Anti-Programmed Cell Death Ligand 1 (PD-L1) Antibody Molecules, Polynucleotides Encoding Same, and Methods of Use

Abstract

Anti-programmed cell death ligand (PD-L1) antibodies, antigen-binding fragments thereof, multispecific antibodies, and polynucleotides encoding them are provided herein. PD-L1-targeted phthalocyanine dye conjugates, compositions and products containing the conjugates, and methods for administering them to a subject for photoimmunotherapy are also provided. In some embodiments, the anti-PD-L1 antibodies, fragments, and conjugates specifically bind to human PD-L1. Among the anti-PD-L1 antibodies are human antibodies and humanized antibodies. Methods and uses utilizing the provided anti-PD-L1 antibodies, antigen-binding fragments, and conjugates, for example, for the treatment of human patients, are also provided. TIFF2025508739000050.tif145168

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Application US 63 / 311,924, filed 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.

[0002] INCORPORATION BY REFERENCE TO SEQUENCE LISTING 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.

[0003] Field The present disclosure relates to programmed cell death ligand (PD-L1) binding molecules, particularly anti-PD-L1 antibodies, e.g., 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 technology]

[0004] background Programmed cell death ligand 1 (PD-L1), also known as cluster of differentiation 247 (CD247) or B7-H1, is a protein receptor that functions as an immune checkpoint and downregulates immune responses. Therapies involving inhibitors of the PD-1:PD-L1 pathway have been used in the treatment of cancer, but challenges remain. There remains an urgent need for improved compositions and methods to address clinical challenges. Embodiments that meet such needs are provided. Summary of the Invention

[0005] overview Provided herein are antibodies or antigen-binding fragments that specifically bind to a programmed cell death-ligand (PD-L1) protein. In some of any of the embodiments, the PD-L1-binding antibody or antigen-binding fragment comprises a heavy chain variable (VH) region comprising CDR-H1, CDR-H2, and CDR-H3, each of which comprises CDR-H1, CDR-H2, and CDR-H3, each of which comprises CDR-H1, CDR-H2, and CDR-H3, each of which comprises CDR-H1, CDR-H2, and CDR-H3, each of which comprises CDR-H1, CDR-L2, and CDR-L3, each of which comprises CDR-L ... 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise 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, which comprise 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, which comprise the 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, which comprise the 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, which comprise 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, which comprise 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise 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, which comprise 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the 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, which comprise the CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:34.

[0006] In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:35, CDR-H2 comprising SEQ ID NO:36, and CDR-H3 comprising SEQ ID NO:37; the VL region comprises CDR-L1 comprising SEQ ID NO:210, CDR-L2 comprising SEQ ID NO:211, and CDR-L3 comprising SEQ ID NO:212. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:48, CDR-H2 comprising SEQ ID NO:49, and CDR-H3 comprising SEQ ID NO:37; and the VL region comprises CDR-L1 comprising SEQ ID NO:218, CDR-L2 comprising SEQ ID NO:211, and CDR-L3 comprising SEQ ID NO:212. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:58, CDR-H2 comprising SEQ ID NO:59, and CDR-H3 comprising SEQ ID NO:60; the VL region comprises CDR-L1 comprising SEQ ID NO:221, CDR-L2 comprising SEQ ID NO:222, and CDR-L3 comprising SEQ ID NO:223. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:71, CDR-H2 comprising SEQ ID NO:72, and CDR-H3 comprising SEQ ID NO:60; the VL region comprises CDR-L1 comprising SEQ ID NO:229, CDR-L2 comprising SEQ ID NO:222, and CDR-L3 comprising SEQ ID NO:223. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:48, CDR-H2 comprising SEQ ID NO:49, and CDR-H3 comprising SEQ ID NO:37; the VL region comprises CDR-L1 comprising SEQ ID NO:241, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:242. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:82, CDR-H2 comprising SEQ ID NO:83, and CDR-H3 comprising SEQ ID NO:84; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:247, and CDR-L3 comprising SEQ ID NO:248. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:48, CDR-H2 comprising SEQ ID NO:95, and CDR-H3 comprising SEQ ID NO:84; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:254, and CDR-L3 comprising SEQ ID NO:255. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:48, CDR-H2 comprising SEQ ID NO:104, and CDR-H3 comprising SEQ ID NO:105; the VL region comprises CDR-L1 comprising SEQ ID NO:258, CDR-L2 comprising SEQ ID NO:259, and CDR-L3 comprising SEQ ID NO:260. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:129, CDR-H2 comprising SEQ ID NO:130, and CDR-H3 comprising SEQ ID NO:131; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:273, and CDR-L3 comprising SEQ ID NO:274. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:142, CDR-H2 comprising SEQ ID NO:143, and CDR-H3 comprising SEQ ID NO:144; the VL region comprises CDR-L1 comprising SEQ ID NO:278, CDR-L2 comprising SEQ ID NO:279, and CDR-L3 comprising SEQ ID NO:280. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:155, CDR-H2 comprising SEQ ID NO:156, and CDR-H3 comprising SEQ ID NO:157; the VL region comprises CDR-L1 comprising SEQ ID NO:286, CDR-L2 comprising SEQ ID NO:287, and CDR-L3 comprising SEQ ID NO:288. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:168, CDR-H2 comprising SEQ ID NO:169, and CDR-H3 comprising SEQ ID NO:170; the VL region comprises CDR-L1 comprising SEQ ID NO:294, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:295. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:168, CDR-H2 comprising SEQ ID NO:169, and CDR-H3 comprising SEQ ID NO:193; the VL region comprises CDR-L1 comprising SEQ ID NO:306, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:307. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:155, CDR-H2 comprising SEQ ID NO:156, and CDR-H3 comprising SEQ ID NO:157; the VL region comprises CDR-L1 comprising SEQ ID NO:311, CDR-L2 comprising SEQ ID NO:312, and CDR-L3 comprising SEQ ID NO:313. In some of any of the 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.

