, 264, 265, 268, 271, 278, 281, 284, 286, 289, 292, 294, 296, 298, 299, 302, 306, 308, 310, 311, 314, 317, 319, 322, and 325; ID NO: 211, 214, 217, 222, 225, 228, 231, 234, 237, 240, 247, 250, 253, 254, 256, 259, 262, 266, 269, 272, 273, 275, 277, 279, 282, 285, 287, 290, 293, 300, 303, 305, 312, 315, 318, 320, 323 and 326; and CDR-L3 comprises a sequence selected from SEQ ID NO: The sequence of any one of NO: 212, 215, 223, 226, 235, 238, 242, 244, 248, 251, 255, 257, 260, 263, 267, 270, 274, 276, 280, 283, 288, 291, 295, 297, 301, 304, 307, 309, 313, 316, 321 and 324. ;

[0171] In some embodiments, CDR-H1 comprises a sequence selected from any one of SEQ ID NOs: 35, 38, 40, 42, 45, 48, 50, 52, 54, 56, 82, 85, 87, 89, 92, 96, 98, 100, and 102; CDR-H2 comprises a sequence selected from any one of SEQ ID NOs: 36, 39, 41, 43, 46, 49, 51, 53, 55, 57, 83, 86, 88, 90, 93, 95, 97, 99, 101, and 103; and CDR-H3 comprises a sequence selected from any one of SEQ ID NOs: 37, 44, 47, 84, 91, and 94; and CDR-L1 comprises a sequence selected from any one of SEQ ID NOs: NO: 233, 236, 239, 241, 243, 245, 246, 249 and 252; CDR-L2 comprises a sequence selected from any one of SEQ ID NO: 217, 234, 237, 240, 247, 250, 253, 254 and 256; and CDR-L3 comprises a sequence selected from any one of SEQ ID NO: 235, 238, 242, 244, 248, 251, 255 and 257. In some embodiments, CDR-H1 comprises the sequence of SEQ ID NO: 35; CDR-H2 comprises the sequence of SEQ ID NO: 36; and CDR-H3 comprises the sequence of SEQ ID NO: 37; and CDR-L1 comprises the sequence of SEQ ID NO: 233; CDR-L2 comprises the sequence of SEQ ID NO: 234; and CDR-L3 comprises the sequence of SEQ ID NO: 235. In some embodiments, CDR-H1 comprises the sequence of SEQ ID NO:82; CDR-H2 comprises the sequence of SEQ ID NO:83; and CDR-H3 comprises the sequence of SEQ ID NO:84; and CDR-L1 comprises the sequence of SEQ ID NO:246; CDR-L2 comprises the sequence of SEQ ID NO:247; and CDR-L3 comprises the sequence of SEQ ID NO:248.

[0172] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH and a VL, wherein: the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 35, 36, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 210, 211, and 212, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 48, 49, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 218, 211, and 212, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 58, 59, and 60, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 219, 220, 221, and 222, respectively. NO: 221, 222 and 223 CDR-L1, CDR-L2 and CDR-L3; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 58, 59 and 60, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 221, 222 and 223, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 71, 72 and 60, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 229, 222 and 223, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 35, 36 and 37, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 37, 38 and 39, respectively. NO: 233, 234 and 235 CDR-L1, CDR-L2 and CDR-L3; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 48, 49 and 37, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 241, 234 and 242, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 82, 83 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 246, 247 and 248, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 48, 95 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 247, CDR-L1, CDR-L2 and CDR-L3 of NOs:246, 254 and 255;The VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 48, 104 and 105, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 258, 259 and 260, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 116, 117 and 118, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 265, 266 and 267, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 129, 130 and 131, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 246, 273 and 274, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 1 NO: 142, 143 and 144 CDR-H1, CDR-H2 and CDR-H3, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 278, 279 and 280, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 155, 156 and 157, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 286, 287 and 288, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 168, 169 and 170, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 294, 234 and 295, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 168, 169 and 170, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 294, 234 and 295, respectively; NOs: 35, 181 and 182, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 299, 300 and 301, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 168, 169 and 193, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 306, 234 and 307, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 155, 156 and 157, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 311, 312 and 313, respectively;VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 197, 198, and 199, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 319, 320, and 321, respectively.;

[0173] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH and a VL, wherein: the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 35, 36, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 48, 49, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 241, 234, and 242, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 82, 83, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 242, 243, and 244, respectively. NO: 246, 247 and 248; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 48, 95 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 246, 254 and 255, respectively. In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 35, 36 and 37, respectively, and a VL region comprising CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 233, 234 and 235, respectively (according to Chothia numbering). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 82, 83, and 84, respectively, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively (according to Chothia numbering).

[0174] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 40, 41, and 37, respectively, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively (according to Kabat numbering). In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 87, 88, and 84, respectively, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively (according to Kabat numbering).

[0175] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH and a VL, wherein: the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 40, 41, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 210, 211, and 212, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 52, 53, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 218, 211, and 212, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 63, 64, and 60, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 64, 64, and 60, respectively. NO: 221, 222 and 223 CDR-L1, CDR-L2 and CDR-L3; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 63, 64 and 60, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 221, 222 and 223, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 75, 76 and 60, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 229, 222 and 223, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 40, 41 and 37, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 40, NO: 233, 234 and 235 CDR-L1, CDR-L2 and CDR-L3; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 52, 53 and 37, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 241, 234 and 242, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 87, 88 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 246, 247 and 248, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 98, 99 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 98, CDR-L1, CDR-L2 and CDR-L3 of NOs:246, 254 and 255;The VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 108, 109 and 105, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 258, 259 and 260, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 121, 122 and 118, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 265, 266 and 267, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 134, 135 and 131, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 246, 273 and 274, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 134, 135 and 131, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 246, 273 and 274, respectively. NO: 147, 148 and 144 CDR-H1, CDR-H2 and CDR-H3, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 278, 279 and 280, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 160, 161 and 157, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 286, 287 and 288, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 173, 174 and 170, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 294, 234 and 295, respectively; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NO: 173, 174 and 170, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 173, 174 and 170, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NO: 173 NOs: 185, 186 and 182, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 299, 300 and 301, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 173, 174 and 193, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 306, 234 and 307, respectively; the VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 160, 161 and 157, respectively, and the VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 311, 312 and 313, respectively;VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 202, 203, and 199, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 319, 320, and 321, respectively.;

[0176] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH and a VL, wherein: the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 40, 41, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 52, 53, and 37, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 241, 234, and 242, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 87, 88, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 242, 243, and 244, respectively. NOs: 246, 247 and 248; VH comprises CDR-H1, CDR-H2 and CDR-H3 comprising SEQ ID NOs: 98, 99 and 84, respectively, and VL comprises CDR-L1, CDR-L2 and CDR-L3 comprising SEQ ID NOs: 246, 254 and 255, respectively.

[0177] In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 1 and 17, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 2 and 18, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 3 and 19, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 4 and 19, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 5 and 20, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 1 and 21, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 2 and 22, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 6 and 23, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 7 and 24, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: ID NO: 8 and 25; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NO: 9 and 26, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NO: 10 and 27, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NO: 11 and 28, respectively.

[0178] In some embodiments, the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 13 and 30, respectively, or humanized sequences thereof; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 14 and 31, respectively, or humanized sequences thereof; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 15 and 32, respectively, or humanized sequences thereof; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 12 and 33, respectively, or humanized sequences thereof; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs: 16 and 34, respectively, or humanized sequences thereof.

[0179] In some embodiments, the antibody or antigen-binding fragment comprises one or more VH and one or more VL in any order or orientation. In some embodiments, the antibody or antigen-binding fragment comprises a VH region and a VL region, and the VH region is amino-terminal to the VL region. In some embodiments, the antibody or antigen-binding fragment comprises a VH region and a VL region, and the VH region is carboxy-terminal to the VL region. In some embodiments, the VH region and the VL region are directly or indirectly linked, optionally through a linker.

[0180] In some embodiments, an antibody or antigen-binding fragment (e.g., scFv) may comprise a VH region or a portion thereof, followed by a linker, followed by a VL region or a portion thereof. In some embodiments, an antibody or antigen-binding fragment (e.g., scFv) may comprise a VL region or a portion thereof, followed by a linker, followed by a VH region or a portion thereof.

[0181] In some embodiments, the antibody or its antigen-binding fragment is a single-chain antibody fragment, such as a single-chain variable fragment (scFv) or a diabody or a single-domain antibody (sdAb). In some embodiments, the antibody or antigen-binding fragment is a single-domain antibody comprising only a VH region. In some embodiments, the antibody or antigen-binding fragment is an scFv comprising a heavy chain variable (VH) region and a light chain variable (VL) region. In some embodiments, a single-chain antibody fragment (e.g., scFv) includes one or more linkers that join two antibody domains or regions (such as a VH region and a VL region). The linker is typically a peptide linker, such as a flexible and / or soluble peptide linker. Among the linkers are those rich in glycine and serine and / or in some cases rich in threonine. In some embodiments, the linker further includes charged residues that can improve solubility, such as lysine and / or glutamic acid. In some embodiments, the linker further includes one or more proline residues.

[0182] Thus, provided anti-PD-L1 antibodies include single-chain antibody fragments, such as scFv and diabodies, particularly human single-chain antibody fragments, which generally comprise two antibody domains or regions joined (such V H and V L The linker is typically a peptide linker, for example a flexible and / or soluble peptide linker, such as a peptide linker rich in glycine and serine.

[0183] The antibody or antigen-binding fragment thereof may contain an immunoglobulin constant region, such as at least a portion of one or more constant region domains. In some embodiments, the constant region includes a light chain constant region and / or a heavy chain constant region 1 (CH1). In some embodiments, the antibody includes a CH2 and / or CH3 domain, such as an Fc region. In some embodiments, the Fc region is that of a human IgG, such as an IgG1 or IgG4.

[0184] Also provided are nucleic acids, such as polynucleotides, encoding antibodies and / or portions thereof, such as chains thereof. Among the nucleic acids provided are nucleic acids encoding the anti-PD-L1 antibodies and fragments described herein. Also provided are nucleic acids, such as polynucleotides, encoding one or more antibodies and / or portions thereof, such as nucleic acids encoding one or more of the anti-PD-L1 antibodies or antigen-binding fragments described herein, and / or other antibodies and / or portions thereof, such as antibodies and / or portions thereof that bind to other target antigens. Nucleic acids may include nucleic acids comprising naturally and / or non-naturally occurring nucleotides and bases, including, for example, nucleic acids with backbone modifications. The terms "nucleic acid molecule," "nucleic acid," and "polynucleotide" are used interchangeably and refer to nucleotide polymers. Such nucleotide polymers may contain natural and / or non-natural nucleotides and include, but are not limited to, DNA, RNA, and PNA. A "nucleic acid sequence" refers to a linear sequence of nucleotides comprising a nucleic acid molecule or polynucleotide.

[0185] Also provided are vectors containing nucleic acids, such as polynucleotides, and host cells containing the vectors, for example, for use in producing antibodies or antigen-binding fragments thereof. Methods for producing antibodies or antigen-binding fragments thereof are also provided. The nucleic acid may encode an amino acid sequence comprising the VL region of an antibody and / or an amino acid sequence comprising its VH region (e.g., a light chain and / or a heavy chain of an antibody). The nucleic acid may encode one or more amino acid sequences comprising the VL region of an antibody and / or an amino acid sequence comprising its VH region (e.g., a light chain and / or a heavy chain of an antibody). In some embodiments, the nucleic acid, such as a polynucleotide, encodes one or more VH regions and / or one or more VL regions of an antibody in any order or orientation. In some embodiments, the nucleic acid, such as a polynucleotide, encodes the VH region and the VL region, and the coding sequence of the VH region is upstream of the coding sequence of the VL region. In some embodiments, the nucleic acid, such as a polynucleotide, encodes the VH region and the VL region, and the coding sequence of the VL region is upstream of the coding sequence of the VH region.

[0186] In some embodiments, one or more vectors (e.g., expression vectors) comprising such nucleic acids are provided. In another embodiment, a host cell comprising such nucleic acids is provided. In some embodiments, the host cell comprises (e.g., has been transformed as follows): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising an antibody VL region and an amino acid sequence comprising an antibody VH region, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising an antibody VL region and a second vector comprising a nucleic acid encoding an amino acid sequence comprising an antibody VH region. In some embodiments, the host cell comprises one or more vectors (e.g., transformed with the one or more vectors) comprising one or more nucleic acids encoding one or more amino acid sequences comprising one or more antibodies and / or portions thereof, such as antigen-binding fragments thereof. In some embodiments, one or more such host cells are provided. In some embodiments, a composition containing one or more such host cells is provided. In some embodiments, the one or more host cells can express different antibodies or the same antibody. In some embodiments, each of the host cells can express more than one antibody.

[0187] Also provided are methods for producing anti-PD-L1 antibodies (including antigen-binding fragments). For recombinant production of anti-PD-L1 antibodies, for example, nucleic acid sequences or polynucleotides encoding the antibodies as described above can be isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acid sequences can be easily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can specifically bind to genes encoding the heavy and light chains of the antibodies). In some embodiments, a method for producing an anti-PD-L1 antibody is provided, wherein the method comprises culturing a host cell comprising a nucleic acid sequence encoding the antibody as provided above under conditions suitable for expression of the antibody and optionally recovering the antibody from the host cell (or host cell culture medium).

[0188] In addition to prokaryotes, eukaryotic microorganisms, such as filamentous fungi or yeast, are suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been modified to mimic or approximate the glycosylation pathways in human cells, thereby producing antibodies with partially or fully human glycosylation patterns. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004) and Li et al., Nat. Biotech. 24: 210-215 (2006).

[0189] Exemplary eukaryotic cells that can be used to express polypeptides include, but are not limited to, COS cells, including COS 7 cells; 293 cells, including 293-6E cells; CHO cells, including CHO-S, DG44.Lec13 CHO cells, and FUT8 CHO cells; cells; and NSO cells. In some embodiments, antibody heavy and / or light chains (e.g., VH and / or VL regions) can be expressed in yeast. See, for example, U.S. Publication No. US2006 / 0270045 A1. In some embodiments, a particular eukaryotic host cell is selected based on its ability to perform the desired post-translational modification of the heavy and / or light chains (e.g., VH and / or VL regions). For example, in some embodiments, CHO cells produce polypeptides that are more sialylated than the same polypeptides produced in 293 cells.

[0190] In some embodiments, the antibodies or antigen-binding fragments provided herein are produced in a cell-free system. Exemplary cell-free systems are described, for example, in Sitaraman et al., Methods Mol. Biol. 498: 229-44 (2009); Spirin, Trends Biotechnol. 22: 538-45 (2004); Endo et al., Biotechnol. Adv. 21: 695-713 (2003).

[0191] The provided embodiments further include vectors and host cells and other expression systems for expressing and producing antibodies and other antigen-binding proteins, including eukaryotic and prokaryotic host cells, including bacteria, filamentous fungi and yeast, and mammalian cells, such as human cells, and cell-free expression systems.

[0192] 2. Exemplary Characteristics of Anti-PD-L1 Antibodies

[0193] In some aspects, antibodies or antigen-binding fragments thereof are provided that have one or more specific functional characteristics, such as binding properties, including binding to a specific epitope.

[0194] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to a PD-L1 protein. In some embodiments provided herein, the PD-L1 protein refers to human PD-L1, non-human primate (e.g., cynomolgus macaque) PD-L1 protein, or mouse PD-L1 protein. In some embodiments provided herein, the PD-L1 protein refers to human PD-L1 or non-human primate (e.g., cynomolgus macaque) PD-L1 protein. In some embodiments of any of the embodiments herein, the PD-L1 protein refers to human PD-L1 protein. Observing that an antibody or other binding molecule binds to a PD-L1 protein or specifically binds to a PD-L1 protein does not necessarily mean that it binds to PD-L1 proteins of every species. For example, in some embodiments, a characteristic of binding to a PD-L1 protein, such as the ability to specifically bind to it and / or bind to a particular degree with a particular affinity, in some embodiments refers to the ability relative to a human PD-L1 protein, and an antibody may not have this characteristic relative to a PD-L1 protein of another species (e.g., mouse).

[0195] In some embodiments, the antibody or antigen-binding fragment binds to a mammalian PD-L1 protein, including binding to naturally occurring PD-L1 variants, such as certain splice variants or allelic variants.

[0196] In some embodiments, the antibody specifically binds to the extracellular domain of a human PD-L1 protein, such as an epitope or region of a human PD-L1 protein, such as the extracellular domain of human PD-L1 comprising the amino acid sequence of SEQ ID NO: 327 (amino acid residues 19-239 of UniProt Accession No. Q9NZQ7), or an allelic variant or splice variant thereof. In some embodiments, the antibody binds to a cynomolgus macaque PD-L1 protein, such as the extracellular domain of the cynomolgus macaque PD-L1 set forth in SEQ ID NO: 328 (amino acid residues 19-239 of UniProt Accession No. G7PSE7). In some embodiments, the antibody binds to human PD-L1, but at a lower level or with a lower degree or affinity than a cynomolgus macaque PD-L1 protein, such as the extracellular domain of the cynomolgus macaque PD-L1 set forth in SEQ ID NO: 328 (amino acid residues 19-239 of UniProt Accession No. G7PSE7). In some embodiments, the antibody does not bind to or binds to a mouse PD-L1 protein at a lower level or with a lower degree or affinity, such as the mouse PD-L1 extracellular domain shown in SEQ ID NO: 329 (amino acid residues 19-238 of UniProt accession number Q9EP73). In some embodiments, the antibody binds to the mouse PD-L1 protein with a lower affinity than it binds to the human PD-L1 protein and / or the cynomolgus monkey PD-L1 protein. In some embodiments, the antibody binds to the mouse PD-L1 protein and / or the cynomolgus monkey PD-L1 protein with a lower affinity than it binds to the human PD-L1 protein. In some embodiments, the antibody binds to the cynomolgus monkey PD-L1 protein with a similar binding affinity as compared to its binding to the human PD-L1 protein.

[0197] In one embodiment, the extent of binding of an anti-PD-L1 antibody to an unrelated, non-PD-L1 protein, such as a non-human PD-L1 protein or other non-PD-L1 protein, is less than or about 10% of the binding of the antibody to the human PD-L1 protein, for example, as measured by radioimmunoassay (RIA), ELISA, or surface plasmon resonance (SPR). In some embodiments, the antibodies provided include antibodies that bind to a mouse PD-L1 protein that is less than, at or about 10% of the binding of the antibody to the human PD-L1 protein. In some embodiments, the antibodies provided include antibodies that bind to a cynomolgus monkey PD-L1 protein that is less than, at or about 50%, 40%, 30%, 20%, or 10% of the binding of the antibody to the human PD-L1 protein.

[0198] In some embodiments, provided antibodies are capable of binding to a PD-L1 protein, such as a human PD-L1 protein, with at least a certain affinity, as measured by any of a variety of known methods. In some embodiments, affinity is determined by the equilibrium dissociation constant (K D ) represents; in some embodiments, affinity is represented by EC 50 express.

[0199] A variety of assays can be used to assess binding affinity and / or determine whether a binding molecule (e.g., an antibody or fragment thereof) specifically binds to a particular ligand (e.g., an antigen, such as a PD-L1 protein). It is within the capabilities of those skilled in the art to determine the binding affinity of a binding molecule, such as an antibody, to an antigen (e.g., PD-L1, such as human PD-L1 or cynomolgus monkey PD-L1 or mouse PD-L1), such as by using any of a variety of binding assays well known in the art (see, e.g., Scatchard et al., Ann. NY Acad. Sci. 51:660, 1949; Wilson, Science 295:2103, 2002; Wolff et al., Cancer Res. 53:2560, 1993; and U.S. Pat. Nos. 5,283,173, 5,468,614, or equivalents).

[0200] For example, in some embodiments, The instrument can be used to determine the binding kinetics and constants of a complex between two proteins (e.g., an antibody, such as an anti-PD-L1 antibody or fragment thereof, and an antigen, such as a PD-L1 protein) using surface plasmon resonance (SPR) analysis. SPR measures the change in the concentration of molecules on the sensor surface as the molecules bind to or dissociate from the surface. The change in the SPR signal is proportional to the change in mass concentration close to the surface, allowing the binding kinetics between the two molecules to be measured. The dissociation constant of the complex can be determined by monitoring the change in refractive index over time as the buffer passes over the chip.

[0201] Other suitable assays for measuring the binding of one protein to another include, for example, immunoassays such as enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), or by monitoring changes in the spectral or optical properties of the protein by fluorescence, UV absorption, circular dichroism, or nuclear magnetic resonance (NMR). Other exemplary assays include, but are not limited to, Western blotting, analytical ultracentrifugation, spectroscopy, flow cytometry, and other methods for detecting protein binding.

[0202] In some embodiments, provided antibodies, fragments, or conjugates bind to a PD-L1 protein, such as a human PD-L1 protein, with similar or greater affinity than a reference antibody (e.g., avelumab). In some embodiments, binding affinity is measured as the half maximal effective concentration (EC 50 ) or dissociation constant (K D ECs that bind to free or immobilized PD-L1, such as PD-L1 expressed on the cell surface or the extracellular domain of PD-L1, can be determined. 50 or K D In some embodiments, provided antibodies, fragments, or ECs of conjugates to PD-L1 proteins, such as human PD-L1 proteins, are 50 or K D The value is similar to or lower than (e.g., corresponding to a higher binding affinity) a reference antibody (e.g., avelumab).

[0203] In some embodiments, binding affinity is measured as the half maximal effective concentration (EC 50 In some embodiments, the EC of an antibody, fragment, or conjugate that binds to a PD-L1 protein (such as a human PD-L1 protein) 50 Similar to or less than the EC of a reference antibody, fragment or conjugate (e.g., avelumab or its corresponding fragment or conjugate) 50 In some embodiments, the EC of the antibody, fragment, or conjugate 50 A decrease of about 0.5 times, about 1 times, about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times or more.

[0204] In some embodiments, binding affinity is measured as the K of the antibody, fragment, or conjugate. D In some embodiments, the K of an antibody, fragment, or conjugate for binding to a PD-L1 protein (such as a human PD-L1 protein) is D Similar to or less than the K of a reference antibody, fragment or conjugate (e.g., avelumab or its corresponding fragment or conjugate) D In some embodiments, the K of the antibody, fragment, or conjugate is DA decrease of about 0.5 times, about 1 times, about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, about 10 times, about 15 times, about 20 times, about 25 times, about 30 times, about 35 times, about 40 times, about 45 times, about 50 times or more.

[0205] In some embodiments, the binding affinities of binding molecules, such as anti-PD-L1 antibodies, to different antigens, such as PD-L1 proteins from different species, can be compared to determine species cross-reactivity. For example, species cross-reactivity can be classified as high cross-reactivity or low cross-reactivity. In some embodiments, the equilibrium dissociation constants K for different antigens, such as PD-L1 proteins from different species, such as from humans, cynomolgus monkeys, or mice, can be compared. D or EC 50 To determine species cross-reactivity. In some embodiments, the species cross-reactivity of the anti-PD-L1 antibody can be high, for example, the anti-PD-L1 antibody binds to human PD-L1 and species variant PD-L1 to a similar extent, for example, the EC of human PD-L1 is 50 or K D EC with species variant PD-L1 50 or K D The ratio of is or is about 1. In some embodiments, the species cross-reactivity of the anti-PD-L1 antibody may be low, for example, the anti-PD-L1 antibody has a high affinity for human PD-L1 but a low affinity for species variant PD-L1, or vice versa. The degree of species cross-reactivity can also be compared with the species cross-reactivity of known antibodies.

[0206] In some embodiments, provided antibodies or antigen-binding fragments thereof bind to human PD-L1 protein and non-human PD-L1 protein or other non-PD-L1 protein to similar extents. For example, in some embodiments, provided antibodies or antigen-binding fragments thereof bind to a certain EC 50 or K D ECs that bind to human PD-L1 proteins, such as those comprising the amino acid sequence of SEQ ID NO: 327 (amino acid residues 19-239 of UniProt Accession No. Q9NZQ7) or an allelic variant or splice variant thereof, and non-human PD-L1, such as cynomolgus macaque PD-L1, such as the cynomolgus macaque PD-L1 protein set forth in SEQ ID NO: 328 (amino acid residues 19-239 of UniProt Accession No. G7PSE7). 50 or K DIn other embodiments, the antibodies or antigen-binding fragments thereof provided herein bind to a human PD-L1 protein, such as an EC of a human PD-L1 protein comprising the amino acid sequence of SEQ ID NO: 327 (amino acid residues 19-239 of UniProt accession number Q9NZQ7) or an allelic variant or splice variant thereof. 50 or K D Lower than the EC of non-human PD-L1, such as mouse PD-L1, such as the mouse PD-L1 protein shown in SEQ ID NO: 329 (amino acid residues 19-238 of UniProt accession number Q9EP73). 50 or K D .

[0207] In some embodiments, the antibody (e.g., antigen binding fragment) exhibits a binding preference for cells expressing PD-L1, such as specific cells expressing PD-L1 and known not to express PD-L1, as compared to PD-L1 negative cells. In some embodiments, the binding preference is observed when the degree of binding to cells expressing PD-L1 is significantly greater than that to cells not expressing it is measured. In some embodiments, the binding preference is measured, for example, by the mean fluorescence intensity and / or dissociation constant or EC in an analysis based on flow cytometry. 50 The fold change in the extent of binding detected in cells expressing PD-L1 compared to cells not expressing PD-L1 is measured to be at least or about 1.5, 2, 3, 4, 5, 6, or more, and / or is about the same as, about the same as, at least the same as, or at least about the same as, or greater than the fold change observed for the corresponding form of the reference antibody.

[0208] The anti-PD-L1 antibodies (e.g., antigen binding fragments) provided herein can be identified, screened, or characterized for their physical / chemical properties and / or biological activity via various known assays. In one aspect, the antigen binding activity of the antibody is tested, for example, by known methods such as ELISA, Western blot, and / or flow cytometric analysis, including cell-based binding assays, such as assessing the binding of the antibody (e.g., conjugated to a fluorescent marker or spiked) to cells expressing the target antigen (e.g., PD-L1), in some cases compared to the results obtained using cells that do not express the target antigen (e.g., PD-L1). Binding affinity can be K D or EC 50 Measurement. In some examples, binding affinity, binding kinetics, and / or binding constants can be measured using assays to determine molecular interactions, such as surface plasmon resonance analysis.

[0209] In some aspects, the anti-PD-L1 antibodies (e.g., antigen-binding fragments) provided herein can be conjugated to one or more agents, such as one or more drugs, small molecules, dyes (e.g., phthalocyanine dyes such as IR700) and / or therapeutic agents, e.g., as described in Section II.

[0210] In some aspects, after conjugation to one or more agents, the conjugate comprising the provided antibody or antigen-binding fragment retains or substantially retains its physical / chemical properties and / or biological activity such that the physical / chemical properties and / or biological activity of the conjugate are equal to or similar to the physical / chemical properties and / or biological activity of the unconjugated (naked) antibody or antigen-binding fragment, or their physical / chemical properties and / or biological activity are reduced by no more than 50%, no more than 40%, no more than 30%, no more than 20%, no more than 10%, no more than 5%, no more than 4%, no more than 3%, no more than 2%, or no more than 1%, or at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% compared to the physical / chemical properties and / or biological activity of the unconjugated (naked) antibody.

[0211] In some aspects, after conjugation with one or more agents, such as phthalocyanine dyes, such as IR700, the conjugate comprising the provided antibody or antigen-binding fragment retains or substantially retains binding affinity for the target antigen PD-L1, such that the binding affinity of the conjugate is equal to or similar to the binding affinity of the unconjugated (naked) antibody or antigen-binding fragment, or the binding affinity is reduced by no more than 50%, no more than 40%, no more than 30%, no more than 20%, no more than 10%, no more than 5%, no more than 4%, no more than 3%, no more than 2% or no more than 1%, or exhibits at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% of the binding affinity of the unconjugated (naked) antibody. In some embodiments, the conjugate exhibits similar binding to the PD-L1 protein compared to the unconjugated antibody.

[0212] Competition assays can be used to identify antibodies that compete with any of the antibodies (e.g., antigen-binding fragments) described herein or with other known anti-PD-L1 antibodies. Assays for locating the epitope to which the antibodies and other known anti-PD-L1 antibodies bind can also be used and are known.

[0213] B. Multispecific Antibodies

[0214] In certain embodiments, PD-L1 binding molecules, such as antibodies or polypeptides, such as chimeric receptors containing them, are multispecific. Among the multispecific binding molecules are multispecific antibodies, including, for example, bispecific or trispecific antibodies. A multispecific binding partner (e.g., an antibody) has binding specificities for at least two different sites that may be on the same or different antigens. In certain embodiments, one of the binding specificities is for PD-L1 and the other is for another antigen. In certain embodiments, a bispecific antibody comprises an additional binding domain that is specific for a second or additional antigen. In some embodiments, the additional binding molecule binds to and / or recognizes a third or more antigens. In certain embodiments, a bispecific antibody can bind to two different epitopes of PD-L1. Bispecific antibodies can also be used to localize fluorescent agents, phototoxic agents, and / or cytotoxic agents to cells expressing PD-L1. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments. Among the multispecific antibodies are multispecific single-chain antibodies, such as diabodies, triabodies, and tetrabodies, tandem di-scFvs, and tandem tri-scFvs.

[0215] In some embodiments, the second or additional antigens used in the multi-targeting strategy include at least one of the antigens being a universal tumor antigen or a family member thereof. In some embodiments, the second or additional antigen is an antigen expressed on a tumor. In some embodiments, the PD-L1 binding molecules provided herein target an antigen on the same tumor type as the second or additional antigen. In some embodiments, the second or additional antigen may also be a universal tumor antigen or a tumor antigen specific to a tumor type.

[0216] Exemplary second or additional antigens include CD4, CD5, CD8, CD14, CD15, CD19, CD20, CD21, CD22, CD23, CD25, CD33, CD37, CD38, CD40, CD40L, CD46, CD47, CD52, CD54, CD74, CD80, CD126, CD138, CTLA-4, B7, MUC-1, Ia, HM1.24, HLA-DR, tenascin, angiogenic factor, VEGF, PIGF, SIRPα, ED-B fibronectin, oncogenes, oncogene products, CD66a-d, necrosis antigens, Ii, IL-2, T101, TAC, IL-6, ROR1, TRAIL-R1 (DR4), TRAIL-R2 (DR5), tEGFR, Her2, L1-CAM, mesothelin, CEA, hepatitis B surface antigen, antifolate receptor, CD24, CD30, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, ErbB3, ErbB4, erbB dimer, EGFR vIII, FBP, FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, G protein-coupled receptor class C group 5 member D (GPRC5D), HMW-MAA, IL-22R-α, IL-13R-α2, kdr, kappa light chain, Lewis Y, L1-cell adhesion molecule (L1-CAM), melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, preferentially expressed antigen in melanoma (PRAME), survivin, EGP2, EGP40, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, CD171, G250 / CAIX, HLA-AI MAGE A1, HLA-A2NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7 / 8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, fetal AchR, NKG2D ligand, dual antigens, antigens associated with universal signatures, cancer-testis antigen, MUC1, MUC16, NY-ESO-1, MART-1, gp100, carcinoembryonic antigen, VEGF-R2, carcinoembryonic antigen (CEA), prostate-specific antigen, PSMA, Her2 / neu, estrogen receptor, progesterone receptor, ephrin B2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms' tumor 1 (WT-1), cyclin, cyclin A2, CCL-1, hTERT, MDM2, CYP1B, WT1, livin, AFP, p53, cyclin (D1), CS-1, BAFF-R, TACI, CD56, TIM-3, CD 123, L1-cell adhesion molecule, MAGE-A1, MAGE A3, cyclin, such as cyclin A1 (CCNA1) and / or pathogen-specific antigens, biotinylated molecules, molecules expressed by HIV, HCV, HBV and / or other pathogens and / or in some aspects, neo-epitopes or neo-antigens thereof. In some embodiments, the antigen is associated with a universal tag or is a universal tag.

[0217] In some aspects, antigens (e.g., second or additional antigens, such as disease-specific antigens and / or associated antigens) are expressed on multiple myeloma, such as G protein-coupled receptor class C 5 member D (GPRC5D), CD38 (cyclic ADP ribose hydroxylase), CD138 (syndecan-1, syndecan, SYN-1), CS-1 (CS1, CD2 subset 1, CRACC, SLAMF7, CD319 and 19A24), BAFF-R, TACI and / or FcRH5. Other exemplary multiple myeloma antigens include CD56, TIM-3, CD33, CD123, CD44, CD20, CD40, CD74, CD200, EGFR, β2-microglobulin, HM1.24, IGF-1R, IL-6R, TRAIL-R1 and activin receptor type IIA (ActRIIA). See Benson and Byrd, J. Clin. Oncol. (2012) 30(16):2013-15; Tao and Anderson, Bone Marrow Research (2011):924058; Chu et al., Leukemia (2013) 28(4):917-27; Garfall et al., Discov Med. (2014) 17(91):37-46. In some embodiments, the antigen includes an antigen present on lymphoid tumors, myeloma, AIDS-related lymphoma and / or post-transplant lymphoproliferation, such as CD38. Antibodies or antigen-binding fragments directed against such antigens are known and include, for example, those described in U.S. Patent Nos. 8,153,765; 8,603,477; 8,008,450; U.S. Publication Nos. US20120189622 or US20100260748; and / or International PCT Publication Nos. WO2006099875; WO2009080829; WO2012092612; or WO2014210064. In some embodiments, such antibodies or antigen-binding fragments thereof (e.g., scFv) are contained in multispecific antibodies, multispecific chimeric receptors, and / or multispecific cells.

[0218] C. Variants

[0219] In certain embodiments, an antibody (e.g., an antigen binding fragment) includes one or more amino acid changes, such as substitutions, deletions, insertions, and / or mutations, compared to the sequences of the antibodies described herein. Exemplary variants include variants designed to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of an antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and / or insertions and / or substitutions of residues within the amino acid sequence of the antibody. Any combination of deletions, insertions, and substitutions can be performed to obtain the final construct, subject to the proviso that the final construct has the desired characteristics, such as antigen binding.

[0220] In certain embodiments, the antibody (e.g., antigen-binding fragment) comprises one or more amino acid substitutions, e.g., compared to the antibody sequences described herein and / or compared to the sequence of a natural repertoire (e.g., a human repertoire). Relevant sites for substitutional mutagenesis include CDRs and FRs. Amino acid substitutions can be introduced into the antibody of interest and the products screened for the desired activity, e.g., retained / improved antigen binding, reduced immunogenicity, improved half-life, and / or improved effector function, such as the ability to promote antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC).

[0221] In some embodiments, one or more residues within the CDR of a parent antibody (e.g., a humanized antibody or a human antibody) are substituted. In some embodiments, substitutions are made to restore the sequence or position in the sequence to a germline sequence, such as an antibody sequence found in a germline (e.g., a human germline), e.g., to reduce the potential for immunogenicity, e.g., when administered to a human subject.