[0007] In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:40, CDR-H2 comprising SEQ ID NO:41, and CDR-H3 comprising SEQ ID NO:37; the VL region comprises CDR-L1 comprising SEQ ID NO:210, CDR-L2 comprising SEQ ID NO:211, and CDR-L3 comprising SEQ ID NO:212. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:52, CDR-H2 comprising SEQ ID NO:53, and CDR-H3 comprising SEQ ID NO:37; the VL region comprises CDR-L1 comprising SEQ ID NO:218, CDR-L2 comprising SEQ ID NO:211, and CDR-L3 comprising SEQ ID NO:212. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:63, CDR-H2 comprising SEQ ID NO:64, and CDR-H3 comprising SEQ ID NO:60; the VL region comprises CDR-L1 comprising SEQ ID NO:221, CDR-L2 comprising SEQ ID NO:222, and CDR-L3 comprising SEQ ID NO:223. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:75, CDR-H2 comprising SEQ ID NO:76, and CDR-H3 comprising SEQ ID NO:60; and the VL region comprises CDR-L1 comprising SEQ ID NO:229, CDR-L2 comprising SEQ ID NO:222, and CDR-L3 comprising SEQ ID NO:223. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:52, CDR-H2 comprising SEQ ID NO:53, and CDR-H3 comprising SEQ ID NO:37; the VL region comprises CDR-L1 comprising SEQ ID NO:241, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:242. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:87, CDR-H2 comprising SEQ ID NO:88, and CDR-H3 comprising SEQ ID NO:84; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:247, and CDR-L3 comprising SEQ ID NO:248. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:98, CDR-H2 comprising SEQ ID NO:99, and CDR-H3 comprising SEQ ID NO:84; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:254, and CDR-L3 comprising SEQ ID NO:255. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:108, CDR-H2 comprising SEQ ID NO:109, and CDR-H3 comprising SEQ ID NO:105; the VL region comprises CDR-L1 comprising SEQ ID NO:258, CDR-L2 comprising SEQ ID NO:259, and CDR-L3 comprising SEQ ID NO:260. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:134, CDR-H2 comprising SEQ ID NO:135, and CDR-H3 comprising SEQ ID NO:131; the VL region comprises CDR-L1 comprising SEQ ID NO:246, CDR-L2 comprising SEQ ID NO:273, and CDR-L3 comprising SEQ ID NO:274. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:147, CDR-H2 comprising SEQ ID NO:148, and CDR-H3 comprising SEQ ID NO:144; the VL region comprises CDR-L1 comprising SEQ ID NO:278, CDR-L2 comprising SEQ ID NO:279, and CDR-L3 comprising SEQ ID NO:280. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:160, CDR-H2 comprising SEQ ID NO:161, and CDR-H3 comprising SEQ ID NO:157; the VL region comprises CDR-L1 comprising SEQ ID NO:286, CDR-L2 comprising SEQ ID NO:287, and CDR-L3 comprising SEQ ID NO:288. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:173, CDR-H2 comprising SEQ ID NO:174, and CDR-H3 comprising SEQ ID NO:170; the VL region comprises CDR-L1 comprising SEQ ID NO:294, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:295. In some of any of the 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 of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:173, CDR-H2 comprising SEQ ID NO:174, and CDR-H3 comprising SEQ ID NO:193; the VL region comprises CDR-L1 comprising SEQ ID NO:306, CDR-L2 comprising SEQ ID NO:234, and CDR-L3 comprising SEQ ID NO:307. In some of any of the embodiments, the VH region comprises CDR-H1 comprising SEQ ID NO:160, CDR-H2 comprising SEQ ID NO:161, and CDR-H3 comprising SEQ ID NO:157; and the VL region comprises CDR-L1 comprising SEQ ID NO:311, CDR-L2 comprising SEQ ID NO:312, and CDR-L3 comprising SEQ ID NO:313. In some of any of the 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.

[0008] In some of any of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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 of the 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.

[0009] In some of any of the 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 of the 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 of the 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 of the 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 of the 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 of the 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.

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

[0011] In some of any of the embodiments, the VH region comprises SEQ ID NO:330; and the VL region comprises SEQ ID NO:335. In some of any of the embodiments, the VH region comprises SEQ ID NO:331; and the VL region comprises SEQ ID NO:336. In some of any of the embodiments, the VH region comprises SEQ ID NO:332; and the VL region comprises SEQ ID NO:337. In some of any of the embodiments, the VH region comprises SEQ ID NO:333; and the VL region comprises SEQ ID NO:338. In some of any of the embodiments, the VH region comprises SEQ ID NO:330; and the VL region comprises SEQ ID NO:339. In some of any of the embodiments, the VH region comprises SEQ ID NO:334; and the VL region comprises SEQ ID NO:340.

[0012] In some of any of the embodiments, the PD-L1 protein is a human PD-L1 protein.

[0013] In some of any of the embodiments, the antibody or antigen-binding fragment is recombinant. In some of any of the embodiments, the antibody or antigen-binding fragment is monoclonal. In some of any of the embodiments, the antibody or antigen-binding fragment is a human antibody or antigen-binding fragment, a chimeric antibody or antigen-binding fragment, or a humanized antibody or antigen-binding fragment.

[0014] In some of any of the embodiments, the antibody or antigen-binding fragment comprises a human immunoglobulin Fc region and / or a human antibody framework region.

[0015] In some of any of the embodiments, the antibody or antigen-binding fragment is a single chain antibody fragment. In some of any of the embodiments, the antibody fragment comprises a single chain Fv (scFv).

[0016] In some of any of the embodiments, the antibody is a complete or intact antibody.

[0017] In some of any of the embodiments, the antibody or antigen-binding fragment is a bispecific antibody that further specifically binds to a second antigen. In some of any of the embodiments, the second antigen is an antigen expressed in a tumor cell or an immune cell. In some of any of the embodiments, the second antigen is an antigen expressed in an immune cell that is a T cell, and the antigen is CD25.

[0018] In some of any of the embodiments, the antibody or antigen-binding fragment thereof comprises an Fc region that exhibits one or more Fc-mediated effector functions. In some of any of the embodiments, the antibody or antigen-binding fragment thereof comprises an Fc region that lacks an Fc-mediated effector function, an Fc region that exhibits substantially reduced Fc-mediated effector function, or an Fc region that exhibits no substantial Fc-mediated effector function. In some of any of the embodiments, the antibody or antigen-binding fragment thereof comprises an Fc region that exhibits enhanced Fc-mediated effector function. In some of any of the embodiments, 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).

[0019] In some of any of the embodiments, the antibody or antigen-binding fragment comprises an IgG1 Fc region or an IgG1 isotype, an IgG2 Fc region or an IgG2 isotype, an IgG3 Fc region or an IgG3 isotype, or an IgG4 Fc region or an IgG4 isotype.

[0020] Also provided are conjugates comprising any of the provided antibodies or antigen-binding fragments and a heterologous molecule or moiety. In some of any of the embodiments, the heterologous molecule or moiety is a protein, peptide, nucleic acid, dye, or small molecule. In some of any of the embodiments, the heterologous molecule or moiety is a cytotoxic agent, a toxin, a radioisotope, a chemotherapeutic agent, a lytic peptide, a cytokine, or a photoactivatable dye.

[0021] In some of any of the embodiments, the photoactivatable dye is a phthalocyanine dye. In some of any of the embodiments, the phthalocyanine dye is a Si-phthalocyanine dye.

[0022] In some of any of the embodiments, the phthalocyanine dye is IR700.

[0023] In some of any of the embodiments, the phthalocyanine dye has Formula (I): TIFF2025508739000002.tif61128, or a salt, stereoisomer, or tautomer thereof.

[0024] In some of any of the embodiments, the conjugate is activated to effect cell killing by irradiation at wavelengths at or about 600 nm to 850 nm or about 850 nm.

[0025] In some of any of the embodiments, the activated conjugate provides tumor growth inhibition or killing at a higher level, activity, or potency than the unconjugated antibody.

[0026] In some of any of the embodiments, the antibody or antigen-binding fragment and moiety are linked directly or indirectly via a linker.

[0027] In some of any of the embodiments, the antibody or antigen-binding fragment is covalently attached to a heterologous molecule or moiety.

[0028] In some of any of the embodiments, when contacted with a cell expressing a 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 of the embodiments, when contacted with a cell expressing a PD-L1 protein, the conjugate exhibits reduced internalization compared to an unconjugated antibody or antigen-binding fragment, or a conjugate comprising a reference antibody. In some of any of the embodiments, the reference antibody is avelumab.

[0029] In some of any of the embodiments, the conjugate exhibits substantially reduced binding affinity to 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 to PD-L1 protein of the unconjugated antibody. In some of any of the embodiments, the conjugate exhibits similar binding to PD-L1 protein compared to the unconjugated antibody.

[0030] Polynucleotides encoding any of the provided antibodies or antigen-binding fragments are also provided.

[0031] Also provided are vectors comprising any of the provided polynucleotides. In some of any of the embodiments, the vector is an expression vector.