[0222] In certain embodiments, substitutions, insertions or deletions may occur within one or more CDRs, as long as such changes do not substantially reduce the ability of the antibody to bind to the antigen. For example, conservative changes that do not substantially reduce binding affinity (e.g., conservative substitutions as provided herein) may be made in the CDRs. Such changes may be made, for example, outside of the residues that contact the antigen in the CDRs. In certain embodiments of the variant VH and VL region sequences provided above, each CDR is unchanged or contains no more than one, two or three amino acid substitutions.

[0223] Amino acid sequence insertions include amino-terminal and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing one hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertional variants of the antibody molecule include fusions of the N- or C-terminus of the antibody to enzymes or polypeptides that increase the serum half-life of the antibody.

[0224] D. Modification

[0225] In certain embodiments, the antibody is altered to increase or decrease the extent of glycosylation of the antibody, for example, by changing the amino acid sequence and / or by modifying the oligosaccharides attached to the glycosylation site (e.g., using certain cell lines) to remove or insert one or more glycosylation sites. In some embodiments, N-linked glycosylation is removed or inserted, which is a glycosylation site that occurs at an asparagine residue in the consensus sequence -Asn-Xaa-Ser / Thr.

[0226] Exemplary modifications, variants, and cell lines are described in, for example, patent publications Nos. US 2003 / 0157108, US 2004 / 0093621, US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2003 / 085119. No. 2005 / 035586; No. WO 2005 / 035778; No. WO 2005 / 053742; No. WO 2002 / 031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US 2003 / 0157108A1, Presta, L; and WO 2004 / 056312 A1, Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al. Biotechnol. Bioeng., 94(4):680-688 (2006); and WO 2003 / 085107); WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.); WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0227] A modified antibody is an antibody having one or more amino acid modifications in the Fc region, such as an antibody having a human Fc region sequence or other portion of a constant region (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) comprising an amino acid modification (e.g., substitution) at one or more amino acid positions. Such modifications can be made, for example, to improve half-life, alter binding to one or more types of Fc receptors, and / or alter effector function.

[0228] In some of any of the provided embodiments, the anti-PD-L1 antibody comprises a functional Fc region. In some of any of the provided embodiments, the anti-PD-L1 antibody comprises a full-length Fc region. In some embodiments, the anti-PD-L1 antibody comprises an IgG1 Fc region. In some embodiments, the anti-PD-L1 antibody comprises an IgG2 Fc region. In some embodiments, the IgG2 Fc region is an IgG2a Fc region. In some embodiments, the IgG2 Fc region is an IgG2a / b Fc region. In some embodiments, the IgG2 Fc region is an IgG2a Fc region. In some of any of the provided embodiments, the anti-PD-L1 antibody comprises an IgG3 Fc region. In some of any of the provided embodiments, the anti-PD-L1 antibody comprises an IgG4 Fc region. In some embodiments, the Fc region is modified to modulate the effector function of the antibody portion of the conjugate. Any of such modifications, such as those described in Wang et al., (2018) Protein Cell. 9(1):63-73, are contemplated for use in the antibodies, antibody fragments, and conjugates described herein.

[0229] In some of any of the provided embodiments, the anti-PD-L1 antibody does not comprise a functional Fc region. In some of such examples, the anti-PD-L1 antibody does not contain an Fc region or comprises an Fc region that has been modified such that the Fc region does not bind to an Fc receptor and / or does not elicit substantial Fc effector functions (e.g., ADCC, ADCP, and / or CDC). In some of such embodiments, the anti-PD-L1 antibody comprises an Fc receptor comprising an amino acid substitution that eliminates the glycosylation site at a position corresponding to position 297 of the heavy chain based on EU numbering as described in Edelman et al. (1969) Proc Natl Acad Sci US A. 63(1):78-85. For example, the anti-PD-L1 antibody may comprise an Fc receptor containing an asparagine to glutamine substitution (N297Q) at position 297 or corresponding to position 297, by EU numbering relative to the antibody sequence, an asparagine to alanine substitution (N297G) at position 297 or corresponding to position 297, by EU numbering relative to the antibody sequence, or an asparagine to glycine substitution (N297G) at position 297 or corresponding to position 297, by EU numbering relative to the antibody sequence.

[0230] In some of any of the provided embodiments, the anti-PD-L1 antibody comprises an Fc region that exhibits enhanced Fc-mediated effector function (such as ADCC, ADCP, and / or CDC activity) and / or exhibits preferential binding to Fcγ receptors. In some embodiments, the anti-PD-L1 antibody exhibits enhanced function due to increased Fc receptor engagement. In some embodiments, the Fc region contains one or more of the following mutations: a glycine to alanine substitution at position 236 (G236A), a serine to aspartic acid substitution at position 239 (S239D), an alanine to leucine substitution at position 330 (A330L), an isoleucine to glutamic acid substitution at position 332 (I332E), a glutamic acid to alanine substitution at position 333 (E333A), a lysine to alanine substitution at position 334 (K334A), an arginine to alanine substitution at position 255 (S255A), a threonine to alanine substitution at position 256 (T336A), relative to EU numbering of the antibody heavy chain. 256A), a serine to alanine substitution at position 267 (S267A), a serine to alanine substitution at position 298 (S298A), an asparagine to serine substitution at position 325 (N325S), a leucine to phenylalanine (L328F) at position 328, an alanine to leucine substitution at position 330 (A330L), an isoleucine to glutamic acid substitution at position 333 (E333A), a glutamic acid to alanine substitution at position 333 (E333A), a lysine to alanine substitution at position 334 (K334), and / or an alanine to glutamine substitution at position 378 (A378Q). In some embodiments, the Fc region comprises a glycine to alanine substitution at position 236, a serine to aspartic acid substitution at position 239, and an isoleucine to glutamic acid substitution at position 332 (G236A / S239D / I332E), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises a glycine to alanine substitution at position 236, an alanine to leucine substitution at position 330, and an isoleucine to glutamic acid substitution at position 332 (G236A / A330L / I332E), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a glycine to alanine substitution at position 236, a serine to aspartic acid substitution at position 239, an alanine to leucine substitution at position 330, and an isoleucine to glutamic acid substitution at position 332 (G236A / S239D / A330L / I332E), relative to EU numbering of the antibody heavy chain.In some embodiments, the Fc region comprises a substitution of serine at position 239 to aspartic acid and isoleucine at position 332 to glutamic acid (S239D / I332E), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises a substitution of serine at position 298 to alanine and lysine at position 334 to alanine (S298A / K334), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises a substitution of glutamic acid at position 333 to alanine and lysine at position 334 to alanine (E333A / K334), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises a substitution of serine at position 298 to alanine, glutamic acid at position 333 to alanine, and lysine at position 334 to alanine (S298A / E333A / K334), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a substitution of arginine at position 255 to alanine and a substitution of serine at position 267 to alanine (R255A / S267A) relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a substitution of threonine at position 256 to alanine (T256A) relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a substitution of lysine at position 334 to alanine and a substitution of alanine at position 378 to glutamine (K334 / A378) relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a substitution of serine at position 239 to aspartic acid, a substitution of alanine at position 330 to leucine, and a substitution of isoleucine at position 332 to glutamic acid (S239D / A330L / I332E) relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises a glycine to alanine substitution at position 236, a serine to aspartic acid substitution at position 239, and an isoleucine to glutamic acid substitution at position 332 (G236A / S239D / I332E), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region comprises an arginine to serine substitution at position 325 and a leucine to phenylalanine substitution at position 328 (N325S / L328F), relative to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a phenylalanine to leucine substitution at position 243, an arginine to proline substitution at position 292, a tyrosine to leucine substitution at position 300, a valine to isoleucine substitution at position 305, and a proline to leucine substitution at position 396 (F243L / R292P / Y300L / V305I / P396L), relative to EU numbering of the antibody heavy chain.In some embodiments, the Fc region comprises, relative to the EU numbering of an antibody heavy chain, a leucine to valine substitution at position 235, a phenylalanine to leucine substitution at position 243, an arginine to proline substitution at position 292, a tryptophan to leucine substitution at position 300, and a proline to leucine substitution at position 396 (L235V / F243L / R292P / Y300L / P396L). In some embodiments, the Fc region comprises, according to EU numbering of one of the antibody heavy chains, a leucine to tyrosine substitution at position 234, a leucine to glutamine substitution at position 235, a glycine to tryptophan substitution at position 236, a serine to methionine substitution at position 239, a histidine to aspartic acid substitution at position 268, an aspartic acid to glutamic acid substitution at position 270, and a serine to alanine substitution at position 298. The heavy chain comprises a amino acid sequence of the ...

[0231] Variants are also cysteine ​​engineered antibodies, such as "thioMAbs" and other cysteine ​​engineered variants, in which one or more residues of an antibody are substituted with cysteine ​​residues to create reactive thiol groups at accessible sites, for example, for conjugation of agents and linker-agents to create immunoconjugates. Cysteine ​​engineered antibodies are described, for example, in U.S. Patent Nos. 7,855,275 and 7,521,541.

[0232] In some embodiments, antibodies (e.g., antigen-binding fragments) are modified to contain additional non-proteinaceous moieties, including water-soluble polymers. Exemplary polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, polydextrose, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers), and polydextrose or poly(n-vinyl pyrrolidone) polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer may have any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if more than one polymer is attached, the polymers may be the same or different molecules. Generally, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular property or function of the antibody to be improved, whether the antibody derivative will be used for therapy in a given condition, and the like.

[0233] In some embodiments, the polynucleotide is optimized or contains certain features designed for optimization (such as for codon usage) and / or modification, for example, to increase expression (such as surface expression) of the encoded antibody or fragment or to make its expression more consistent between cell product batches. In some embodiments, the polynucleotide encoding the anti-PD-L1 antibody is modified to remove cryptic or hidden splice sites.

[0234] II. Conjugates

[0235] In some embodiments, an anti-PD-L1 antibody or antibody fragment (e.g., an antigen-binding fragment) is or is part of an immunoconjugate in which the antibody is conjugated to one or more heterologous molecules or moieties, such as, but not limited to, a phototoxic agent, a cytotoxic agent, or an imaging agent. In specific embodiments, the antibody is conjugated to a phototoxic agent, such as a phthalocyanine dye. In some embodiments, the antibody is conjugated to a phototoxic agent (such as a phthalocyanine dye) and a second phototoxic agent, cytotoxic agent, or imaging agent.

[0236] A. Cytotoxic Agents

[0237] Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, and Lu radioisotopes); chemotherapeutic agents (e.g., methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other intercalating agents); growth inhibitory agents; enzymes and fragments thereof, such as ribolytic enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins. In some embodiments, the antibody is conjugated to one or more cytotoxic agents, such as chemotherapeutic agents or drugs, growth inhibitory agents, toxins (e.g., protein toxins, enzymatically active toxins of bacterial, fungal, plant, or animal origin), or radioactive isotopes.

[0238] Among the immunoconjugates are antibody-drug conjugates (ADCs) in which an antibody is conjugated to one or more drugs, including but not limited to a maytansinoid; an auristatin, such as the monomethyl auristatin drug moieties DE and DF (MMAE and MMAF; dolastatin; calicheamicin or a derivative thereof; an anthracycline, such as daunomycin or doxorubicin; methotrexate; vindesine; a taxane, such as docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; trichothecene; and CC1065.

[0239] Also included among the immunoconjugates are those in which the antibody is conjugated to an enzymatically active toxin or fragment thereof, including but not limited to diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis toxin, scutellaria baicalensis ... officinalis inhibitors, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and tricothecenes.

[0240] Non-limiting, exemplary cytotoxic agents include aplidin, azaribine, anastrozole, azacytidine, bleomycin, bortezomib, bryostatin-1, busulfan, calicheamycin, camptothecin, 10-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin, and salidomide. atin), irinotecan (CPT-11), SN-38, carboplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dactinomycin, daunorubicin glucuronide, daunomycin, dexamethasone, diethylstilbestrol, doxorubicin, doxorubicin glucuronide, epirubicin glucuronide, ethinyl estradiol, estramustine, etoposide, etoposide glucuronide, etoposide phosphate, floxuridine (FUdR), 3',5'-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone acetate, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, leucovorin, lomustine, nitrogen mustard, medroxyprogesterone acetate, megestrol acetate, melphalan, mercapto Purine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, phenyl butyrate, prednisone, procarbazine, paclitaxel, pentostatin, PSI-341, semustine, streptozotocin, streptozocin), tamoxifen, taxanes, paclitaxel, testosterone propionate, thalidomide, thioguanine, thiotepa, teniposide, topotecan, uracil mustard, velcade, vinblastine, vinorelbine, vincristine, ricin, abrin, ribomiclease, onconase, rapLR1, DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin.

[0241] Also included among immunoconjugates are those in which an antibody (eg, an antigen-binding fragment) is conjugated to a radioactive atom to form a radioconjugate. Exemplary radioisotopes include At 211 , I 131 , I 125 、Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 、P 32 , Pb 212 And radioactive isotopes of Lu.

[0242] B. Phototoxic agents

[0243] In certain aspects, an anti-PD-L1 antibody or antibody fragment (e.g., an antigen-binding fragment) is conjugated to a toxic agent. In some aspects, the phototoxic agent is a phthalocyanine dye. Phthalocyanines are a group of photosensitizer compounds having a phthalocyanine ring system. Phthalocyanines are azaporphyrins (i.e., C32H18N8) containing four benzindole groups connected by nitrogen bridges in a 16-membered ring of alternating carbon and nitrogen atoms, which form stable chelates with metal or metalloid cations. In these compounds, the ring center is occupied by a metal ion (diamagnetic or paramagnetic), which, depending on the ion, can carry zero, one, or two ligands. In addition, the ring periphery can be unsubstituted or substituted.

[0244] In some embodiments, phthalocyanines strongly absorb red or near-IR radiation, with an absorption peak between approximately 600 nm and 810 nm, which in some cases allows light to penetrate deep into tissue. Phthalocyanines are generally photostable. This photostability is generally advantageous in pigments and dyes, and in a variety of other phthalocyanine applications.

[0245] In some embodiments, the phthalocyanine dye is water-soluble and contains a luminescent fluorophore moiety having at least one water-solubilizing moiety. In some embodiments, the water-solubilizing moiety contains silicon. In some embodiments, the phthalocyanine dye has a core atom such as Si, Ge, Sn, Al, or Zn. In some embodiments, the phthalocyanine dye contains a linker having a reactive group capable of forming a bond between the linker and another molecule, i.e., forming a conjugate. In some embodiments, the phthalocyanine dye can be tuned to fluoresce at a specific wavelength.

[0246] In some embodiments, the phthalocyanine dye contains a linker, ie, a linker-phthalocyanine dye moiety (LD).

[0247] In some embodiments, the phthalocyanine dye is a phthalocyanine dye with a silicon coordinating metal (Si-phthalocyanine dye). In some embodiments, the phthalocyanine dye comprises the following formula:

[0248] or a salt, stereoisomer or tautomer thereof, wherein:

[0249] L is the connector;

[0250] Q is a reactive group used to link the dye to the targeting molecule;

[0251] R 2 、R 3 、R 7 and R 8 are each independently selected from optionally substituted alkyl and optionally substituted aryl;

[0252] R 4、R 5 、R 6 、R 9 、R 10 and R 11 are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkanoyl, optionally substituted alkoxycarbonyl, optionally substituted alkylcarbamoyl and a chelating ligand, wherein R 4 、R 5 、R 6 、R 9 、R 10 and R 11 At least one of the compounds comprises a water-soluble group;

[0253] R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18 、R 19 、R 20 、R 21 、R 22 and R 23 are each independently selected from hydrogen, halogen, optionally substituted alkylthio, optionally substituted alkylamino, and optionally substituted alkoxy; and

[0254] X 2 and X 3 are each independently C1-C1 optionally interrupted by heteroatoms 10 Alkylene.

[0255] In some embodiments, the phthalocyanine dye comprises the formula:

[0256] Its salts, stereoisomers and tautomers, wherein:

[0257] X 1 and X 4 are each independently C1-C1 optionally interrupted by heteroatoms 10 alkylene;

[0258] R 2 、R 3 、R 7 and R 8 are each independently selected from optionally substituted alkyl and optionally substituted aryl;

[0259] R 4 、R 5 、R 6 、R 9 、R 10 and R 11are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkanoyl, optionally substituted alkoxycarbonyl, optionally substituted alkylcarbamoyl and a chelating ligand, wherein R 4 、R 5 、R 6 、R 9 、R 10 and R 11 At least one of comprises a water-soluble group; and

[0260] R 16 、R 17 、R 18 and R 19 are each independently selected from hydrogen, halogen, optionally substituted alkylthio, optionally substituted alkylamino, and optionally substituted alkoxy.

[0261] In some embodiments of the methods and uses provided herein, the Si-phthalocyanine dye is IRDye700DX (IR700). In some embodiments, the phthalocyanine dye containing a reactive group is IR700NHS ester, such as IRDye 700DXNHS ester (LiCor 929-70010, 929-70011). In some embodiments, the dye is a compound having the formula:

[0262] , or a salt, stereoisomer or tautomer thereof.

[0263] For purposes herein, the term "IR700," "IRDye 700," or "IRDye 700DX" includes the above formula when the dye is conjugated, for example, via a reactive group, such as to an antibody.