[0032] Modified cells containing any of the provided vectors are also provided.

[0033] Modified cells expressing any of the provided antibodies or antigen-binding fragments are also provided.

[0034] Compositions comprising any of the provided antibodies or antigen-binding fragments, or any of the provided conjugates, are also provided. In some of any of the embodiments, the compositions also include a pharma- ceutically acceptable excipient.

[0035] Methods of treating a disease or disorder are also provided. Any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions are also provided for use in any of such methods. Use of any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions in the manufacture of a medicament for treating a disease or disorder is also provided. In some of any of the embodiments, the method includes administering any of the provided antibodies or antigen-binding fragments, any of the provided conjugates, or any of the provided compositions to a subject with a disease or disorder.

[0036] Also provided are methods of treatment comprising administering any of the provided compositions to a subject having a disease or disorder. In some of any of the embodiments, the disease or disorder is a tumor or cancer.

[0037] Also provided are methods of treating a tumor or lesion in a subject. In some of any of the embodiments, the method comprises administering to the subject any of the provided conjugates or any of the provided compositions; and irradiating a target area in the subject with 25 J / cm at a wavelength at or about 600 nm to 850 nm at or about 850 nm. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 or at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, thereby activating the conjugate; whereby the growth, volume, or size of the tumor or lesion is reduced or inhibited.

[0038] Methods of treating tumors or lesions are also provided. In some of any of the embodiments, the method comprises administering any of the provided conjugates or any of the provided compositions to a subject having a tumor or lesion that has had a low response to, has been non-responsive, has been resistant, has been refractory, has failed to respond to, or has subsequently relapsed, to a previous immunotherapy; and administering to a target area in which the tumor or lesion is located, at a wavelength of at or about 600 nm to at or about 850 nm, at 25 J / cm. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 or at a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length; wherein the method results in killing of PD-L1 expressing cells in the target area.

[0039] In some of any of the aspects, the previous immunotherapy is treatment with an immune checkpoint inhibitor.

[0040] In some of any of the embodiments, the subject has primary or acquired resistance to previous immunotherapy, including PD-1 / PD-L1 blockade therapy.

[0041] Methods of treating a tumor or lesion are also provided. In some of any of the embodiments, the method comprises administering any of the provided conjugates or any of the provided compositions to a subject who is immune checkpoint inhibitor treatment naive or has not previously been treated with an immune checkpoint inhibitor; and administering to a target area in the subject where the tumor or lesion is located 25 J / cm at a wavelength of at or about 600 nm to at or about 850 nm. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 or at a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length; wherein after irradiation, the growth, size, or viability of the tumor or lesion is reduced or inhibited.

[0042] In some of any of the embodiments, a subject is administered the conjugate to treat, inhibit the growth of, and / or reduce the size of a first tumor or lesion; the method inhibits, delays, or prevents the appearance, growth, or establishment of one or more second tumors or lesions distal to the first tumor or lesion.

[0043] Methods of immunizing a subject having a first tumor or lesion are also provided. In some of any of the embodiments, the method comprises administering any of the provided conjugates to a subject having a tumor or lesion; and irradiating a target area in the first tumor or lesion with 25 J / cm at a wavelength of at or about 600 nm to at or about 850 nm. 2 Or about 25J / cm 2 From 400J / cm2 Or about 400 J / cm 2 or at or about 2 J / cm fiber length to at or about 500 J / cm fiber length; wherein growth of a first tumor or lesion is inhibited and / or reduced in size; and the appearance, growth, or establishment of one or more second tumors or lesions distal to the treated first tumor or lesion is inhibited, delayed, or prevented.

[0044] In some of any of the embodiments, the second tumor or lesion is a metastasis of the first tumor or lesion. In some of any of the embodiments, the method results in the killing of PD-L1 expressing cells in the vicinity of the first tumor or lesion and / or activates an immune cell response, thereby inhibiting, delaying or preventing the appearance, growth or establishment of the second tumor or lesion. In some of any of the embodiments, the second tumor or lesion is phenotypically and / or genotypically the same as the first tumor or lesion. In some of any of the embodiments, the second tumor or lesion is phenotypically and / or genotypically different from the first tumor or lesion.

[0045] In some of any of the embodiments, the second tumor or lesion does not result from a metastasis of the first tumor or lesion. In some of any of the embodiments, the method results in killing of PD-L1 expressing cells or PD-L1 expressing immune cells. In some of any of the 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 of the embodiments, the immune checkpoint protein is selected from among PD-L1, PD-1, and CTLA-4.

[0046] In some of any of the embodiments, the tumor cells do not express PD-L1 in response to an inflammatory stimulus. In some of any of the embodiments, the inflammatory stimulus is interferon.

[0047] In some of any of the embodiments, the tumor cells are not specifically recognized by anti-PD-L1 antibodies. In some of any of the embodiments, the tumor or lesion comprises PD-L1 negative tumor cells. In some of any of the embodiments, at least 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.

[0048] In some of any of the embodiments, the treatment delays regrowth of the tumor or lesion, prevents recurrence of the cancer associated with the tumor or lesion, or extends the period of remission of the cancer associated with the tumor or lesion. In some of any of the embodiments, inhibition of tumor or lesion growth and / or killing of PD-L1 expressing cells is dependent on the presence of CD8+ T cells.

[0049] In some of any of the embodiments, the subject is immune checkpoint inhibitor treatment naive or has not previously been treated with an immune checkpoint inhibitor. In some of any of the embodiments, the subject has been previously treated with an immune checkpoint inhibitor. In some of any of the embodiments, the subject had a low response, was non-responsive, was resistant, was refractory, failed to respond, or subsequently relapsed to a previous treatment with an immune checkpoint inhibitor. In some of any of the embodiments, the inhibition of tumor or lesion growth, size, or viability resulting from the implementation of the method is greater compared to the inhibition resulting from a previous treatment with an immune checkpoint inhibitor.

[0050] In some of any of the embodiments, the immune checkpoint inhibitor is an inhibitor of PD-L1, PD-1, or CTLA-4. In some of any of the embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor. In some of any of the embodiments, the PD-1 inhibitor is an anti-PD-1 antibody. In some of any of the embodiments, the PD-1 inhibitor is an anti-CTLA-4 antibody. In some of any of the embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor. In some of any of the embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody.

[0051] In some of any of the embodiments, the method increases the number or activity of immune cells in the tumor or lesion and / or the microenvironment of the tumor or lesion.

[0052] In some of any of the embodiments, the target area comprises immune cells expressing PD-L1. In some of any of the embodiments, the PD-L1 expressing cells are immune cells. In some of any of the embodiments, the immune cells are monocytes, macrophages, dendritic cells (DCs), or myeloid-derived suppressor cells (MDSCs). In some of any of the embodiments, the immune cells are selected from the group consisting of monocytes, macrophages, e.g., 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 of the embodiments, the immune cells are monocytes. In some of any of the embodiments, the immune cells are macrophages. In some of any of the embodiments, the immune cells are M1 macrophages. In some of any of the embodiments, the immune cells are M2 macrophages. In some of any of the embodiments, the immune cell is an M2 tumor-associated macrophage (M2 TAM). In some of any of the embodiments, the immune cell is a dendritic cell (DC). In some of any of the embodiments, the immune cell is a tolerogenic dendritic cell (tDC). In some of any of the embodiments, the immune cell is a myeloid-derived suppressor cell (MDSC). In some of any of the embodiments, the immune cell is located in the tumor, the tumor microenvironment, or a lymph node.

[0053] In some of any of the embodiments, prior to administration of the conjugate, the subject has a tumor or lesion with a low number or level of CD8+ T cell infiltration. In some of any of the embodiments, after administration and irradiation, the number, level, or activity of immune cells in the tumor or lesion, or in the microenvironment of the tumor or lesion, is increased. In some of any of the embodiments, after administration and irradiation, the number or level of CD8+ T cell infiltration in the tumor or lesion is increased. In some of any of the embodiments, after administration and irradiation, the number or level of memory T cells in the vicinity of the tumor or lesion is increased. In some of any of the embodiments, the targeting molecule is or includes an antibody, antigen-binding antibody fragment, or antibody-like molecule that binds to PD-L1. In some of any of the embodiments, the targeting molecule is or includes an anti-PD-L1 antibody or antigen-binding fragment thereof. In some of any of the embodiments, the target area is a lymph node, or in the vicinity of a lymph node. In some of any of the embodiments, the subject exhibits a durable response, extended progression-free survival, reduced likelihood of recurrence, and / or reduced likelihood of metastasis following administration and irradiation.

[0054] In some of any of the embodiments, the irradiation is performed 30 minutes to 96 hours after administration of the conjugate. In some of any of the embodiments, the irradiation is performed 24 hours ± 4 hours after administration of the conjugate. In some of any of the embodiments, the target area is irradiated with a wavelength of 690 ± 40 nm. In some of any of the embodiments, the target area is irradiated with a wavelength of 670 ± 50 nm. In some of any of the embodiments, the target area is irradiated with 50 J / cm. 2 Or about 50 J / cm 2 or at a dose of 100 J / cm fiber length or about 100 J / cm fiber length.

[0055] In some of any of the embodiments, the tumor, lesion, or cancer is associated with a cancer selected from the group consisting of colon cancer, colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell cancer, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, cancer of the small intestine, spindle cell neoplasm, liver cancer, liver cancer, cancer of the peripheral nerves, brain cancer, cancer of skeletal muscle, cancer of smooth muscle, bone cancer, cancer of adipose tissue, cervical cancer, uterine cancer, cancer of the reproductive organs, lymphoma, and multiple myeloma.

[0056] In some of any of the embodiments, one or more of the steps of the method are repeated. In some of any of the embodiments, the administration of the antibody or antigen-binding fragment, the conjugate, or the composition is repeated one or more times. In some of any of the embodiments, the irradiation step is repeated after each repeated administration of the conjugate or the composition.

[0057] In some of any of the embodiments, the method further comprises administering an additional therapeutic agent or anti-cancer treatment. [Brief description of the drawings]

[0058] [Figure 1-1] Figures 1A-1F show the binding of exemplary anti-PD-L1 antibodies to human PD-L1 (Figures 1A, 1C, and 1E) and cynomolgus monkey PD-L1 (Figures 1B, 1D, and 1F) as determined by ELISA. [Figure 1-2] See description of Figure 1-1. [Figure 1-3] See description of Figure 1-1. [Figure 1-4] Figure 1G shows the binding specificity of exemplary anti-PD-L1 antibody 1P4 to PD-L1 compared to other B7 ligands as determined by ELISA.Figure 1H shows the binding specificity of exemplary anti-PD-L1 antibody 1P9 to PD-L1 compared to other B7 ligands as determined by ELISA. [Diagram 2]Figure 2A shows the binding of exemplary anti-PD-L1 antibodies containing a wild-type or effector knockout Fc region to human PD-L1 expressed on the surface of CHO cells engineered to express human PD-L1, and Figure 2B shows the binding of exemplary anti-PD-L1 antibodies containing a wild-type or effector knockout Fc region to wild-type CHO cells. [Diagram 3] Figure 3 shows the binding of exemplary anti-PD-L1 antibodies to A431 cancer cells that express PD-L1. [Figure 4] Figures 4A-4B show the ADCC activity of exemplary anti-PD-L1 antibodies containing a wild-type Fc region (Figure 4A) or an effector knockout Fc region (Figure 4B). [Diagram 5] Figures 5A-5B show the binding of exemplary anti-PD-L1-IR700 conjugates to human PD-L1 (Figure 5A) and cynomolgus PD-L1 (Figure 5B). [Figure 6] Figure 6A shows the binding of exemplary anti-PD-L1-IR700 conjugates containing a wild-type or effector knockout Fc region to human PD-L1 expressed on the surface of CHO cells engineered to express human PD-L1.Figure 6B shows the binding of exemplary anti-PD-L1-IR700 conjugates containing a wild-type Fc or effector knockout Fc region to wild-type CHO cells. [Figure 7A] Figure 7A shows the binding of an exemplary anti-PD-L1-IR700 conjugate to A431 cancer cells. [Figure 7B] Figure 7B shows binding of an exemplary anti-PD-L1-IR700 conjugate compared to an avelumab-IR700 conjugate to interferon-γ-stimulated A431 cells. [Figure 7C] Figure 7C shows binding of an exemplary anti-PD-L1-IR700 conjugate compared to an avelumab-IR700 conjugate to interferon-γ-stimulated BxPC3 cells. [Figure 7D]Figure 7D shows the binding of an exemplary anti-PD-L1-IR700 conjugate to CHO cells engineered to express human PD-L1 (CHO-hPD-L1 cells) compared to an avelumab-IR700 conjugate. [Figure 7E] Figure 7E shows the binding of exemplary anti-PD-L1-IR700 conjugates and avelumab-IR700 conjugates to wild-type CHO cells. [Figure 8A] Figure 8A shows photoimmunotherapy-induced killing of PD-L1-expressing A431 squamous cell carcinoma cells following irradiation with 1P4-IR700, 1P9-IR700, and avelumab-IR700 conjugates. [Figure 8B] Figures 8B-8C show photoimmunotherapy-induced killing of IFNγ-stimulated A431 cancer cells after 1 hour (Figure 8B) or 24 hours (Figure 8C) of incubation with 1P9-IR700 or avelumab-IR700 conjugates and subsequent irradiation. [Figure 8C] See legend to Figure 8B. [Figure 8D] FIG. 8D shows the viability of human peripheral blood mononuclear cells (PBMCs) from three donors after incubation with 1P9-IR700 or avelumab-IR700 and irradiation. [Figure 8E] FIG. 8E shows the viability of M1 and M2 primary human macrophages after incubation with 1P9-IR700 and irradiation. [Figure 9A] Figures 9A-9C show photoimmunotherapy-induced killing of A431 squamous cell carcinoma cells following irradiation with avelumab-IR700 (Figure 9A), 1P9-IR700 (Figure 9B), and 1P4-IR700 (Figure 9C) at light intensities ranging from 0 J to 128 J. [Figure 9B] See legend to Figure 9A. [Figure 9C] See legend to Figure 9A. [Figure 10A]Figure 10A shows the ability of the exemplary anti-PD-L1 antibody 1P4, the control antibody avelumab, and the exemplary conjugate 1P4-IR700 to block the PD-1 / PD-L1 interaction. [Figure 10B] Figure 10B shows the ability of the exemplary anti-PD-L1 antibody 1P9, the control antibody avelumab, and the exemplary conjugate 1P9-IR700 to block the PD-1 / PD-L1 interaction. [Figure 11] Figure 11 shows the internalization of exemplary anti-PD-L1 conjugates 1P9-IR700 and 1P4-IR700, and the reference conjugate avelumab-IR700, in pancreatic BxPC3 cancer cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] Detailed Description Provided herein are anti-PD-L1 antibodies, anti-PD-L1 antibody fragments, conjugates, compositions, combinations, methods, and uses for treating a subject having a tumor, lesion, or cancer, 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 a subject, such as a 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 generating an enhanced response, e.g., an enhanced response to a second treatment or therapy, in a subject, e.g., a subject having a cancer or tumor, e.g., an invasive cancer, an infiltrating cancer, or a metastatic cancer.