[0264] In some embodiments, the phthalocyanine dye contains a linker, i.e., a linker-phthalocyanine dye moiety (LD). In some embodiments, the linker contains a reactive group. In one aspect, the phthalocyanine dye compound has formula (X):

[0265] or a salt, stereoisomer or tautomer thereof, wherein:

[0266] M is a metal or metalloid;

[0267] X is

[0268] Y is

[0269] R 1 and R 2each independently is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl;

[0270] R 3 、R 4 or R 5 is selected from substituent (a) or substituent (b), wherein

[0271] (a)R 3 For hydrogen, -L 3 -H, -L 3 -A or -L 3 -Z;

[0272] R 4 -L 4 -H, -(NH) m -L 4 -A, -(NH) m -L 4 -Z, -(O) m -L 4 -A or -(O) m -L 4 -Z;

[0273] R 5 -L 5 -H or -L 5 -A; and

[0274] (b)R 3 -L 3 -H or -L 3 -A;

[0275] R 4 -L 4 -H, -(NH) m -L 4 -A or -(O) m -L 4 -A; where R 3 and R 4 Connected to form -L 4 -A substituted heterocyclyl; and

[0276] R 5 -L 5 -H or -L 5 -A;

[0277] The premise is that R 3 、R 4 and R 5 At least one of them is a group containing A;

[0278] A is a reactive group capable of forming a covalent bond with a thiol, hydroxyl, carboxyl or amino group of the second moiety, or a protected form thereof or a reactive form thereof;

[0279] R 6 and R 7 each independently is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl;

[0280] R 8 、R 9 or R 10 is selected from substituent (a) or substituent (b), wherein

[0281] (a)R 8 For hydrogen, -L 8 -H or -L 8 -Z;

[0282] R 9 -L 9 -H, -(NH) n -L 9 -Z or -(O) n -L 9 -Z;

[0283] R 10 -L 10 -Z; and

[0284] (b)R 8 and R 9 Connected to form -L 9 -Z substituted heterocyclic group and R 10 -L 10 -H or -L 10 -Z;

[0285] The premise is that R 8 、R 9 and R 10 At least one of them is a group containing Z;

[0286] Z is a water-soluble group optionally substituted with A or L'-A;

[0287] L 1 and L 2 each independently is optionally substituted alkylene, optionally substituted heteroalkylene, optionally substituted alkenylene, optionally substituted heteroalkenylene, optionally substituted cycloalkyl, or optionally substituted heterocyclyl;

[0288] L3 , L 4 , L 5 , L 8 , L 9 and L 10 each independently is optionally substituted alkylene, optionally substituted heteroalkylene, optionally substituted alkenylene, optionally substituted heteroalkenylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted aralkylene, optionally substituted heteroaralkylene, or optionally substituted heteroarylene, wherein a carbon atom of the alkylene, heteroalkylene, alkenylene, heteroalkenylene, cycloalkylene, heterocyclylene, arylene, aralkylene, heteroaralkylene, or optionally substituted heteroarylene is further optionally substituted with Z, and each nitrogen atom of the heteroalkylene or heteroalkenylene is optionally substituted with one or two L′-Z;

[0289] each L' is independently optionally substituted alkylene, optionally substituted heteroalkylene, optionally substituted alkenylene, optionally substituted heteroalkenylene, optionally substituted cycloalkylene, optionally substituted heterocycloene, optionally substituted arylene, optionally substituted aralkylene, optionally substituted heteroaralkylene, or optionally substituted heteroarylene;

[0290] a is 0 or 1;

[0291] b is 0 or 1;

[0292] c is 0 or 1;

[0293] d is 0 or 1;

[0294] m is 0 or 1;

[0295] n is 0 or 1;

[0296] The premise is that if b is 1, then a is 0;

[0297] If d is 1, then c is 0;

[0298] If m is 1, then b is 1; and

[0299] If n is 1, then c is 1.

[0300] In certain embodiments, M is Si, Ge, Sn, or Al. In certain embodiments, M is Si or Ge.

[0301] In certain embodiments, provided herein are conjugates comprising a compound of formula (X) selected from the group consisting of:

[0302]

[0303]

[0304]

[0305] or a salt, stereoisomer or tautomer thereof. In a specific embodiment, the compound of formula (X) is selected from the following group:

[0306] or a salt, stereoisomer or tautomer thereof.

[0307] In some embodiments, the phthalocyanine dye is selected from Table A.

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314] In a specific embodiment, the phthalocyanine dye containing a reactive group has a structure of formula (I):

[0315] or a salt, stereoisomer or tautomer thereof.

[0316] In a specific embodiment, the phthalocyanine dye containing a reactive group has a structure of formula (II):

[0317] or a salt, stereoisomer or tautomer thereof.

[0318] C. Other features

[0319] In some embodiments, provided herein are conjugates and conjugates for use in the methods herein, comprising a Si-phthalocyanine dye attached to an anti-PD-L1 antibody or fragment provided herein. In some embodiments, the conjugate is an anti-PD-L1 antibody-Si-phthalocyanine dye conjugate. In some embodiments, the conjugate is an anti-PD-L1 antibody-IR700 conjugate.

[0320] In some embodiments, the anti-PD-L1 antibody or anti-PD-L1 antibody conjugate provided contains a functional Fc region. In some embodiments, the anti-PD-L1 antibody or conjugate does not comprise a functional Fc region. Non-functional Fc regions include non-existent or truncated Fc regions, or Fc regions containing amino acid substitutions or deletions that reduce or eliminate Fc-mediated activity, such as effector functions (e.g., ADCC or ADCP activity). In some embodiments, the Fc region is engineered to exhibit ADCC and / or ADCP activity or engineered to exhibit enhanced effector functions, such as enhanced ADCC and / or ADCP activity. The Fc region modification that causes enhanced ADCC and / or ADCP activity may be any known modification or may be determined empirically. Exemplary modifications include, but are not limited to, modifications to the Fc region described herein. In some embodiments, the antibody is produced so that it is defucosylated.

[0321] In some aspects, conjugates of the provided anti-PD-L1 antibodies or antibody fragments with one or more toxic, cytotoxic, or fluorescent agents can be prepared using any of a variety of known protein coupling agents (e.g., linkers) (see Vitetta et al., Science 238:1098 (1987), WO94 / 11026). In some embodiments, the linker is a peptide or polypeptide or a chemical linker. In some embodiments, the linker is a releasable linker or a cleavable linker. The linker can be a "cleavable linker" or "releasable linker" that facilitates the release of the cytotoxic drug in the cell, such as an acid-labile linker, a peptidase-sensitive linker, a photolabile linker, a dimethyl linker, and a disulfide-containing linker (Chari et al., Cancer Res. 52:127-131 (1992); U.S. Pat. No. 5,208,020). In some embodiments, the releasable linker or cleavable linker is released or cleaved in the presence of one or more conditions or factors present in the tumor microenvironment (TME), including matrix metalloproteinases (MMPs), hypoxic conditions, or acidic conditions.

[0322] III. Treatment Methods and Uses of Anti-PD-L1 Antibodies and Conjugates

[0323] In some embodiments, provided anti-PD-L1 antibodies, fragments or conjugates are used in methods of treating lesions or tumors and uses. In some embodiments, provided methods and uses involve administering an anti-PD-L1 antibody or fragment or conjugate. In a specific embodiment, the conjugate contains a photoactivatable phthalocyanine dye, and the methods and uses involve administering the conjugate and irradiating the target area with light of a wavelength suitable for the phthalocyanine dye, such that the light excites the dye and kills cells expressing PD-L1 on their surface, for example, as described herein. Any of such methods and uses results in enhancing, activating, inducing, eliciting, strengthening or supporting immune function, such as local and / or systemic immunity; reducing or eliminating lesions (e.g., tumors); reducing or inhibiting tumor growth; reducing, inhibiting or eliminating tumor cell metastasis, or any combination thereof.

[0324] However, in some aspects, the compositions, methods, and uses provided can further enhance, activate, induce, elicit, strengthen, or support immune functions, such as local and / or systemic immunity, by inhibiting PD-1:PD-L1 interactions and / or killing and eliminating cells expressing PD-L1 on the cell surface. Thus, eliminating or killing cells expressing PD-L1 on the surface, particularly immune cells with immunosuppressive functions, such as M2 tumor-associated macrophages (M2 TAMs), M1 or M2 macrophages, tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs), can be used to enhance, activate, induce, elicit, strengthen, or support immune functions, such as local and / or systemic immunity, such as anti-tumor or anti-cancer immunity. Elimination or killing of cells expressing PD-L1 on the surface can be achieved through: Fc-mediated effector functions; delivery of toxic payloads to cells expressing PD-L1; and / or after irradiating the target area with light of a wavelength suitable for photoactivatable dyes, such that the dye is photoexcited and cell killing occurs.

[0325] In some aspects, the methods and uses provided can enhance, activate, induce, recruit or support lymphocyte infiltration into a tumor or lesion. In some embodiments, the methods and uses provided activate innate responses in tumors, such that the activation of dendritic cells (e.g., activated dendritic cells) in tumors increases. In some aspects, the methods and uses provided activate adaptive immune responses, such that the infiltration of CD8+T cells increases. In some embodiments, the methods and uses provided reduce the number of CD8+T cells exhausted in tumors. In some embodiments, the methods and uses provided increase the intratumoral infiltration of newly activated CD8+T cells. In some embodiments, the methods and uses provided can cause improvements in therapeutic effects, such as by selecting non-exhausted effector cells with higher content or quantity, such as individuals with CD8+T cells for treatment, and / or by improving the activity or reaction of non-exhausted effector cells.

[0326] In some aspects, the compositions, methods, and uses provided can be used for tumors that are resistant to checkpoint inhibitor immunotherapy or difficult to treat with such immunotherapy. In some aspects, the compositions, methods, and uses provided can be used for tumors that are resistant to anti-PD-L1 immunotherapy or difficult to treat with such immunotherapy. Tumors, such as solid tumors, can develop resistance to checkpoint inhibitor immunotherapy through several mechanisms, including but not limited to irreversible T cell exhaustion; insufficient T cell activation; tumor cell immunoediting that causes upregulation of compensatory inhibitory signaling of T cells; generation of an immunosuppressive tumor microenvironment, such as through increased infiltration of Tregs, MDSCs, tumor-associated macrophages (e.g., M2 macrophages); increased levels of tumor-derived cytokines and chemokines (e.g., TGF-β, CXCL8); silencing of Th1-type chemokines; indoleamine 2,3-dioxygenase (IDO) production; and excess extracellular adenosine. The compositions, methods, and uses provided can be used to overcome one or more checkpoint inhibitor resistance mechanisms employed by some tumors.

[0327] In addition, administration of antibodies, cytotoxic conjugates, or photoactivatable conjugates followed by irradiation can also directly kill cancer cells expressing PD-L1, thereby inhibiting or reducing tumor growth. PD-L1 antibodies, cytotoxic conjugates, or photoactivatable conjugates, such as anti-PD-L1-phthalocyanine conjugates, can directly or indirectly affect and kill tumor cells or cells present in a tumor or a tumor's microenvironment (also referred to as the tumor microenvironment; TME), including tumor cells in a different location from the primary tumor, metastatic tumors, newly generated tumor cells, and / or tumors expressing different types or cell surface antigens. Thus, the provided compositions, methods, and uses can provide effective treatment even for tumor cells that do not express cell surface PD-L1, have low or substantially no response to previous therapy, such as previous immunomodulator therapy, have failed previous therapy, have relapsed after previous therapy, are difficult to treat with previous therapy, and / or are resistant to previous therapy. In certain embodiments, provided compositions, methods, and uses can treat tumors or lesions that are unresponsive, resistant, or difficult to treat with anti-PD-L1, anti-PD-1, and / or anti-CTLA-4 therapy.

[0328] In some embodiments, the compositions, methods and uses provided herein are also effective in treating larger tumors and exhibiting greater immunosuppressive effects than smaller tumors. Such tumors may be less responsive or unresponsive to other treatments, such as with immunomodulators, such as treatments with immune checkpoint inhibitors (e.g., anti-PD-L1, anti-PD-1 and / or anti-CTLA-4 therapies). In such cases, anti-PD-L1 photoimmunotherapy provided by administering an anti-PD-L1 conjugate described herein and subsequent irradiation can effectively inhibit or substantially reduce the growth of larger tumors, which in some cases cannot be effectively inhibited by other immunomodulator therapies and / or anti-cancer therapies. In some embodiments, the compositions, methods and uses provided herein are effective in treating tumors that are larger in size and resistant to anti-PD-L1, anti-PD-1 and / or anti-CTLA-4 therapies.

[0329] In some aspects, methods and uses are provided for treating tumors or lesions of an individual via activation of immune cell responses. Immune cell activation can be direct or indirect activation. In some aspects, methods and uses are provided for treating individuals who have a low response to, no response to, drug resistance to, difficult to treat with, unable to respond to, or relapsed after a previous immunotherapy for a tumor or lesion (e.g., anti-PD-L1, anti-PD-1, and / or anti-CTLA-4 therapy).

[0330] In some aspects, the method involves administering to an individual having a tumor or lesion a conjugate comprising a phthalocyanine dye (such as IR700) attached to an anti-PD-L1 antibody described herein. In some aspects, the method also ... at a wavelength of at or about 600 nm to at or about 850 nm and at a rate of from 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 Or irradiating cells expressing PD-L1, such as target areas where immune cells expressing PD-L1 are located, at a dose of from 2 J / cm fiber length or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length, thereby causing the killing of cells expressing PD-L1 and thereby inhibiting the growth of the tumor or lesion. In some aspects, the method can cause the killing of cells expressing PD-L1 and thereby increase the number or activity of immune cells in the tumor or lesion and / or in the microenvironment of the tumor or lesion.

[0331] In some embodiments, methods and uses are provided for treating a tumor or lesion in a subject by activating an immune cell response in a subject having a tumor or lesion who has been administered a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds PD-L1, comprising administering a dose of 25 J / cm2 of a phthalocyanine dye at a wavelength of at or about 600 nm to at or about 850 nm. 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 Or irradiating a target area where immune cells expressing PD-L1 are located with a dose of from 2 J / cm of fiber length or about 2 J / cm of fiber length to 500 J / cm of fiber length or about 500 J / cm of fiber length; wherein the method results in killing of the PD-L1-expressing cells and thereby inhibiting the growth of the tumor or lesion. In some embodiments, the PD-L1-expressing cells are immune cells. In some embodiments, the PD-L1-expressing cells are tumor cells.

[0332] In some embodiments, methods and uses are provided for treating an individual who has had a low response to, is unresponsive to, is resistant to, is refractory to, fails to respond to, or has relapsed after a previous immunotherapy for a tumor or lesion, comprising administering a dose of 25 J / cm2 at a wavelength of 600 nm or about 600 nm to 850 nm or about 850 nm at a dose of 25 J / cm2 to 850 nm. 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 or irradiating a target area of ​​immune cells expressing PD-L1 in an individual with a tumor or lesion that has had a low response or no response to previous immunotherapy with a dose of from 2 J / cm fiber length or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length, wherein the method results in killing of cells expressing PD-L1 and thereby increases the number or activity of immune cells in the tumor and / or in the tumor microenvironment. In some of any of the embodiments, the cells expressing PD-L1 are immune cells.

[0333] In some embodiments, methods and uses are provided for enhancing the response of an individual with a tumor or lesion to an anticancer agent. In some aspects, the method involves administering an anticancer agent to an individual with a tumor or lesion. In some aspects, a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds PD-L1 is administered to the individual. In some aspects, the dose is from 25 J / cm2 at a wavelength of from about 600 nm to about 850 nm. 2or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 Or irradiating the target area where immune cells expressing PD-L1 are located with a dose of from 2 J / cm fiber length or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length causes a growth inhibition of the tumor or the lesion that is greater than the inhibition caused by treatment with the anticancer agent alone.

[0334] In some embodiments, methods and uses are provided for enhancing the response of an individual having a tumor or lesion to an anticancer agent, comprising: administering to the individual a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1; and administering to the individual a phthalocyanine dye at a wavelength of at or about 600 nm to at or about 850 nm at a dose of from 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 or irradiating a target area where immune cells expressing PD-L1 are located with a dose of from 2 J / cm fiber length or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length, wherein the individual has been administered an anti-cancer agent; and causing an inhibition of growth of the tumor or the lesion that is greater than that caused by treatment with the anti-cancer agent alone.

[0335] In some embodiments, methods and uses are provided for enhancing the response of an individual having a tumor or lesion to an anticancer agent, comprising: administering an anticancer agent to the individual; wherein the individual has been receiving a treatment comprising administering to the individual a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1; and administering to the individual a dose of 25 J / cm at a wavelength of at or about 600 nm to at or about 850 nm. 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 Or irradiating the target area where immune cells expressing PD-L1 are located with a dose of from 2 J / cm fiber length or about 2 J / cm fiber length to 500 J / cm fiber length or about 500 J / cm fiber length, and wherein the administration of the anticancer agent and the treatment cause a growth inhibition of the tumor or the lesion that is greater than the inhibition caused by the anticancer agent alone.

[0336] In some embodiments, methods and uses of vaccinating or immunizing an individual to generate an anti-cancer immune response are provided. In some aspects, vaccinating or immunizing an individual to generate an anti-cancer immune response can inhibit the growth and / or reduce the size of a first tumor or lesion; and delay or prevent the emergence, growth or establishment of one or more second tumors or lesions, for example, located distal to the treated first tumor or lesion. In some aspects, the method involves administering to the individual a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds PD-L1. In some aspects, the method involves irradiating the target area to elicit an anti-cancer response selected from the delay or inhibition of the growth of a tumor that appears in the individual, or the emergence or increase of T memory cells near the tumor.

[0337] In some embodiments, methods and uses are provided for vaccinating or immunizing an individual to generate an anti-cancer immune response, comprising: irradiating a target area in an individual to whom a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds PD-L1 has been administered; and eliciting an anti-cancer response selected from the group consisting of a delay or inhibition of tumor appearance or growth, or the appearance or increase of T memory cells in the vicinity of the tumor in the individual.