[0060] In some aspects, provided embodiments include administering to a subject an anti-PD-L1 antibody provided herein to directly treat a tumor or lesion, to indirectly treat a tumor or lesion, to enhance systemic immunity, to enhance an immune response, or to enhance the response to a second treatment or therapy.

[0061] In some aspects, the provided embodiments include administering to a subject a conjugate containing any of the provided antibodies or antigen-binding fragments that bind to programmed cell death ligand 1 (PD-L1) conjugated to a phthalocyanine dye, e.g., a silicon phthalocyanine dye, e.g., IR700, or any of the dyes described in WO 2021 / 207691. In some aspects, the provided embodiments include irradiation of a target area, e.g., a target area in which cells expressing PD-L1 are present or may be present. In some aspects, the irradiation results in the death of cells expressing PD-L1 on their surface. In some aspects, the irradiation results in the death of tumor cells expressing PD-L1 on their surface. In some aspects, the irradiation results in the death of myeloid cells expressing PD-L1 on their surface. In some aspects, the irradiation results in the killing of tumor cells and myeloid cells expressing PD-L1 on their surface.

[0062] In some of any of the embodiments provided herein, the provided antibodies, antibody fragments, conjugates, compositions, combinations, methods, and uses are utilized to treat a subject having a tumor, lesion (e.g., a cancerous lesion), or cancer that has had a low response or is substantially non-responsive, has failed, subsequently relapsed, is refractory, and / or is resistant to a previous therapeutic treatment, e.g., a previous immunomodulatory agent treatment and / or a previous anti-cancer therapeutic treatment.

[0063] In some aspects, anti-PD-L1 antibodies, and in some cases, additional therapeutic agents, are utilized in the provided compositions, combinations, methods, and uses. In some aspects, phthalocyanine dye-targeting molecule conjugates (e.g., anti-PD-L1 conjugated to IR700), and in some cases, additional therapeutic agents, are utilized in the provided compositions, combinations, methods, and uses. Uses include the use of antibodies, antibody fragments, conjugates, compositions, and combinations in such methods, e.g., therapeutic methods, and treatments, e.g., treatment regimens, and the use of such antibodies, antibody fragments, conjugates, compositions, and combinations in the preparation of medicaments for carrying out such therapeutic methods and treatments. Also provided are such antibodies, antibody fragments, conjugates, compositions, and combinations for use in the treatment of tumors, lesions, or cancers. In some aspects, such uses include carrying out a method or treatment described herein, e.g., any therapeutic method or treatment regimen. In some aspects, the method and use also include irradiating a target area, for example, a target area in which a tumor, a lesion, or a cancer is located in a subject, with light, for example, as described herein. In some embodiments, the method and use thereby treats the tumor, the lesion, or the cancer. In some aspects, the tumor, the lesion, or the cancer to be treated includes, for example, a cancer in a subject that includes a primary tumor and a secondary or metastatic tumor cell, for example, a secondary or metastatic cancer. In some aspects, the tumor, the lesion, or the cancer can include a primary tumor or multiple primary tumors, and a metastatic tumor cell. In some cases, the subject to be treated can have one or more of a primary tumor, a metastatic tumor cell, and / or an invasive tumor cell.

[0064] In some aspects, methods and uses of such antibodies, antibody fragments, conjugates, compositions, and combinations in enhancing, activating, inducing, eliciting, strengthening, or supporting immune function, such as local and / or systemic immunity, in a subject are also provided. In some aspects, the provided embodiments may target tumor cells. In some aspects, the provided embodiments may target cells of the tumor microenvironment, such as non-cancerous cells and / or immune cells, such as antigen-presenting cells, or myeloid cells with immune suppressive function.

[0065] One of the challenges in treating cancer patients is the lack of responsiveness to cancer therapeutics. There is an urgent need for compositions and methods for treating such cancers. In some circumstances, the provided embodiments are based on the observation that treatment with the anti-PD-L1 antibodies or antibody fragments described herein results in a significant reduction in PD-1:PD-L1 interaction. Such blocking of PD-1:PD-L1 interaction can lead to substantial inhibition of tumor growth and / or a complete response to treatment with the antibody or antibody fragment. In some aspects, the exemplary anti-PD-L1 antibodies or antibody fragments provided herein were observed to 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 the antibody and reduced inhibition by blocking the PD-1:PD-L1 interaction.

[0066] In some aspects, the provided anti-PD-L1 antibodies or antibody fragments also provide the advantage of retaining binding affinity after conjugation with an additional agent, such as a phthalocyanine dye. In contrast, conjugates comprising the same agent (e.g., a phthalocyanine dye) and a reference anti-PD-L1 antibody were observed to show a substantial reduction in binding affinity, e.g., an almost 50-fold reduction. Such an effect of retaining binding affinity after conjugation of the provided antibodies is unexpected and advantageous. In some aspects, the provided antibodies retain their activity, e.g., binding affinity, even when additional agents are conjugated, which supports the availability and advantages of antibodies for use in generating conjugates, e.g., antibody-drug conjugates and phthalocyanine dye-antibody conjugates, and the use of the conjugates, e.g., therapeutic use, experimental use, or imaging. The results described herein also demonstrate the advantages of the provided antibodies and antibody fragments, and conjugates comprising the provided antibodies and antibody fragments, such as antibody-phthalocyanine dye conjugates, in therapeutic applications, such as photoimmunotherapy (PIT).

[0067] In some circumstances, the provided embodiments are based on the observation that treatment 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 light irradiation of the target area (also referred to as "photoimmunotherapy" and "PIT"), results in substantial inhibition of tumor growth and / or a complete response to treatment.

[0068] In some aspects, treatment with the anti-PD-L1 antibodies provided herein can activate, induce, enhance, or potentiate the immune response, for example, by binding to PD-L1 expressed on immunosuppressive cells, e.g., immunosuppressive myeloid cells (e.g., myeloid-derived suppressor cells (MDSC), tolerogenic dendritic cells (tDC), M1 macrophages, M2 tumor-associated macrophages (M2 TAM)), thereby eliminating or reducing PD-L1:PD-1 interactions. 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 the immune response, for example, by eliminating immunosuppressive cells, e.g., immunosuppressive myeloid cells (e.g., myeloid-derived suppressor cells (MDSC), tolerogenic dendritic cells (tDC), M1 macrophages, M2 tumor-associated macrophages (M2 TAM)). In some aspects, the elimination of immunosuppressive cells results in the activation, induction, enhancement, or strengthening of an immune response, for example, an anti-tumor or anti-cancer immune response. In some aspects, any of the provided embodiments provide the advantage that they can be applied to many different tumors, lesions, or cancer types, for example, 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 utilized to overcome such immunosuppressive mechanisms.

[0069] Furthermore, in some aspects, the provided embodiments may provide effective treatment of tumors, lesions, or cancers that are heterogeneous, e.g., contain a variety of different types of tumor cells or cancer cells. In some aspects, the provided embodiments also provide the advantage of inducing, activating, or enhancing the local and / or systemic immune activity or systemic immunity of a subject, allowing treatment of tumors, lesions, or cancers that are present elsewhere in the body other than the irradiation target area, e.g., metastatic tumors or cancers, invasive tumors or cancers, tumors or cancers at different sites, or tumors, lesions, or cancers of different types. Other advantages include the treatment of metastatic and / or invasive cancers without the need to locate and / or directly irradiate metastatic tumor cells.

[0070] The provided embodiments may also be used to treat tumors, lesions, or cancers that are not responsive to previous therapeutic treatments, e.g., immune checkpoint inhibitors, anti-cancer drugs, or molecules against immune suppressive cells. The provided embodiments also provide other advantages in the treatment of cancer, e.g., effective treatment of cancers that are not responsive to previous therapeutic treatments, e.g., other anti-PD-L1 treatments.

[0071] The present disclosure also provides unexpected features in enhancing anti-cancer or anti-tumor immunity in a subject, for example, against different tumors or cancers that may develop. The provided embodiments are based on the observation that in some circumstances, treatment of cancer with a phthalocyanine dye-anti-PD-L1 antibody or antibody fragment conjugate, for example, an anti-PD-L1 antibody-IR700 conjugate, and subsequent irradiation of the tumor not only results in treatment of that particular tumor, but also results in effective treatment of tumors that develop subsequently, of the same or different type. The provided embodiments also provide effective treatment of tumors that are introduced after the subject has had a complete response after treatment of the first tumor, which shows an immune memory response; and / or effective treatment of tumors that are distal to the target area of ​​irradiation (e.g., metastasized tumors or tumors that exist in different locations). The provided compositions, combinations, methods, and uses may result in an enhancement or improvement of a subject's immune response, for example, a systemic immune response against cancer, for example, an immune memory response that may be effective against tumors that may develop after treatment.

[0072] In some aspects, methods are also provided that include administration of an additional therapeutic agent, such as an immunomodulatory agent, in combination with the anti-PD-L1 antibody or fragment, or phthalocyanine dye conjugate thereof (e.g., an anti-PD-L1-IR700 conjugate).

[0073] In some of any of the embodiments provided, treatment with or administration of the anti-PD-L1 conjugate is generally followed by irradiation with light of an appropriate wavelength. Such irradiation is considered part of the treatment and administration of the anti-PD-L1 conjugates provided herein, unless it is specifically stated that an irradiation step is not performed in the method. In some cases, such irradiation is referred to as photoimmunotherapy (PIT). In some circumstances, the exemplary anti-PD-L1 antibodies, antibody fragments, or conjugates provided herein have been observed to exhibit increased or decreased internalization compared to unconjugated antibodies or antigen-binding fragments, or conjugates comprising a reference antibody. In some circumstances, the exemplary anti-PD-L1 antibodies, antibody fragments, or conjugates provided herein have been observed to exhibit increased internalization compared to unconjugated antibodies or antigen-binding fragments, or conjugates comprising a reference antibody.

[0074] All publications, e.g., patent documents, scientific literature, and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated by reference. To the extent that the definitions set forth herein contradict or are otherwise inconsistent with the definitions set forth in the patents, applications, published applications, and other publications incorporated herein by reference, the definitions set forth herein take precedence over the definitions incorporated herein by reference.

[0075] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0076] I. PD-L1-Binding Molecules and Polynucleotides Encoding The Same In some aspects, PD-L1 binding molecules, e.g., PD-L1 binding polypeptides, are provided. Such binding molecules include antibodies (e.g., antigen-binding fragments) that specifically bind to PD-L1 protein, e.g., human PD-L1 protein (huPD-L1). Also included among the binding molecules are polypeptides that contain such antibodies, e.g., multispecific antibodies, e.g., bispecific antibodies, that bind to PD-L1 in addition to one or more other antigens.

[0077] 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). PD-L1 expression can be induced in dendritic cells and keratinocytes by IFNγ stimulation. PD-L1 is expressed in activated T cells, B cells, myeloid 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 (M2 TAMs), tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs), or on certain tumor cells, to induce immune suppression in the vicinity of the tumor or tumor microenvironment (TME).

[0078] PD-L1 is the cognate ligand of PD-1. The complex of PD-1 and PD-L1 inhibits the proliferation of CD8+ T cells and reduces immune responses (Topalian et al., 2012, N Engl J Med 366:2443-54; Brahmer et al., 2012, N Engl J Med 366:2455-65). The primary 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 binding of PD-1 to one of its endogenous ligands, e.g., PD-L1, acts to inhibit T cell activation by inhibiting stimulatory kinases (Pardoll, 2012, Nature Reviews Cancer 12:252-264). PD-1 also acts to inhibit TCR "stop signals" (Pardoll, 2012, Nature Reviews Cancer 12:252-264). PD-1 is highly expressed on regulatory T (Treg) cells and can increase proliferation in the presence of ligand (Pardoll, 2012, Nature Reviews Cancer 12:252-264). Binding of PD-L1 to PD-1 transmits an inhibitory signal based on interaction with phosphatases (SHP-1 or SHP-2) via the immunoreceptor tyrosine-based switch motif (ITSM).

[0079] Thus, the PD-L1:PD-1 pathway functions as a checkpoint in the negative control of some immune responses and is a major immunoinhibitory mediator of T cell exhaustion. Blockade of this pathway can lead to enhanced T cell activation, expansion, and effector function (Sakuishi et al., JEM Vol. 207, September 27, 2010, pp2187-2194).

[0080] PD-L1 is also expressed in tumor cells. Tumors known to overexpress PD-L1 include, but are not limited to, those causing breast cancer, lung cancer, bladder cancer, colon 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 hematological malignancies (Brahmer et al., N Engl 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 & 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.

[0081] Human PD-L1 is expressed as a 290 amino acid (aa) type I membrane precursor protein with a putative 18 aa signal peptide, a 221 aa extracellular domain, a 21 aa transmembrane region, and a 31 aa cytoplasmic domain. For example, provided are antibodies and antigen-binding fragments thereof that specifically bind to PD-L1, e.g., human PD-L1, as well as conjugates and molecules comprising such antibodies or antigen-binding fragments.

[0082] A. Anti-PD-L1 antibody Anti-PD-L1 antibodies, e.g., functional antigen-binding fragments, are provided. In some embodiments, the antibodies or antigen-binding fragments contain a heavy chain variable region (VH) and a light chain variable region (VL) that together are capable of specifically binding to PD-L1. In some embodiments, the antibodies or antigen-binding fragments include full-length IgG molecules, or fragments, e.g., scFv, Fab, F(ab), that specifically bind to PD-L1, e.g., human PD-L1. 2 Among the anti-PD-L1 antibodies provided are human antibodies. Antibodies include isolated antibodies.

[0083] The term "antibody" is used herein in its broadest sense and includes polyclonal and monoclonal antibodies, e.g., intact antibodies, as well as functional (antigen-binding) antibody fragments, e.g., fragment antigen binding (Fab) fragments, F(ab') fragments, and the like. 2 The term includes fragments, Fab fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (VH) regions capable of specifically binding to an antigen, single chain antibody fragments, e.g., single chain variable fragments (scFv), and single domain antibody (e.g., sdAb, sdFv, nanobody) fragments. The term includes genetically engineered and / or otherwise modified forms of immunoglobulins, e.g., 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 indicated, the term "antibody" should be understood to encompass functional antibody fragments thereof, also referred to herein as "antigen-binding fragments". The term also includes intact or full-length antibodies, e.g., antibodies of any class or subclass, e.g., IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0084] The terms "complementarity determining region" and "CDR", synonymous with "hypervariable region" or "HVR", are known in the art to refer to discontinuous amino acid sequences in antibody variable regions 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. The terms "framework region" and "FR" are known in the art to refer to the non-CDR portions of the heavy and light chain variable regions. 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.