[0338] In some embodiments, one or more steps of the method are repeated. In some embodiments, the administration of the conjugate is repeated one or more times, optionally wherein after each repeated administration of the conjugate, the irradiation step is repeated. In some embodiments, further comprising administering an additional therapeutic agent or anti-cancer therapy.

[0339] A. Methods for Stimulating or Enhancing Anti-Cancer Immune Responses

[0340] In some aspects, provided methods and uses employing compositions comprising anti-PD-L1 conjugates can result in an enhanced immune response, such as a systemic and / or local immune response, in the individual, which in turn can result in an enhanced response to therapy or treatment for a tumor, lesion, or cancer. In some aspects, the methods and uses herein comprise administering an anti-PD-L1 conjugate to the individual, and following administration of the conjugate, irradiating a target area, such as a target area where cells expressing PD-L1 are present, e.g., a tumor, near a tumor, a lymph node, near a lymph node, or in the tumor microenvironment.

[0341] In some aspects, the embodiments provided can stimulate, enhance, activate, induce, cause, strengthen, enhance or support an immune response, such as a systemic immune response, in an individual suffering from a tumor, lesion or cancer. In some embodiments, the methods and uses provided enhance the systemic immune response of an individual suffering from a tumor, lesion or cancer. "Systemic immune response" refers to the ability of an individual's immune system to respond in a systemic manner to one or more immune attacks, including immune attacks associated with tumors, lesions or cancer. A systemic immune response may include a systemic response of an individual's acquired immune system and / or innate immune system. A systemic immune response may include an anti-tumor or anti-cancer response generated by an individual's acquired immune system and / or innate immune system. In some aspects, a systemic immune response includes immune responses in different tissues, including the bloodstream, lymph nodes, bone marrow, spleen and / or tumor microenvironment, and in some cases, includes a coordinated response of tissues and organs and various cells and factors of tissues and organs. In some embodiments, the embodiments provided can stimulate, enhance, activate, induce, cause, strengthen, enhance or support an anti-cancer or anti-tumor immune response of an individual's own immune system, including the acquired immune system and / or innate immune system. In some aspects, provided methods and uses result in an enhanced innate immune response in the individual.

[0342] In some aspects, provided embodiments can also exhibit abscopal effects. In some aspects, "abscopal effects" refer to a therapeutic effect in which tumors not directly treated or distant from the local treatment site, such as distal or metastatic tumors, are also treated, such as a reduction in tumor size.

[0343] In some aspects, the embodiments provided can achieve tumor immunity. In these aspects, the embodiments provided prevent or hinder the growth of new tumors or metastases. In some embodiments, the tumor growth inhibition achieved by the embodiments provided results in a durable anti-tumor response. In some embodiments, the tumor growth inhibition achieved by the embodiments provided prolongs progression-free survival. In some embodiments, the tumor growth inhibition achieved by the embodiments provided reduces the chance of recurrence and / or reduces the chance of metastasis. In some aspects, the embodiments provided can achieve immunity to the same tumor type or different tumor types in the treated individual. In some aspects, the embodiments provided can inhibit the growth of tumors from different tumor lineages, i.e., different types of tumors that have or may develop in the treated individual.

[0344] In some aspects, the target area is an area comprising cells expressing PD-L1. In some embodiments, the cells expressing PD-L1 are immune cells. In some of any embodiments, the method causes the killing of cells expressing PD-L1, such as immune cells expressing PD-L1. In some embodiments, the immune cells expressing PD-L1 are selected from the following groups: monocytes, macrophages, dendritic cells (DC), M2 tumor-associated macrophages (M2 TAM), M1 or M2 macrophages, tolerogenic dendritic cells (tDC) and myeloid-derived suppressor cells (MDSC). In some embodiments, the immune cells expressing PD-L1 are located in tumors, tumor microenvironment or lymph nodes.

[0345] In some aspects, the target area irradiated according to the provided embodiments is a tumor, such as a primary tumor, near a tumor (such as a primary tumor), or a tumor microenvironment (TME). In some embodiments, the target area is close to a tumor or adjacent to a tumor, or near a tumor or tumor cells. In some embodiments, the target area is a tumor. In some embodiments, the target area is a primary tumor. In some embodiments, the target area is a secondary tumor or a metastatic tumor. In some embodiments, the target area is a tumor microenvironment.

[0346] In some embodiments, the target area is a lymph node or is near a lymph node. In some embodiments, the target area is a lymph node, such as a lymph node containing cells expressing PD-L1, or is near such a lymph node. In some embodiments, the target area is a lymph node. In some embodiments, the target area is near a lymph node.

[0347] In some aspects, provided embodiments can stimulate or enhance a systemic response, such as a systemic immune response, against one or more primary tumors or lesions and / or one or more secondary tumors or lesions, such as metastatic tumors or lesions, or tumors or lesions of different types.

[0348] In some aspects, the embodiments provided stimulate or enhance an individual's immune response, such as an individual's anti-cancer immune response, by removing immune cells expressing PD-L1, such as cells that may have immunosuppressive function, such as monocytes, macrophages (such as M1 macrophages, M2 macrophages and / or M2 tumor-associated macrophages (M2 TAMs)), dendritic cells (DCs), tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs) in some cases. In some aspects, the embodiments provided stimulate or enhance an individual's immune response, such as a systemic and / or local immune response targeting a tumor, lesion, or cancer, by killing and eliminating immunosuppressive cells expressing PD-L1, such as M2 TAMs, tDCs, or MDSCs. As exemplified herein, tumor growth inhibition following administration of a PD-L1-phthalocyanine dye conjugate and light irradiation requires the presence and / or activity of the individual's CD8+ T cells, because depletion of the individual's CD8+ T cells results in tumor growth similar to that in controls administered saline. In some aspects, immunosuppressive cells, such as M2 TAMs, tDCs, or MDSCs, such as cells expressing PD-L1, suppress the function and / or activity of individual immune cells, such as CD8+ T cells or natural killer (NK) cells. By killing and eliminating immunosuppressive cells, such as immunosuppressive cells expressing PD-L1, including M2 TAMs, tDCs, or MDSCs, the provided embodiments can stimulate and enhance the immune response of an individual. As exemplified herein, such treatments according to the provided embodiments result in growth inhibition of one or more primary tumors, such as complete responses to the treatment, and growth inhibition of one or more second tumors from the primary tumor or lesion, such as second tumors of the same or different type and / or origin and / or present at different sites, such as at distal sites.

[0349] In some aspects, the growth inhibition of the tumor or the lesion and / or the killing of the cells expressing PD-L1 depends on the presence of CD8+T cells. In some embodiments, before the administration, the individual has a tumor or lesion with a small number or amount of CD8+T cell infiltration. In some embodiments, after the administration and the irradiation, the number, amount or activity of immune cells in the tumor or in the tumor microenvironment increases. In some embodiments, after the administration and the irradiation, the number or amount of CD8+T cell infiltration in the tumor or the lesion increases. In some embodiments, after the administration and the irradiation, the number of memory T cells near the tumor increases.

[0350] In some aspects, the stimulated or enhanced systemic immune response includes systemic CD8 +Increase in the number and / or activity of T effector cells; increase in systemic T cell cytotoxicity against tumor cells as measured using CTL assays using cells from the spleen, peripheral blood, bone marrow, or lymph nodes; intratumoral CD8 + Increased number, activity, and / or activation of T effector cells; systemic CD8 + an increase in T cell activation; an increase in systemic dendritic cell activation; an increase in dendritic cell activation in a primary or secondary (e.g., metastatic or new) tumor or lesion; an increase in intratumoral dendritic cell infiltration in a primary or secondary (e.g., metastatic or new) tumor or lesion; an increase in new T cell activation in a primary or secondary (e.g., metastatic or new) tumor or lesion; an increase in T cell diversity in a primary or secondary (e.g., metastatic or new) tumor or lesion; a decrease in systemic regulatory T cells; a decrease in regulatory T cells in a primary or secondary (e.g., metastatic or new) tumor or lesion; a decrease in systemic myeloid-derived suppressor cells; a decrease in intratumoral myeloid-derived suppressor cells in a primary or secondary (e.g., metastatic or new) tumor or lesion; a decrease in tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs) in a primary or secondary (e.g., metastatic or new) tumor or lesion; or any combination thereof. In some cases, systemic response can be assessed by sampling blood, tissue, cells or other fluids from the individual and assessing increases in proinflammatory cytokines, immune cell activation markers and / or T cell diversity. In some aspects, systemic response can be assessed by analyzing cells directly or indirectly affected by the method. For example, cells can be collected from the individual between day 4 and day 28 after treatment or at any time after the step of irradiating the individual's primary tumor.

[0351] In some aspects, the embodiments provided can stimulate, enhance, strengthen, strengthen or support an immune response, such as a local reaction, such as a local immune response, in an individual suffering from a tumor, lesion or cancer. In some embodiments, the methods and uses provided enhance the local reaction of an individual suffering from a tumor, lesion or cancer. A "local immune response" refers to an immune response in a tissue or organ to one or more immune attacks, including immune attacks associated with a tumor, lesion or cancer. The local immune response may include the acquired immune system and / or the innate immune system. In some aspects, local immunity includes immune responses that occur simultaneously in different tissues, such as the bloodstream, lymph nodes, bone marrow, spleen and / or tumor microenvironment.

[0352] In some aspects, the stimulated or enhanced local immune response includes intratumoral CD8 + T effector cells (e.g., CD3 + CD8 +cells) increased in number and / or activity, CD8 + Increased activation of T effector cells, dendritic cells in tumors (CD11c + ) cell infiltration, activation of dendritic cells in tumors (e.g., CD11c + CD80 + and / or CD11c + CD40 + ) increased, intratumoral antigen-presenting dendritic cells (CD11b + CD103 + CD11c + ) increase, activation of new T cells in tumors (e.g. CD3 + CD8 + PD1 - cells), increased T cell diversity within the tumor, and increased neutrophils within the tumor (CD11b + Cy6C - / 低 Ly6G + cells), increased intratumoral macrophages (e.g., CD11b + f / 4 / 80 + cells), decreased regulatory T cells (Tregs) within the tumor, and decreased myeloid-derived suppressor cells (MDSCs; e.g., CD11b + Ly6C + Ly6G - cells), a decrease in tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs) within the tumor, and exhausted T cells within the tumor, such as exhausted CD8+ T cells (e.g., PD-1 + CTLA-4 + CD3 + CD8 + In some aspects, the cell surface phenotype of the cell, such as the immune cells indicating a local immune response or an innate immune response, is assessed by staining with a labeled antibody using a reagent that can be used to detect marker expression on the surface. In some aspects, the cell surface phenotype of the cell, such as the immune cells indicating a local immune response or an innate immune response, is assessed by staining with a labeled antibody using a reagent that can be used to detect marker expression on the surface. In some aspects, the cell surface phenotype of the cell, such as the immune cells indicating a local immune response or an innate immune response, is detected using flow cytometry.

[0353] In some cases, local reactions, such as local immune reactions, can be assessed by obtaining blood, tissue, or other samples from the individual and assessing the increase in anti-immune cell types in the tumor or TME and / or assessing the increase or appearance of local immune activation markers. In some aspects, local reactions, such as local immune reactions, can be assessed by analyzing cells directly or indirectly affected by the method. For example, cells can be collected from the individual between day 4 and day 28 after treatment or at any time after the step of irradiating the individual's primary tumor.

[0354] In some aspects, the methods and uses also involve administering an additional therapeutic agent, such as an immunomodulator, for example, an immune checkpoint inhibitor. The immunomodulator can be administered before, simultaneously with, or after administering the conjugate. In some aspects, administering the additional therapeutic agent, such as an immunomodulator, can also cause stimulation, enhancement, activation, induction, strengthening, or support of an immune response, such as a systemic and / or local immune response of an individual, including an anticancer or antitumor response. Exemplary additional therapeutic agents, compositions, combinations, methods, and uses include those described herein, such as those described in Section V.

[0355] B. Tumors and Lesions for Anti-PD-L1 Conjugate Therapy

[0356] The methods described herein include administering an anti-PD-L1 conjugate and irradiating a target area in the subject's body with a wavelength of light that activates the phthalocyanine dye portion of the conjugate, such as a tumor or lesion, near a tumor, a lymph node, near a lymph node, or a tumor microenvironment (TME) of a tumor lesion to achieve, for example, cell killing of cells expressing PD-L1 on the surface. In some embodiments, the methods and uses provided herein include treating an individual with one or more tumors or lesions, such as one or more primary tumors or lesions (or first tumors or lesions), one or more secondary tumors or lesions (or second tumors or lesions), one or more newly generated tumors or lesions, and / or one or more metastatic tumors or lesions. An individual may have one, two, three or more tumors. Such tumors may be in one or more tissues or organs, such as in one tissue or organ, in two different tissues or organs, in three different tissues or organs, or more different tissues or organs. In some aspects, one or more of the tumors to be treated express PD-L1 on the surface of the cells contained in the tumor. In some aspects, one or more of the tumors to be treated contain, consist primarily of, have a greater number of, or consist entirely of cells that do not express PD-L1, have low PD-L1 expression, or are PD-L1 negative. In some aspects, one or more of the tumors to be treated contain, consist primarily of, have a greater number of, or consist entirely of cells that have a reduced response to, are resistant to, or become resistant to (i.e., acquired resistance to) PD1 / PD-L1 checkpoint blockade.

[0357] In some aspects, the tumor or lesion treated according to the embodiments provided is not treated with an immune checkpoint inhibitor, or has not previously been treated with an immune checkpoint inhibitor, such as one or more anti-PD-1, anti-PD-L1 and / or anti-CTLA-4 therapies. In some embodiments, the tumor or lesion has not been treated with anti-PD-1 therapy (has not been treated with such therapy). In some embodiments, the tumor or lesion has not been treated with anti-PD-L1 therapy (has not been treated with such therapy). In some embodiments, the tumor or lesion has not been treated with anti-CTLA-4 therapy (has not been treated with such therapy). In some embodiments, the individual with the tumor or lesion to be treated according to the embodiments provided is an individual who has not been treated with an immune checkpoint inhibitor. In some embodiments, the individual to be treated is an individual who has not been treated with anti-PD-1 therapy. In some embodiments, the individual to be treated is an individual who has not been treated with anti-PD-L1 therapy. In some embodiments, the individual to be treated is an individual who has not been treated with anti-CTLA-4 therapy. Treatment of an individual with immune checkpoint inhibitors, such as anti-PD-1 antibodies, can cause depletion of CD8+ effector T cells in, around, and / or throughout the tumor. This can prevent CD8+ T cells from recognizing and locating to the tumor, or it can cause CD8+ T cells to be located to the tumor or near the tumor, but still inefficient, resulting in resistance to checkpoint inhibitors (e.g., PD-1 / PD-L1). Therefore, in some cases, inefficient or insufficient CD8+ effector T cell activity can be alleviated by avoiding immune checkpoint inhibitor therapy (e.g., anti-PD-1, anti-PD-L1, and / or anti-CTLA-4 therapy) before adopting the provided compositions, methods, or uses. In some embodiments, the response of the tumor or lesion to the treatment herein is achieved by administering an anti-PD-L1 conjugate to treat the tumor or lesion and subsequently irradiating the tumor or lesion before any treatment with an immune checkpoint inhibitor, such as PD-1, PD-L1, and / or CTLA-4 directed therapy (such as anti-PD-1 antibodies and anti-PD-L1 antibodies, and / or anti-CTLA-4 antibodies). In some embodiments, the method of treatment comprises selecting an individual who has not been treated with immune checkpoint inhibitor (e.g., anti-PD-1, anti-PD-L1, and / or anti-CTLA-4) therapy, and treating such individual (i.e., such individual's tumor or lesion) with an anti-PD-L1 conjugate followed by irradiation.

[0358] In some aspects, the tumor or lesion treated according to the provided embodiments is associated with a cancer selected from the group consisting of colorectal cancer, colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell carcinoma, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, small intestine cancer, spindle cell neoplasm, liver cancer, liver cancer, peripheral nerve cancer, brain cancer, skeletal muscle cancer, smooth muscle cancer, bone cancer, adipose tissue cancer, cervical cancer, uterine cancer, genital cancer, lymphoma, and multiple myeloma.

[0359] In some aspects, the tumor or lesion treated according to the provided embodiment includes one or more primary (e.g., first) tumors or lesions. In some aspects, a primary tumor or lesion may include the first or primary tumor or lesion in an individual. In some aspects, the individual may have one or more primary tumors or lesions. In some embodiments, one or more primary tumors may be one or more solid tumors, lymphomas, or leukemias. The tumor may be a tumor of the lung, stomach, liver, pancreas, breast, esophagus, head and neck, brain, peripheral nerves, skin, small intestine, large intestine, rectum, anus, ovary, uterus, bladder, prostate, adipose tissue, skeletal muscle, smooth muscle, blood vessel, bone, bone marrow, eyes, tongue, lymph nodes, spleen, kidney, cervix uteri, male genitalia, female genitalia, testicles, or a tumor of unknown initial origin.