[0085] The exact amino acid sequence boundaries of a given CDR or FR may be determined according to any of a number of well-known schemes, e.g., Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of 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 site topography," J. Mol. Biol. 262, 732-745." ("Contact" numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003, 27(1):55-77 (the "IMGT" numbering scheme); Honegger A and Pluckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001, 309(3):657-70 (the "Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272 (the "AbM" numbering scheme).

[0086] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. For both the Kabat scheme and the Chothia scheme, the numbering is based on the most common antibody region sequence length, and in some antibodies, insertions and deletions are accommodated by the insertion letter, e.g., "30a". These two schemes place certain insertions and deletions ("indels") in different positions, resulting in differential numbering. The Contact scheme is based on the analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on the one used by Oxford Molecular's AbM antibody modeling software.

[0087] Table 1 below lists exemplary position 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. Residue numbering for CDR-H1 is described using both the Kabat and Chothia numbering schemes. FRs are located between the CDRs, e.g., FR-L1 in front of CDR-L1, FR-L2 between CDR-L1 and CDR-L2, FR-L3 between CDR-L2 and CDR-L3, and so on. Note that the Kabat numbering scheme shown places the insertion at H35A and H35B, so the end of the Chothia CDR-H1 loop when numbered using the Kabat numbering rules shown varies from H32 to H34, depending on the length of the loop.

[0088] Table 1. CDR boundaries according to various numbering schemes TIFF2025508739000003.tif511611 - Kabat et al.(1991),"Sequences of Proteins of Immunological Interest,"5th Ed.Public Health Service,National Institutes of Health,Bethesda,MD 2 - Al-Lazikani et al.,(1997)JMB 273,927-948

[0089] Thus, unless otherwise specified, the "CDRs" or "complementarity determining regions" of a given antibody or region thereof, such as a variable region, or individual designated CDRs (e.g., CDR-H1, CDR-H2, CDR-H3), should be understood to encompass the complementarity determining regions (or specific complementarity determining regions) defined by any of the above or other known schemes. For example, when a particular CDR (e.g., CDR-H3) is noted as containing the amino acid sequence of a corresponding CDR in the amino acid sequence of a given VH or VL region, it is understood that such CDR has the sequence of the corresponding CDR (e.g., CDR-H3) in the variable region defined by any of the above or other known schemes. In some embodiments, when an antibody or antigen-binding fragment thereof is described as comprising CDR-H1, CDR-H2, and CDR-H3 contained within a given VH region amino acid sequence, and CDR-L1, CDR-L2, and CDR-L3 contained within a given VL region amino acid sequence, the CDRs may be defined according to any of the above schemes, such as the Kabat, Chothia, AbM, IMGT, or Contact methods, or other known schemes. In some embodiments, a specific CDR sequence is specified. Although the exemplary CDR sequences of the provided antibodies are described using various numbering schemes (see, for example, Tables E1a and E1b), it is understood that the provided antibodies may comprise CDRs described according to any of the other above numbering schemes or other numbering schemes known to those skilled in the art.

[0090] Similarly, unless otherwise specified, a given antibody or region thereof, e.g., a FR of a variable region or an individual designated FR (e.g., FR-H1, FR-H2, FR-H3, FR-H4, FR-L1, FR-L2, FR-L3, and / or FR-L4), should be understood to encompass framework regions (or specific framework regions) defined by any of the known schemes. In some situations, a scheme for the identification of a specific CDR or FR is specified, e.g., CDRs are defined by the Kabat, Chothia, AbM, IMGT, or Contact methods, or other known schemes. In other cases, the specific amino acid sequences of the CDRs or FRs are given. In some embodiments, when an antibody or antigen-binding fragment thereof is described as comprising FR-H1, FR-H2, FR-H3, and FR-H4 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 FRs may be defined by any of the schemes described above, e.g., the Kabat, Chothia, AbM, IMGT, or Contact methods, or other known schemes.

[0091] The term "variable region" or "variable domain" refers to the domain of an antibody's heavy or light chain that is involved in binding the antibody to an antigen. The variable regions of the heavy and light chains of native antibodies (VH and VL, respectively) generally have similar structures, with each domain containing four conserved framework regions (FR) and three CDRs (see, for example, Kindt et al. Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007)). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a particular antigen may be isolated by using a VH or VL domain derived from an antibody that binds the antigen, respectively, to screen a library of complementary VL or VH domains. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0092] Among the antibodies provided are antibody fragments. 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 Fv, Fab, Fab', Fab'-SH, F(ab') 2 ; diabody; linear antibody; variable heavy chain (V H ) regions, single chain antibody molecules, e.g., scFv, and V H In a specific embodiment, the antibody comprises a variable heavy chain (V H ) and variable light chain (V L In some embodiments, the antibody is a single chain antibody fragment, e.g., an scFv, comprising a heavy chain variable (VH) region and / or a light chain variable (VL) region.

[0093] 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, the antibody is a recombinantly produced fragment, such as a fragment that includes a non-naturally occurring arrangement, such as two or more antibody regions or chains joined by a synthetic linker, such as a peptide linker, and / or cannot be produced by enzymatic digestion of naturally occurring intact antibodies. In some aspects, the antibody fragment is an scFv.

[0094] A "humanized" antibody is an antibody in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A humanized antibody may optionally contain 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 variant of a non-human antibody that has undergone humanization, typically to reduce immunogenicity to humans, while retaining the specificity and affinity of the non-human parent antibody. In some embodiments, some FR residues of a humanized antibody are replaced with the corresponding residues derived from a non-human antibody (e.g., the antibody from which the CDR residues are derived), e.g., to restore or improve the specificity or affinity of the antibody.

[0095] Among the anti-PD-L1 antibodies provided are human antibodies. A "human antibody" is an antibody produced by a human or human cell, or a non-human source utilizing a human antibody repertoire, or other human antibody coding sequence, e.g., an antibody having an amino acid sequence corresponding to that of a human antibody library. Excluded from the term are humanized versions of non-human antibodies that contain non-human antigen-binding regions, e.g., those in which all or substantially all CDRs are non-human. The term includes antigen-binding fragments of human antibodies.

[0096] Human antibodies can be prepared by administering immunogens to transgenic animals that have been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. Such animals typically contain all or part of a human immunoglobulin locus that replaces the endogenous immunoglobulin locus, or that is extrachromosomally present or randomly integrated into the animal's chromosomes. In such transgenic animals, the endogenous immunoglobulin locus is generally inactivated. Human antibodies can also be derived from human antibody libraries, such as phage display libraries and cell-free libraries that contain antibody coding sequences derived from human repertoires.

[0097] Among the antibodies provided are monoclonal antibodies, e.g., monoclonal antibody fragments. The term "monoclonal antibody" as used herein refers to an antibody obtained from or within a population of substantially homogeneous antibodies, i.e., the individual antibodies constituting the population are identical, except for variants that may contain naturally occurring mutations or that may arise during the production of the monoclonal antibody preparation, and such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically contain different antibodies against different epitopes, each monoclonal antibody of a monoclonal antibody preparation is directed against a single epitope on an antigen. This term should not be construed as requiring the production of the antibody by a particular method. Monoclonal antibodies can be produced by a variety of techniques, including, but not limited to, production from hybridomas, recombinant DNA methods, phage display, and other antibody display methods.