[0360] In some aspects, the tumor or lesion treated according to the provided embodiments includes one or more second tumors or lesions, such as metastatic tumors or lesions, or newly generated tumors or lesions. In some aspects, the one or more second tumors or lesions are derived from the metastasis of the first tumor or lesion. In some embodiments, the one or more second tumors or lesions are tumors that are not derived from the metastasis of the first tumor or lesion. In some aspects, the one or more second tumors or lesions are phenotypically and / or genotypically different from the first tumor or lesion. In some aspects, the one or more second tumors or lesions are phenotypically different from the first tumor or lesion. In some aspects, the one or more second tumors or lesions are genotypically different from the first tumor or lesion. In some aspects, the one or more second tumors or lesions are newly generated tumors or lesions. In some aspects, the one or more second tumors or lesions are from a source different from the first tumor or lesion. In some aspects, the one or more second tumors or lesions are produced by an organ or cell different from the first tumor or lesion. In some embodiments, the one or more second tumors or lesions can be one or more solid tumors, lymphomas, or leukemias. The one or more second tumors or lesions can be lung, stomach, liver, pancreas, breast, esophagus, head and neck, brain, peripheral nerves, skin, small intestine, large intestine, rectum, anus, ovary, uterus, bladder, prostate, adipose tissue, skeletal muscle, smooth muscle, blood vessels, bone, bone marrow, eye, tongue, lymph node, spleen, kidney, cervix, male genitalia, female genitalia, testicles, or tumors of unknown origin.

[0361] In some embodiments, immunity against a second tumor or lesion is achieved when the first tumor is irradiated after administration of the provided anti-PD-L1 conjugate, and the volume of the first tumor is reduced. In such embodiments, the volume of the first tumor is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the volume of the first tumor is reduced by at least 50% or at least about 50%. In some embodiments, the volume of the first tumor is reduced by at least 75% or at least about 75%. In some embodiments, immunity against a second tumor or lesion is achieved after treatment of the first tumor when the first tumor achieves a partial or complete response (PR or CR). In some embodiments, immunity against a second tumor or lesion is achieved after treatment of the first tumor when the first tumor achieves a CR.

[0362] In some cases, the target area for irradiation may be a primary tumor or lesion, or in the vicinity of the primary tumor or lesion. In other cases, the target area for irradiation is not the primary tumor or lesion, but rather a different area where cells expressing PD-L1 are present, such as a lymph node, or a secondary tumor or lesion, or in the vicinity of a secondary tumor or lesion.

[0363] In some aspects, after treatment with anti-PD-L1 conjugates and photoirradiation according to the embodiments provided, the growth of one or more primary tumors or lesions is inhibited, the volume of one or more primary tumors or lesions is reduced, or both tumor growth and volume are reduced. In some aspects, after treatment with anti-PD-L1 conjugates and photoirradiation according to the embodiments provided, the growth of one or more secondary or metastatic tumors or lesions is inhibited, the volume of one or more secondary or metastatic tumors or lesions is reduced, and both tumor growth and volume are reduced. In some aspects, treatment according to the embodiments provided delays the regrowth of tumors or lesions, prevents the recurrence of cancer (such as cancer associated with the tumor or lesion) or prolongs the duration of cancer remission, prevents or inhibits the generation and / or growth of one or more second tumors or lesions, including the generation and / or growth of a second tumor or lesion of a different type from the primary tumor or lesion, and / or prevents or inhibits the generation and / or growth of metastasis.

[0364] In some embodiments, the subject is administered an anti-PD-L1 conjugate to treat a first tumor or a first lesion and / or inhibit its growth; and the method inhibits or delays the appearance of one or more second tumors or lesions, or metastasis of the first tumor or the first lesion.

[0365] In some aspects, primary tumors or lesions contain cells expressing PD-L1 on the surface. In some aspects, cells expressing PD-L1 are immune cells, such as immunosuppressive cells, for example, M2TAM, tDC or MDSC. In some aspects, cells expressing PD-L1 are tumor-associated fibroblasts or cancer-associated fibroblasts (CAF). In some cases, cells expressing PD-L1 are tumor cells or cancer cells. In some of any embodiments, the individual to be treated has one or more cells expressing PD-L1, such as one or more cells expressing PD-L1 associated with the tumor, lesion or cancer.

[0366] In some embodiments, the tumor, lesion or cancer to be treated contains tumor or cancer cells that do not express PD-L1. In some embodiments, the tumor or the lesion comprises PD-L1 negative tumor cells. In some embodiments, more than 40%, 50%, 60%, 70%, 80%, 90% or 95% or more than about 40%, 50%, 60%, 70%, 80%, 90% or 95% of the tumor cells in the tumor or the lesion are PD-L1 negative tumor cells. In some aspects, PD-L1 negative tumor cells may refer to tumor cells that do not express a detectable amount of PD-L1 on the surface or tumor cells whose expression of PD-L1 is below a threshold, such as a detectable threshold. In some embodiments, PD-L1 negative tumor cells include tumor cells that are not specifically recognized by anti-PD-L1 antibodies. In some cases, the expression of PD-L1 is determined by flow cytometry. In some aspects, provided embodiments result in indirect killing of tumor cells or cancer cells, such as by eliminating immunosuppressive cells, e.g., M2 TAMs, tDCs, or MDSCs, and enhancing the function and / or activity of effector cells in the immune system, such as CD8+ T cells, so that they can exert an anti-tumor or anti-cancer response to eliminate tumor cells or cancer cells.

[0367] In some embodiments, the tumor, lesion, or cancer to be treated contains tumor or cancer cells that express PD-L1. In some aspects, administration of an anti-PD-L1 conjugate followed by light irradiation can directly kill cells that express PD-L1. In some aspects, provided embodiments result in direct killing of tumor cells that express PD-L1.

[0368] In some embodiments, the methods and uses provided herein include treating individuals with invasive tumor cells, such as when cells derived from a primary tumor invade into surrounding tissues. The method includes administering an anti-PD-L1 conjugate to an individual with invasive tumor cells and, after administering the conjugate, irradiating the target area with a wavelength suitable for the selected phthalocyanine dye. In some embodiments, the method includes administering an immunomodulator, such as an immune checkpoint inhibitor, before, simultaneously with, or after administering the conjugate. In some aspects, invasive tumor cells refer to cells derived from a primary tumor and have invaded into the same organ as the primary tumor or adjacent organs or surrounding tissues of a body cavity in an individual with a primary tumor.

[0369] In some cases, the methods and uses provided herein include irradiating a target area. In some aspects, the target area is one or more primary tumors, and some or all of the invasive tumor cells are not irradiated, and in such methods, the growth of the invasive tumor cells is inhibited, reduced, or eliminated, the volume of the one or more invasive tumors is reduced, or any combination thereof. In some embodiments, the growth of the primary tumor is also inhibited, reduced, or eliminated, and the volume of the one or more primary tumors is also reduced along with the effect on the one or more invasive tumor cells.

[0370] In some embodiments, the invasive tumor cells are contained within a solid tumor. In some embodiments, the invasive tumor cells are contained within a body fluid, including but not limited to peritoneal fluid, pleural fluid, and cerebrospinal fluid. In some embodiments, the invasive tumor cells are contained within one or more body cavity effusions, including but not limited to peritoneal effusions (ascites), pleural effusions, and pericardial effusions.

[0371] In some embodiments, the methods and uses provided herein include treating an individual having one or more primary tumors and metastatic tumor cells. The method comprises administering an anti-PD-L1 conjugate to an individual having a primary tumor and metastatic tumor cells and, after administering the conjugate, irradiating the target area with a wavelength appropriate for the selected phthalocyanine dye. In such methods, the growth of metastatic tumor cells is inhibited, reduced, or eliminated, the size of one or more metastatic tumors is reduced, or any combination thereof.

[0372] In some embodiments of the methods and uses provided herein, the metastatic tumor cells are distal to the primary tumor and some or all of the metastatic tumor cells are not irradiated, eg, not directly irradiated.

[0373] In some embodiments of the methods and uses, only the target area, such as a target area containing a lymph node or primary tumor or lesion and / or a target area near a lymph node or primary tumor or lesion, is irradiated. In some aspects, a second tumor or lesion, such as a metastatic tumor or lesion, is not irradiated.

[0374] In some aspects, metastatic tumor cells include cells derived from a primary tumor and spread to distal tissues or organs or cells derived from distal tissues or organs in a subject with the primary tumor. The metastatic tumor cells may be located in one or more locations in the lungs, stomach, liver, pancreas, breast, esophagus, head and neck, brain, peripheral nerves, skin, small intestine, large intestine, rectum, anus, ovary, uterus, bladder, prostate, adipose tissue, skeletal muscle, smooth muscle, blood vessel, bone, bone marrow, eyes, tongue, lymph nodes, spleen, kidney, cervix uteri, male genitalia, female genitalia, testicles, blood, bone marrow, cerebrospinal fluid, or any other tissue or organ. In some embodiments, metastatic tumor cells are contained in solid tumors. In some embodiments, metastatic tumor cells are circulating tumor cells or are unrelated to a mass.

[0375] In some embodiments, the methods and uses comprise administering an immunomodulatory agent, such as a checkpoint inhibitor, prior to, concurrently with, or after the administration of the conjugate. In some embodiments, the methods and uses comprise administering a second conjugate, such as a second immunoconjugate, followed by irradiation concurrently with, prior to, or after the administration of the just-provided conjugate. In some embodiments, the methods and uses comprise administering one or more additional anti-cancer therapies, such as chemotherapy, anti-angiogenic therapy, kinase inhibitors, radiation therapy, small molecule therapy, or other therapies, such as any of the therapies described herein under the section entitled "Combination Therapies."

[0376] C. Methods and Compositions for Treating Tumors or Tumor Cells That Are Less Responsive, Refractory, or Non-Responsive to Previous Therapeutic Treatments

[0377] In some embodiments, compositions containing an anti-PD-L1 conjugate, i.e., a phthalocyanine dye-targeting molecule conjugate, wherein the targeting molecule binds to PD-L1 (e.g., an anti-PD-L1 antibody-IR700 conjugate), and methods and uses thereof are provided for the treatment or therapy of a tumor or cancer that has not been treated with one or more prior therapies, such as immunomodulators, such as immune checkpoint inhibitors and / or anticancer agents, such as anticancer agents that directly target tumor or cancer cells, has a low response to the one or more prior therapies, has not achieved a desired degree of response to the one or more prior therapies, has achieved less than a desired degree of response to the one or more prior therapies (e.g., has a poor response to the one or more prior therapies or is ineffective in treating the one or more prior therapies), or is unresponsive to the one or more prior therapies. In some embodiments, the tumor or cancer has achieved less than a desired degree of response or is predicted to be resistant to anti-PD-L1, anti-PD-1, and / or anti-CTLA-4 therapy. In some embodiments, the tumor or cancer achieves a less than desirable response or is predicted to be resistant to anti-PD-L1 therapy. In some embodiments, the tumor or cancer achieves a less than desirable response or is predicted to be resistant to anti-PD-1 therapy. In some embodiments, the tumor or cancer achieves a less than desirable response or is predicted to be resistant to anti-CTLA-4 therapy.

[0378] The cancer includes one or more primary tumors and metastatic tumor cells, such as metastatic cancer; newly arising tumors or cancers; cancers including one or more primary tumors; and / or invasive tumor cells, such as invasive cancer. In some aspects, the provided compositions, methods, uses, and combinations can also sensitize cold tumors, including primary cold tumors and secondary cold tumors (e.g., metastatic tumors), to immunomodulatory agents or other anti-cancer therapies.

[0379] Such methods and uses include, for example, administering an anti-PD-L1 conjugate to an individual with a tumor or tumor cell, followed by irradiation of the target area (e.g., a site where cells expressing PD-L1 are present) with a wavelength and dose of light suitable for the phthalocyanine dye. In some aspects, irradiation causes irradiation-dependent dissolution and death of cells expressing target molecules (e.g., PD-L1) on the surface, thereby producing a therapeutic effect or treatment of cancer. In some cases, cells expressing PD-L1, such as monocytes, macrophages, dendritic cells (DC), M2 tumor-associated macrophages (M2 TAM), tolerant dendritic cells (tDC), or myeloid-derived suppressor cells (MDSC), or certain tumor cells are killed and thereby rapidly exhausted. Therefore, necrosis of tumor cells may occur.

[0380] In some aspects, the tumors, lesions, or cancers treated according to the provided embodiments include those that have failed one or more prior therapies, such as immunomodulatory agents, e.g., immune checkpoint inhibitors and / or anti-cancer agents, such as anti-PD-L1, anti-PD-1, or anti-CTLA-4 therapy, are poorly responsive or substantially unresponsive to the one or more prior therapies, are less responsive to the one or more prior therapies, did not achieve a desired degree of response to the one or more prior therapies, achieved less than a desired degree of response to the one or more prior therapies (e.g., were poorly responsive to the one or more prior therapies or were ineffectively treated with the one or more prior therapies), relapsed after the one or more prior therapies, are refractory to the one or more prior therapies, and / or are resistant to the one or more prior therapies.

[0381] In some embodiments, the individual treated according to the provided embodiments has been previously treated with an anti-cancer therapy and / or an immune checkpoint inhibitor. In some embodiments, the individual treated according to the provided embodiments has been previously treated with an immune checkpoint inhibitor. In some embodiments, the individual treated according to the provided embodiments has failed a previous treatment with an anti-cancer therapy and / or an immune checkpoint inhibitor, or has relapsed after such a previous treatment. In some embodiments, the individual treated according to the provided embodiments has failed a previous treatment with an immune checkpoint inhibitor, or has relapsed after such a previous treatment.

[0382] In some embodiments, the inhibition of tumor growth resulting from performing the method is greater than the inhibition of tumor growth resulting from prior treatment with an anti-cancer therapy and / or an immune checkpoint inhibitor (e.g., anti-PD-L1, anti-PD-1, and / or anti-CTLA-4 therapy). In some embodiments, the inhibition of tumor growth resulting from performing the method is greater than the inhibition of tumor growth resulting from prior treatment with an immune checkpoint inhibitor (e.g., anti-PD-L1, anti-PD-1, and / or anti-CTLA-4 therapy).

[0383] In some aspects, the one or more prior therapeutic treatments to which the cancer is unresponsive include the use of an anticancer agent. The prior anticancer agent may be one or more of the following: a chemotherapeutic agent, an antibody therapy, and / or a radiotherapy agent. In some embodiments, the prior therapy is therapy with an anticancer agent selected from the group consisting of a checkpoint inhibitor, an immunoadjuvant, a chemotherapeutic agent, radiation, and a biologic comprising an anticancer targeting molecule that binds to tumor cells. In some embodiments, the prior therapy is therapy with an anticancer agent that is an antibody conjugate. In some embodiments, the prior therapy is therapy with an antibody conjugate comprising a phthalocyanine dye, a toxin, or a TLR agonist.

[0384] In some aspects, one or more prior therapeutic treatments to which the cancer, tumor, or tumor cells have not responded can be treatment with an immune checkpoint inhibitor (also known as immune checkpoint blockade therapy). The prior immune checkpoint inhibitor can be a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, or a combination thereof. The prior immune checkpoint inhibitor can be a small molecule inhibitor, an antibody inhibitor, or other molecules that bind to and inhibit immune checkpoint proteins, such as PD-1 or PD-L1. Exemplary antibody inhibitors for PD-1 include, but are not limited to, any of the following: pembrolizumab (MK-3475, Keytruda), nivolumab (OPDIVO), cemiplimab (LIBTAYO), toripalimab (JS001), HX008, SG001, GLS-010, dostarlimab (TSR-042), tislelizumab (BGB-A317), cetrel (Cetrel imab)(JNJ-63723283), pidilizumab (CT-011), genolimzumab (APL-501, GB226), BCD-100, siplizumab (REGN2810), F520, sintilimab (IBI308), GLS-010, CS1003, LZM009, camrelizumab (SHR-1210), SCT-I10A, MGA012, AK105, PF-06801591, AMP-224, AB 122, AMG 404, BI 754091, HLX10, JTX-4014, MEDI0680, Sym021, MGD019, MGD013, AK104, XmAb20717, RO7121661, CX-188, and spartalizumab.Exemplary antibody inhibitors for PD-L1 include, but are not limited to, any of the following: atezolizumab (MPDL3280A, Tecentriq), avelumab (Bavencio), durvalumab (MEDI4736, Imfinzi), LDP, NM-01, STI-3031, KN035, LY3300054, M7824 (MSB0011359C), BMS- 936559, MSB2311, BCD-135, BGB-A333, CBT-502, cosibellimab (CK-301), CS1001, FAZ053, MDX-1105, SHR-1316, TG-1501, ZKAB001, INBRX-105, MCLA-145, KN046, LY3415244, REGN3504, and HLX20.

[0385] In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that is resistant to, difficult to treat, or unresponsive to treatment with an anti-PD-1 antibody or an anti-PD-L1 antibody. In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that is resistant to, difficult to treat, or unresponsive to treatment with an anti-PD-L1 antibody, or is predicted to be unresponsive to, resistant to, or unresponsive to treatment with an anti-PD-L1 antibody. In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that is resistant to, difficult to treat, or unresponsive to treatment with an anti-PD-1 antibody, or is predicted to be unresponsive to, resistant to, or unresponsive to treatment with an anti-PD-1 antibody.

[0386] In some aspects, the prior treatment is treatment with an anti-CTLA-4 antibody, such as ipilimumab (YERVOY), tremelimumab, AGEN 1181, AGEN 1884, ADU-1064, BCD-145, and BCD-217. In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that is resistant, refractory, or unresponsive to anti-CTLA-4 antibody treatment, or is predicted to be unresponsive, resistant, or refractory to anti-CTLA-4 antibody treatment.

[0387] In some aspects, the one or more prior therapeutic treatments to which the cancer, tumor, or tumor cells have not responded can be an immunomodulator, such as a cytokine, for example, aldesleukin (PROLEUKIN), interferon alpha-2a, interferon alpha-2b (Intron A), pegylated interferon alpha-2b (SYLATRON / PEG-Intron), or a cytokine targeting the IFNAR1 / 2 pathway, the IL-2 / IL-2R pathway, or an adjuvant, for example, polyICLC (HILTONOL / Imiquimod), 4-1BB (CD137; TNFRS9), OX40 (CD134), OX40-ligand (OX40L), Toll-Like Receptors (TLRs), or a combination thereof. Therapies targeting the Toll-like receptor 7 (TLR7) pathway, TNFR and other members of the TNF superfamily, other TLR2 agonists, TLR3 agonists and TLR4 agonists are also included.

[0388] In some aspects, the cancer is unresponsive to one or more prior therapeutic treatments that include the use of a therapeutic agent that targets immunosuppressive cells. The agent can be an antibody that targets regulatory T cells, for example, an anti-CD25 antibody such as basiliximab. Daclizumab or PC61; small molecule inhibitors; or a combination thereof. Immunosuppressive cells include regulatory T cells, M2 macrophages, tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs), or a combination thereof.

[0389] In some cases, the tumors, lesions, or cancers treated according to the provided embodiments include "cold tumors" or "cold cancers," such as tumors with an immunosuppressive phenotype. Such cold tumors may have a variety of characteristics, including but not limited to intratumoral CD8 + A significant decrease or absence of the number and / or activity of T effector cells, and / or a significant increase in the number and / or activity of immunosuppressive cells within the tumor. In some cases, a cold tumor or cancer has a high tumor mutation burden (TMB), an immune score indicative of low immunoreactivity, or a programmed cell death protein 1 (PD-1) or programmed death-ligand 1 (PD-L1) marker status (e.g., cell surface expression) that can indicate low immunoreactivity. In some cases, a cold tumor or cancer does not respond to PD-1 or PD-L1 inhibitor monotherapy.

[0390] In some embodiments, cold tumors or cancers can be treated with an anti-PD-L1 conjugate as described herein, followed by irradiation. In some embodiments, combination treatment with an anti-PD-L1 conjugate followed by irradiation with an immunomodulatory agent such as an immune checkpoint inhibitor results in enhanced inhibition of growth of both the irradiated primary tumor and distant tumors.

[0391] In addition, for tumors that are resistant to treatment with immunomodulatory therapies, such as treatment with immune checkpoint inhibitors, treatment with anti-PD-L1 conjugates followed by photoirradiation and / or in combination with immune checkpoint inhibitors can enhance the inhibitory effect on the growth of irradiated primary tumors and distant tumors, primary tumors and newly arising tumors, and / or primary tumors and different types of secondary tumors, indicating that anti-PD-L1 photoimmunotherapy has a sensitizing effect on immune checkpoint inhibitors in the treatment of cancer and tumor cells.

[0392] IV. Administration Methods and Formulations

[0393] Compositions comprising PD-L1 antibodies, antigen-binding fragments, and immunoconjugates, including pharmaceutical compositions and formulations, are also provided.

[0394] In some embodiments, the anti-PD-L1 conjugate can be administered systemically or locally to the organ or tissue to be treated. Exemplary routes of administration include, but are not limited to, topical, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intratumoral, and intravenous), oral, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation routes. In some embodiments, the anti-PD-L1 conjugate is administered intravenously. In some embodiments, the anti-PD-L1 conjugate is administered parenterally. In some embodiments, the anti-PD-L1 conjugate is administered enterally. In some embodiments, the conjugate is administered via local injection. In some embodiments, the conjugate is administered in a topical administration form.

[0395] Compositions comprising anti-PD-L1 conjugates can be administered locally or systemically using any method known in the art, for example, to individuals with tumors, such as cancer, or individuals from whom tumors have previously been removed, for example, via surgery. Although specific examples are provided, those skilled in the art will appreciate that alternative methods of administration of the disclosed agents may be used. Such methods may include, for example, the use of a catheter or implantable pump to provide continuous infusion to an individual in need of treatment over a period of hours to days.

[0396] In some embodiments, the anti-PD-L1 conjugate is administered parenterally, including by direct injection or infusion into the tumor, such as intratumoral administration. In some embodiments, the anti-PD-L1 conjugate is administered to the tumor by applying the agent to the tumor, for example, by immersing the tumor in a solution containing the anti-PD-L1 conjugate or by pouring the agent onto the tumor.

[0397] Additionally or alternatively, the anti-PD-L1 conjugate can be administered systemically, e.g., intravenously, intramuscularly, subcutaneously, intradermally, intraperitoneally, subcutaneously, or orally, to an individual having a tumor, such as cancer.

[0398] Also provided herein are compositions containing anti-PD-L1 conjugates, such as pharmaceutical compositions, and uses of such compositions, such as therapeutic uses and / or uses as medicaments. In some aspects, the composition comprises an anti-PD-L1 conjugate and a pharmaceutically acceptable carrier. In some embodiments, the composition containing the anti-PD-L1 conjugate is for use in a treatment or therapy according to any one of the embodiments provided, such as for administration to an individual suffering from a disease or condition, for treating the disease or condition. There is no absolute limit to the dose of the anti-PD-L1 conjugate to be administered to an individual, but will depend on the properties of the composition and its active ingredients and its unwanted side effects, such as the immune response to the agent, the individual being treated, and the type and mode of administration of the condition being treated. In general, the dose will be a therapeutically effective amount, such as an amount sufficient to achieve the desired biological effect, for example, an amount that is effective to reduce the size of the tumor (such as volume and / or weight) or to alleviate further growth of the tumor or to reduce the undesirable symptoms of the tumor.

[0399] In some embodiments, compositions for administering an anti-PD-L1 conjugate contain an effective amount of the agent and conventional pharmaceutical carriers and excipients appropriate for the type of administration contemplated. For example, in some embodiments, parenteral formulations may contain a sterile aqueous solution or suspension of the conjugate. In some embodiments, compositions for enteral administration may contain an effective amount of an anti-PD-L1 conjugate in an aqueous solution or suspension, which may optionally include a buffer, a surfactant, a thixotropic agent, and a flavoring agent.

[0400] In some embodiments, the anti-PD-L1 conjugate or the combination of the conjugate and the additional therapeutic agent can be formulated in a pharmaceutically acceptable buffer, such as a pharmaceutically acceptable buffer containing a pharmaceutically acceptable carrier or vehicle. In general, pharmaceutically acceptable carriers or vehicles, such as those present in a pharmaceutically acceptable buffer, can be any known in the art. Remington's Pharmaceutical Sciences, EW Martin, Mack Publishing Co., Easton, Pennsylvania, 19th edition (1995) describes compositions and formulations suitable for drug delivery of one or more therapeutic compounds. Pharmaceutically acceptable compositions are prepared according to generally recognized pharmacopeias for use in animals and humans, subject to approval by regulatory agencies or other agencies.

[0401] The pharmaceutical composition may include a carrier, such as a diluent, adjuvant, excipient, or vehicle, to be administered with the compound. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. Such compositions will contain a therapeutically effective amount of the compound, generally in purified form, and a suitable amount of carrier to provide a form suitable for administration to the patient. Such pharmaceutical carriers may be sterile liquids, such as water and oils, including oils of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. When the pharmaceutical composition is administered intravenously, water is a typical carrier. Physiological saline solutions and aqueous dextrose and glycerol solutions may also be used as liquid carriers, particularly for injectable solutions. In addition to the active ingredient, the composition may also contain: diluents such as lactose, sucrose, dicalcium phosphate, or carboxymethylcellulose; lubricants such as magnesium stearate, calcium stearate, and talc; and binders such as starch, natural gums such as acacia gelatin, glucose, molasses, polyvinylpyrrolidone, cellulose and its derivatives, povidone, crospovidone, and other such binders known to those skilled in the art. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, ethylene glycol, water, and ethanol. If necessary, the composition may also contain small amounts of wetting agents or emulsifiers, or pH buffers, such as acetate, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, sodium triethanolamine acetate, triethanolamine oleate, and other such agents.

[0402] In some embodiments, the pharmaceutical preparation may be in liquid form, for example, in the form of a solution, syrup, or suspension. Such liquid preparations can be prepared in a conventional manner with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats); emulsifiers (e.g., lecithin or gum arabic); non-aqueous carriers (e.g., almond oil, oily esters, ethanol, or fractionated vegetable oils); and preservatives (e.g., methylparaben or propylparaben or sorbic acid). In some cases, the pharmaceutical preparation may be in a lyophilized form reconstituted with water or other suitable carriers prior to use.

[0403] In some embodiments, the nature of the pharmaceutically acceptable buffer or carrier depends on the specific mode of administration used. For example, in some embodiments, parenteral formulations can include injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, saline, balanced salt solutions, aqueous dextrose, or glycerol as a carrier. In some embodiments, for solid compositions, such as powders, pills, tablets, or capsules, non-toxic solid carriers can include, for example, pharmaceutical grade mannitol, lactose, starch, or magnesium stearate. Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed, and include, but are not limited to, buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexahydroxyquaternium chloride; benzalkonium chloride; benzethonium chloride; phenolic, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) ) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as polyethylene glycol (PEG). In some embodiments, in addition to the biologically neutral carrier, the pharmaceutical composition to be administered may contain minor amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, preservatives, and pH buffers, for example, sodium acetate or sorbitan monolaurate.

[0404] In some aspects, a buffer is included in the composition. Suitable buffers include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffers is used. The amount of the buffer or its mixture generally accounts for about 0.001% by weight to about 4% by weight of the total composition. Methods for preparing pharmaceutical compositions that can be administered are known. Exemplary methods are described in more detail in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st edition (May 1, 2005).

[0405] Formulations of the antibodies described herein may include lyophilized formulations and aqueous solutions.

[0406] The preparation or composition may also contain more than one active ingredient suitable for a specific indication, disease or condition treated with an antibody, antigen-binding fragment or conjugate, preferably those active ingredients having an activity complementary to the antibody, antigen-binding fragment or conjugate, wherein each activity has no adverse effect on each other. Such active ingredients are preferably present in an amount combination that effectively achieves the intended purpose. Therefore, in some embodiments, the pharmaceutical composition further includes other pharmaceutically active agents or drugs, such as chemotherapeutic agents, for example, asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, cranberries, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0407] The compound can be formulated into a suitable pharmaceutical preparation for oral administration, such as a solution, suspension, tablet, dispersible tablet, pill, capsule, powder, sustained release formulation or elixir, as well as a transdermal patch formulation and a dry powder inhaler. Typically, the compound is formulated into a pharmaceutical composition using techniques and procedures well known in the art (see, for example, Ansel, "Introduction to Pharmaceutical Dosage Forms" (Ansel Introduction to Pharmaceutical Dosage Forms), 4th edition, 1985, 126). In general, the mode of preparation is a function of the route of administration.

[0408] In some aspects, the pharmaceutical composition can be delivered using a timed-release, delayed-release, or sustained-release delivery system so that the composition is delivered before the site to be treated becomes sensitized and sufficient time has elapsed for the site to become sensitized. Various types of release delivery systems are available and known. Such systems can avoid repeated administration of the composition, thereby increasing convenience for both the individual and the physician.

[0409] In some embodiments, the pharmaceutical composition contains an amount that effectively treats or prevents a disease or condition, such as an antibody, antigen-binding fragment, or conjugate that is effective in treating or preventing an effective amount. In some embodiments, the therapeutic or preventive efficacy is monitored by regularly evaluating the individual being treated. For repeated administration over several days or longer, depending on the condition, repeated treatment is performed until the desired suppression of disease symptoms occurs. However, other dosing regimens are applicable and can be determined. The desired dose can be delivered via a single bolus, via multiple boluses, or via continuous infusion.

[0410] The composition can be formulated to be administered via any route known to those skilled in the art, including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, surface, topical, otic, inhalation, buccal (e.g., sublingual) and transdermal administration or any route. Other modes of administration are also contemplated. Administration can be local, topical, or systemic, depending on the treatment location. Topical administration to an area requiring treatment can be achieved, for example, but not limited to, local infusion during surgery, topical application (e.g., postoperatively with a wound dressing), via injection, with the aid of a catheter, by means of a suppository, or by means of an implant.

[0411] Parenteral administration is contemplated herein and is generally characterized by subcutaneous, intramuscular, intratumoral, intravenous, or intradermal injection. Injectables may be in conventional form, in the form of liquid solutions or suspensions, in solid forms suitable for dissolution or suspension prior to injection, or prepared in the form of emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if necessary, the pharmaceutical composition to be administered may also contain an activator in the form of a solvent, such as a pH buffer, a metal ion salt, or other such buffer. The pharmaceutical composition may also contain other small amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, stabilizers, solubility enhancers, and other such agents, such as sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins. Implantation of slow-release or sustained-release systems is also contemplated herein to maintain a constant dosage level (see, for example, U.S. Patent No. 3,710,795). The percentage of active compound contained in such parenteral compositions depends primarily on its specific properties, as well as the activity of the compound and individual needs.

[0412] Injectables are designed for local and systemic administration. Preparations for parenteral administration include sterile solutions that can be used immediately for injection, sterile anhydrous soluble products (such as lyophilized powders) (including subcutaneous lozenges) that can be combined with solvents immediately before use, sterile suspensions that can be used immediately for injection, sterile anhydrous insoluble products and sterile emulsions that can be combined with carriers immediately before use. The solution can be an aqueous solution or a non-aqueous solution. If given intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening agents and solubilizing agents, such as glucose, polyethylene glycol and polypropylene glycol and mixtures thereof.

[0413] Pharmaceutically acceptable carriers for parenteral preparations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending agents and dispersants, emulsifiers, chelating agents or chelating agents, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose, and lactated Ringer's injection. Non-aqueous parenteral vehicles include fixed oils of plant origin, cottonseed oil, corn oil, sesame oil, and peanut oil. Antimicrobial agents at bacteriostatic or antifungal concentrations can be added to parenteral preparations packaged in multidose containers, including phenol or cresol, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl parahydroxybenzoates, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate.

[0414] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0415] The composition can be formulated for single-dose administration or for multiple-dose administration. The agent can be formulated for direct administration. The composition can be provided in liquid or lyophilized form. When the composition is provided in lyophilized form, it can be reconstituted with an appropriate buffer, such as sterile saline solution, before use.

[0416] The compositions can also be administered continuously, intermittently or in the same composition with other biologically active agents. Administration can also include controlled release systems, which include controlled release formulations and devices such as pumps to control release.

[0417] The most appropriate route in any given case depends on a variety of factors, such as the nature of the disease, the progression of the disease, the severity of the disease, and the specific composition used. For example, the composition is administered systemically, for example, via intravenous administration. Subcutaneous administration can also be used, although it may require an increased absorption time compared to intravenous administration to ensure comparable bioavailability.

[0418] Pharmaceutical compositions can be formulated into dosage forms suitable for various routes of administration. Drugs and therapeutically active compounds and their derivatives are typically formulated and administered in unit dosage forms or multiple dosage forms. Each unit dose contains a predetermined amount of the therapeutically active compound sufficient to produce the desired therapeutic effect, in combination with the desired pharmaceutical carrier, vehicle, or diluent. Unit dosage forms include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, as well as oil-in-water emulsions containing an appropriate amount of the compound or its pharmaceutically acceptable derivatives. Unit dosage forms may contain ampoules and syringes or individually packaged tablets or capsules. Unit dosage forms can be administered in fractional or multiple portions. A multidose form is a plurality of identical unit dosage forms packaged in a single container to be administered in separate unit dosage forms. Examples of multidose forms include vials, bottles of tablets or capsules, or pint or gallon bottles. Thus, a multidose form is a plurality of unit doses that are not separated in the packaging. Generally speaking, dosage forms or compositions containing active ingredients in the range of 0.005% to 100% can be prepared with the remainder consisting of non-toxic carriers. Pharmaceutical compositions can be formulated into dosage forms suitable for each route of administration.

[0419] The concentration of the pharmaceutically active compound is adjusted so that the injection provides an amount that is effective in producing the desired pharmacological effect. As is known in the art, the exact dosage depends on the age, weight, and condition of the patient or animal. Unit dose parenteral formulations are packaged in ampoules, vials, or syringes with needles. The volume of the liquid solution or reconstituted powder formulation containing the pharmaceutically active compound varies with the disease to be treated and the specific product selected for packaging. As is known and practiced in the art, all formulations for parenteral administration must be sterile. In some embodiments, the composition can be provided in the form of a lyophilized powder, which can be reconstituted to be administered in the form of a solution, emulsion, or other mixture. It can also be reconstituted and formulated into a solid or gel form. The lyophilized powder can be prepared from any of the above solutions.

[0420] Sterile lyophilized powders can be prepared by dissolving the phthalocyanine dye-targeting molecule conjugate in a buffered solution. The buffered solution may contain excipients that improve the stability of the powder or a reconstituted solution prepared from the powder or other pharmacological components.

[0421] In some embodiments, the solution is then sterile filtered and then lyophilized under standard conditions known to those skilled in the art to obtain the desired formulation. In short, lyophilized powder is prepared by dissolving excipients such as dextrose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose or other suitable reagents in a suitable buffer, such as citrate, sodium phosphate or potassium phosphate or other such buffers known to those skilled in the art. Subsequently, the selected enzyme is added to the resulting mixture and stirred until it dissolves. The resulting mixture is sterile filtered or treated to remove particles and ensure sterility, and is distributed into vials for lyophilization. Each vial can contain a single dose (1 mg-1 g, generally 1-100 mg, such as 1-5 mg) or multiple doses of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4 ° C to room temperature. Reconstructing this lyophilized powder with an appropriate buffer solution gives a preparation for parenteral administration. The exact amount depends on the indication being treated and the selected compound. This amount can be determined empirically.

[0422] In some embodiments, the pH of the composition is between 6 and 10 or about 6 and 10, such as between 6 and 8 or about 6 and 8, between 6.9 and 7.3 or about 6.9 and 7.3, such as about pH 7.1. In some embodiments, the pH of the pharmaceutically acceptable buffer is at least 5 or about 5, at least 6 or about 6, at least 7 or about 7, at least 8 or about 8, at least 9 or about 9, or at least 10 or about 10, or 7.1.

[0423] The composition can be formulated for single dose administration or for multiple dose a...

Claims

1. An antibody or antigen-binding fragment that specifically binds to a programmed death ligand (PD-L1) protein, wherein the antibody or antigen-binding fragment comprises: (a) a heavy chain variable (VH) region, the VH region comprising a heavy chain complementary determining region 1 (CDR-H1), CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO: 1, respectively; and a light chain variable (VL) region, the VL region comprising a light chain complementary determining region 1 (CDR-L1), CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO: 17, respectively; (b) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:2, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:18, respectively; (c) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:3; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:19; (d) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:4, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:19, respectively; (e) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise the CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:5, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise the CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:20, respectively; (f) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO: 1, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO: 21, respectively; (g) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:2, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:22, respectively; (h) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise the CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:6; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise the CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:23; (i) a VH region, wherein the VH region comprises CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence set forth in SEQ ID NO:7, respectively; and a VL region, wherein the VL region comprises CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence set forth in SEQ ID NO:24, respectively; (j) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:8, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:25, respectively; (k) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:9, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:26, respectively; (1) a VH region, wherein the VH region comprises CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence set forth in SEQ ID NO: 10, respectively; and a VL region, wherein the VL region comprises CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence set forth in SEQ ID NO: 27, respectively; (m) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO: 11, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO: 28, respectively; (n) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence set forth in SEQ ID NO: 12, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence set forth in SEQ ID NO: 29, respectively; (o) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise the CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO: 13, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise the CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO: 30, respectively; (p) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, comprising CDR-H1, CDR-H2 and CDR-H3, respectively, contained in the VH region amino acid sequence set forth in SEQ ID NO: 14; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, comprising CDR-L1, CDR-L2 and CDR-L3, respectively, contained in the VL region amino acid sequence set forth in SEQ ID NO: 31; (q) a VH region, the VH region comprising CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise the CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO: 15, respectively; and a VL region, the VL region comprising CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise the CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO: 32, respectively; (r) a VH region, wherein the VH region comprises CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3, respectively, contained in the VH region amino acid sequence set forth in SEQ ID NO: 12; and a VL region, wherein the VL region comprises CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3, respectively, contained in the VL region amino acid sequence set forth in SEQ ID NO: 33; or (s) a VH region, wherein the VH region comprises CDR-H1, CDR-H2 and CDR-H3, respectively, which comprise CDR-H1, CDR-H2 and CDR-H3 contained in the VH region amino acid sequence shown in SEQ ID NO:16, respectively; and a VL region, wherein the VL region comprises CDR-L1, CDR-L2 and CDR-L3, respectively, which comprise CDR-L1, CDR-L2 and CDR-L3 contained in the VL region amino acid sequence shown in SEQ ID NO:34, respectively.

2. The antibody or antigen-binding fragment of claim 1, wherein: (a) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:40, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:41, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:37; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:210, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:211, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:212; (b) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:52, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:53, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:37; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:218, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:211, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:212; (c) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:63, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:64, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:60; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:221, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:222, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:223; (d) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:63, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:64, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:60; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:221, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:222, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:223; (e) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 75, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 76, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 60; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 229, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 222, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 223; (f) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:40, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:41, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:37; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:233, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:234, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:235; (g) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:52, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:53, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:37; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:241, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:234, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:242; (h) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 87, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 88, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 84; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 246, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 247, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 248; (i) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO:98, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:99, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:84; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO:246, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:254, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:255; (j) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 108, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 109, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 105; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 258, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 259, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 260; (k) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 121, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 122, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 118; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 265, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 266, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 267; (1) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 134, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 135, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 131; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 246, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 273, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 274; (m) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 147, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 148, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 144; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 278, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 279, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 280; (n) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 160, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 161, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 157; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 286, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 287, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 288; (o) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 173, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 174, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 170; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 294, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 234, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 295; (p) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 185, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 186, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 182; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 299, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 300, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 301; (q) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 173, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 174, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 193; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 306, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 234, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 307; (r) the VH region comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 160, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 161, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 157; and the VL region comprises: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 311, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 312, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 313; or (s) the VH region comprises: CDR-H1 comprising the amino acid sequence shown in SEQ ID NO:202, CDR-H2 comprising the amino acid sequence shown in SEQ ID NO:203, and CDR-H3 comprising the amino acid sequence shown in SEQ ID NO:199; and the VL region comprises: CDR-L1 comprising the amino acid sequence shown in SEQ ID NO:319, CDR-L2 comprising the amino acid sequence shown in SEQ ID NO:320, and CDR-L3 comprising the amino acid sequence shown in SEQ ID NO:

321.

3. The antibody or antigen-binding fragment of claim 1 or 2, wherein: (a) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 1; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 17; (b) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 2; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 18; (c) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:3; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:19; (d) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:4; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:19; (e) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:5; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:20; (f) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 1; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 21; (g) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:2; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:22; (h) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:6; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:23; (i) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:7; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:24; (j) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:8; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:25; (k) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:9; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:26; (l) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:10; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:27; (m) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 11; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 28; (n) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 12; and the VL region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 29; (o) the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 13; and the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 30; (p) the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 14; and the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 31; (q) the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 15; and the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 32; (r) the VH region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 12; and the VL region comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 33; or (s) the VH region comprises an amino acid sequence that has at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO:16; and the VL region comprises an amino acid sequence that has at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO:

34.

4. The antibody or antigen-binding fragment of any one of claims 1 to 3, wherein: (a) the VH region comprises the amino acid sequence shown in SEQ ID NO: 1, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 17; (b) the VH region comprises the amino acid sequence shown in SEQ ID NO: 2, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 18; (c) the VH region comprises the amino acid sequence shown in SEQ ID NO:3, and the VL region comprises the amino acid sequence shown in SEQ ID NO:19; (d) the VH region comprises the amino acid sequence shown in SEQ ID NO:4, and the VL region comprises the amino acid sequence shown in SEQ ID NO:19; (e) the VH region comprises the amino acid sequence shown in SEQ ID NO:5, and the VL region comprises the amino acid sequence shown in SEQ ID NO:20; (f) the VH region comprises the amino acid sequence shown in SEQ ID NO: 1, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 21; (g) the VH region comprises the amino acid sequence shown in SEQ ID NO: 2, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 22; (h) the VH region comprises the amino acid sequence shown in SEQ ID NO:6, and the VL region comprises the amino acid sequence shown in SEQ ID NO:23; (i) the VH region comprises the amino acid sequence shown in SEQ ID NO:7, and the VL region comprises the amino acid sequence shown in SEQ ID NO:24; (j) the VH region comprises the amino acid sequence shown in SEQ ID NO:8, and the VL region comprises the amino acid sequence shown in SEQ ID NO:25; (k) the VH region comprises the amino acid sequence shown in SEQ ID NO:9, and the VL region comprises the amino acid sequence shown in SEQ ID NO:26; (1) the VH region comprises the amino acid sequence shown in SEQ ID NO: 10, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 27; (m) the VH region comprises the amino acid sequence shown in SEQ ID NO: 11, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 28; (n) the VH region comprises the amino acid sequence shown in SEQ ID NO: 12, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 29; (o) the VH region comprises the amino acid sequence shown in SEQ ID NO: 13, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 30; (p) the VH region comprises the amino acid sequence shown in SEQ ID NO: 14, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 31; (q) the VH region comprises the amino acid sequence shown in SEQ ID NO: 15, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 32; (r) the VH region comprises the amino acid sequence shown in SEQ ID NO: 12, and the VL region comprises the amino acid sequence shown in SEQ ID NO: 33; or (s) the VH region comprises the amino acid sequence shown in SEQ ID NO:16, and the VL region comprises the amino acid sequence shown in SEQ ID NO:

34.

5. The antibody or antigen-binding fragment of any one of claims 1 to 4, wherein the PD-L1 protein is a human PD-L1 protein.

6. The antibody or antigen-binding fragment of any one of claims 1 to 5, wherein the antibody or antigen-binding fragment is recombinant.

7. The antibody or antigen-binding fragment of any one of claims 1 to 6, wherein the antibody or antigen-binding fragment is monoclonal.

8. The antibody or antigen-binding fragment of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment is a human, chimeric or humanized antibody or antigen-binding fragment.

9. The antibody or antigen-binding fragment of any one of claims 1 to 8, wherein the antibody or antigen-binding fragment comprises an Fc region of a human immunoglobulin and / or a human antibody framework region.

10. The antibody or antigen-binding fragment of any one of claims 1 to 9, which is a single-chain antibody fragment.

11. The antibody or antigen-binding fragment of claim 10, wherein the antibody fragment comprises a single-chain Fv (scFv).

12. The antibody or antigen-binding fragment of any one of claims 1 to 11, which is a complete or intact antibody.

13. The antibody or antigen-binding fragment of any one of claims 1 to 12, which is a bispecific antibody that further specifically binds to a second antigen.

14. The antibody or antigen-binding fragment of claim 13, wherein the second antigen is an antigen expressed on tumor cells or immune cells.

15. The antibody or antigen-binding fragment of claim 13 or 14, wherein the second antigen is CD25.

16. The antibody or antigen-binding fragment of any one of claims 1 to 15, wherein the antibody or antigen-binding fragment thereof comprises an Fc region that exhibits one or more Fc-mediated effector functions.

17. The antibody or antigen-binding fragment of any one of claims 1 to 15, wherein the antibody or antigen-binding fragment thereof comprises an Fc region that lacks Fc-mediated effector function, exhibits significantly reduced Fc-mediated effector function, or does not exhibit significant Fc-mediated effector function.

18. The antibody or antigen-binding fragment of any one of claims 1 to 15, wherein the antibody or antigen-binding fragment thereof comprises an Fc region that exhibits enhanced Fc-mediated effector functions.

19. The antibody or antigen-binding fragment of any one of claims 16 to 18, wherein the Fc-mediated effector function is selected from one or more of antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC).

20. The antibody or antigen-binding fragment of any one of claims 1 to 19, wherein the antibody or antigen-binding fragment comprises an IgG1 Fc region or IgG1 isotype, an IgG2 Fc region or IgG2 isotype, an IgG3 Fc region or IgG3 isotype, or an IgG4 Fc region or IgG4 isotype.

21. A conjugate comprising the antibody or antigen-binding fragment of any one of claims 1 to 20 and a heterologous molecule or moiety.

22. The conjugate of claim 21, wherein the heterologous molecule or moiety is a protein, a peptide, a nucleic acid, a dye or a small molecule.

23. The conjugate of claim 21 or 22, wherein the heterologous molecule or moiety is a cytotoxic agent, a toxin, a radioisotope, a chemotherapeutic agent, a lytic peptide, a cytokine, or a photoactivated dye.

24. The conjugate of claim 23, wherein the photoactivated dye is a phthalocyanine dye. The conjugate of claim 24 , wherein the phthalocyanine dye is a Si-phthalocyanine dye.

26. The conjugate of claim 24 or 25, wherein the phthalocyanine dye is IR700.

27. The conjugate of claim 24 or 25, wherein the phthalocyanine dye has the structure of formula (I): or a salt, stereoisomer or tautomer thereof.

28. The conjugate of any one of claims 21 to 27, wherein the conjugate is activated to achieve cell killing via irradiation at a wavelength between at or about 600 nm and at or about 850 nm.

29. The conjugate of claim 28, wherein the activated conjugate achieves tumor growth inhibition or killing at a higher level, activity or potency than the unconjugated antibody.

30. The conjugate of any one of claims 21 to 29, wherein the antibody or antigen binding fragment and the moiety are linked directly or indirectly via a linker.

31. The conjugate of any one of claims 21 to 29, wherein the antibody or antigen binding fragment is covalently linked to the heterologous molecule or moiety.

32. The conjugate of any one of claims 21 to 31, wherein when contacted with a cell expressing the PD-L1 protein, the conjugate exhibits increased internalization compared to an unconjugated antibody or antigen-binding fragment or a conjugate comprising a reference antibody.

33. The conjugate of any one of claims 21 to 31, wherein when contacted with a cell expressing the PD-L1 protein, the conjugate exhibits reduced internalization compared to an unconjugated antibody or antigen-binding fragment or a conjugate comprising a reference antibody.

34. The conjugate of claim 32 or 33, wherein the reference antibody is avelumab.

35. The conjugate of any one of claims 21 to 34, wherein the conjugate does not exhibit a significantly reduced binding affinity for the PD-L1 protein compared to the unconjugated antibody, or exhibits at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% of the binding affinity of the unconjugated antibody for the PD-L1 protein.

36. A polynucleotide encoding the antibody or antigen-binding fragment of any one of claims 1 to 20.

37. A vector comprising the polynucleotide of claim 36.

38. The vector of claim 37, wherein the vector is an expression vector.

39. An engineered cell comprising the vector of claim 37 or 38.

40. An engineered cell expressing the antibody or antigen-binding fragment of any one of claims 1 to 20.

41. A composition comprising the antibody or antigen-binding fragment thereof as described in any one of claims 1 to 20 or the conjugate as described in any one of claims 21 to 35.

42. The composition of claim 41, further comprising a pharmaceutically acceptable excipient.

43. A method of treatment comprising administering to an individual suffering from a disease or condition an antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 20, a conjugate as claimed in any one of claims 21 to 35, or a composition as claimed in claim 41 or 42.

44. The method of claim 43, wherein the disease or disorder is a tumor or cancer.

45. A method of treating a tumor or lesion in an individual, comprising: a) administering to the individual the conjugate of any one of claims 21 to 35 or the composition of claim 41 or 42; and b) using a wavelength between at or about 600 nm and at or about 850 nm and 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 or irradiating a target area in the individual at a dose of at or about 2 J / cm of fiber length to at or about 500 J / cm of fiber length, thereby activating the conjugate; Thereby the growth, volume or size of the tumor or lesion is reduced or inhibited.

46. ​​The method of any one of claims 43 to 45, wherein the method results in killing of cells expressing PD-L1 in the target area.

47. The method of any one of claims 43 to 46, wherein the individual has a tumor or lesion that is poorly responsive, unresponsive, resistant, refractory to, unresponsive to, or relapsed following a previous immunotherapy.

48. The method of claim 47, wherein the prior immunotherapy is treatment with an immune checkpoint inhibitor.

49. The method of claim 47 or 48, wherein the individual has primary or acquired resistance to a prior immunotherapy comprising PD-1:PD-L1 blockade therapy.

50. The method of any one of claims 43 to 49, wherein the individual is not treated with or has not previously received treatment with an immune checkpoint inhibitor.

51. The method of any one of claims 43 to 50, wherein the conjugate is administered to the subject to treat, inhibit growth and / or reduce size of a first tumor or lesion; and the method inhibits, delays or prevents the emergence, growth or establishment of one or more second tumors or lesions distal to the first tumor or lesion.

52. A method of immunizing an individual having a first tumor or lesion, the method comprising: (a) administering to an individual a conjugate according to any one of claims 21 to 35 or a composition according to claim 41 or 42; and (b) at a wavelength of between about 600 nm and about 850 nm and at a rate of 25 J / cm 2 or about 25J / cm 2 Up to 400J / cm 2 or about 400 J / cm 2 or irradiating a target area within the first tumor or lesion with a dose of at or about 2 J / cm of fiber length to at or about 500 J / cm of fiber length; wherein the growth of the first tumor or lesion is inhibited and / or the size is reduced; and the emergence, growth or establishment of one or more second tumors or lesions distal to the treated first tumor or lesion is inhibited, delayed or prevented.

53. The method of claim 51 or 52, wherein the second tumor or lesion is a metastasis of the first tumor or lesion.

54. The method of any one of claims 46 to 53, wherein the cell expressing PD-L1 is an immune cell.

55. The method of claim 54, wherein the immune cell is a monocyte, a macrophage, a dendritic cell (DC) or a myeloid-derived suppressor cell (MDSC).

56. The method of any one of claims 45 to 55, wherein the irradiating is performed between 30 minutes and 96 hours after administration of the conjugate or at or about 24 hours ± 4 hours after administration of the conjugate.

57. The method of any one of claims 45 to 56, wherein the target area is irradiated at a wavelength of 690±40 nm.

58. The method of any one of claims 45 to 57, wherein the target area is irradiated at a wavelength of 670±50 nm.

59. The method of any one of claims 45 to 58, wherein the target area is exposed to 50 J / cm 2 or about 50J / cm 2 Or irradiated at a dose of 100 J / cm fiber length or about 100 J / cm fiber length.

60. The method of any one of claims 44 to 59, wherein the tumor, lesion or cancer is associated with a cancer selected from the group consisting of colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell carcinoma, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, small intestine cancer, spindle cell neoplasms, liver cancer, hepatoma, peripheral nerve cancer, brain cancer, skeletal muscle cancer, smooth muscle cancer, bone cancer, adipose tissue cancer, cervical cancer, uterine cancer, genital cancer, lymphoma and multiple myeloma.

61. The method of any one of claims 43 to 60, wherein one or more steps of the method are repeated.

62. The method of any one of claims 43 to 61, wherein administration of the antibody or antigen-binding fragment, the conjugate or the composition is repeated one or more times, optionally wherein after each repeated administration of the conjugate or the composition, the irradiation step is repeated.

63. The method of any one of claims 43 to 62, further comprising administering an additional therapeutic agent or anti-cancer therapy.

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