[0098] The terms "polypeptide" and "protein" are used interchangeably to refer to a polymer of amino acid residues and are not limited to a minimum length. Polypeptides, including the provided antibodies and antibody conjugates, may contain amino acid residues, e.g., natural amino acid residues and / or non-natural amino acid residues. These terms also include post-translational modifications of the polypeptide, e.g., glycosylation, sialylation, acetylation, phosphorylation, and the like. In some aspects, the polypeptide may contain modifications to the native or natural sequence, so long as the protein maintains a desired activity. These modifications may be deliberate, e.g., by site-directed mutagenesis, or may be accidental, e.g., by mutation of the host that produces the protein or by errors due to PCR amplification.

[0099] 1. Exemplary Antibodies 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), or a sufficient antigen-binding portion thereof, as described herein. 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, that contains 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, that contains 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 that contains CDR-H1, CDR-H2, and / or CDR-H3 as described herein, and contains a VL region sequence that contains CDR-L1, CDR-L2, and / or CDR-L3 as described herein. Among the antibodies provided are those that have sequences at least 90%, or at least 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.

[0100] In some embodiments, the antibody or antigen-binding fragment thereof has a heavy chain variable (VH) region having a sequence selected from any 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 sequence selected from any 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.

[0101] In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any of SEQ ID NOs:1-11, or a sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region amino acid sequence selected from any of SEQ ID NOs:1-11, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence.

[0102] In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any of SEQ ID NOs:12-16, or a sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region amino acid sequence selected from any of SEQ ID NOs:12-16, or contains CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence.

[0103] In some embodiments, the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment having a VH region that includes CDR-H1, CDR-H2, and / or CDR-H3 derived from a VH region selected from any of SEQ ID NOs: 12-16. In some embodiments, the antibody or antigen-binding fragment thereof has a VH having a sequence selected from any of SEQ ID NOs: 330-334, or a sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region amino acid selected from any of SEQ ID NOs: 330-334, or contains the CDR-H1, CDR-H2, and / or CDR-H3 present in such a VH sequence.

[0104] In some embodiments, the antibody or antigen-binding fragment thereof has a VH region having a sequence selected from any 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 to the VH region amino acids selected from any 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 to SEQ ID NO:1, 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 to SEQ ID NO:2, 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 to SEQ ID NO:6, 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.

[0105] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any of SEQ ID NOs:1-16. In some of such embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Chothia numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the AbM numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Contact numbering. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the IMGT numbering.

[0106] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any of SEQ ID NOs:1-11. In some of such embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Chothia numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the AbM numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Contact numbering. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the IMGT numbering.

[0107] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any of SEQ ID NOs:12-16. In some of such embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Chothia numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the AbM numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Contact numbering. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the IMGT numbering.

[0108] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 present in a VH comprising a sequence selected from any of SEQ ID NOs:330-334. In some of such embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Chothia numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the AbM numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and / or CDR-H3 according to the Contact numbering. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the IMGT numbering.

[0109] In some embodiments, the VH region of the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the Chothia numbering shown in Table E1a. In some embodiments, the VH region of the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the AbM numbering shown in Table E1a. In some embodiments, the VH region of the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering shown in Table E1a. In some embodiments, the VH region of the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the Contact numbering shown in Table E1a. In some embodiments, the VH region of the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the IMGT numbering shown in Table E1a.

[0110] In some embodiments, the provided antibody or antigen-binding fragment thereof comprises 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, according to the Chothia numbering. CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 48, 95, and 84, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 48, 104, and 105, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 116, 117, and 118, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 129, 130, and 131, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 142, 143, and 144, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs: 155, 156, and 157, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs:168, 169, and 170, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs:35, 181, and 182, respectively; CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs:168, 169, and 193, respectively; and CDR-H1, CDR-H2, and CDR-H3 comprising the sequences of SEQ ID NOs:197, 198, and 199, respectively.

[0111] For example, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence selected from among 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 the Chothia numbering system.

[0112] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3, each comprising a sequence selected from among 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, according to the 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, each comprising SEQ ID NOs: 35, 36, and 37, according to the 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, each comprising SEQ ID NOs: 82, 83, and 84, according to the Chothia numbering.

[0113] 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, according to the 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, according to the Kabat numbering.

[0114] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any of SEQ ID NOs: 1-16. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any of SEQ ID NOs: 1-11. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any of SEQ ID NOs: 12-16. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence selected from any of SEQ ID NOs:330-334. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence of SEQ ID NO:1. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-H1, CDR-H2, and CDR-H3, each of which comprises a sequence of CDR-H1, CDR-H2, and CDR-H3 contained within a VH region sequence of SEQ ID NO:2. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence of SEQ ID NO:6.In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, and CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence of SEQ ID NO:7.

[0115] Also provided are antibodies and antigen-binding fragments thereof having sequences at least, or at least about, 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 having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VH region sequence selected from any of SEQ ID NOs: 1-16 and 330-334.

[0116] In some embodiments, the antibody is a single domain antibody (sdAb) that includes only a VH region sequence or a sufficient antigen-binding portion thereof, such as any of the VH sequences set forth above (e.g., CDR-H1, CDR-H2, and / or CDR-H3).

[0117] In some embodiments, an antibody (e.g., an anti-PD-L1 antibody) or antigen-binding fragment thereof provided herein comprising a VH region further comprises a light chain or a 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 a sufficient antigen-binding portion of a VH region and a 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, e.g., a Fab or scFv. In some such embodiments, the antibody is a full-length antibody that also contains a constant region.

[0118] In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any 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 of SEQ ID NOs: 17-34 and 335-340, or contains CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0119] In some embodiments, the antibody or antigen-binding fragment thereof has a VL having a sequence selected from any 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 selected from any of SEQ ID NOs:17-28, or contains CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0120] In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any 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 selected from any of SEQ ID NOs:29-34, or contains CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0121] In some embodiments, the antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment having a VL region that includes CDR-L1, CDR-L2, and / or CDR-L3 derived from a VL region selected from any of SEQ ID NOs:29-34. In some embodiments, the antibody or antigen-binding fragment thereof has a light chain variable (VL) region having a sequence selected from any of SEQ ID NOs: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 selected from any of SEQ ID NOs:335-340, or contains the CDR-L1, CDR-L2, and / or CDR-L3 present in such a VL sequence.

[0122] In some embodiments, the antibody or antigen-binding fragment thereof has a VL having a sequence selected from any 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 selected from any of SEQ ID NOs:21-24, 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:21, or a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO:21, 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: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 the 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 the 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.

[0123] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Chothia numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the AbM numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Contact numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the IMGT numbering of a VL comprising a sequence selected from any of SEQ ID NOs:17-34.

[0124] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Chothia numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the AbM numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Contact numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the IMGT numbering of a VL comprising a sequence selected from any of SEQ ID NOs:17-28.

[0125] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Chothia numbering of a VL comprising a sequence selected from any of SEQ ID NOs:29-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the AbM numbering of a VL comprising a sequence selected from any of SEQ ID NOs:29-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering of a VL comprising a sequence selected from any of SEQ ID NOs:29-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Contact numbering of a VL comprising a sequence selected from any of SEQ ID NOs:29-34. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the IMGT numbering of a VL comprising a sequence selected from any of SEQ ID NOs:29-34.

[0126] In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Chothia numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the AbM numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the provided antibodies or antigen-binding fragments thereof comprise CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the Contact numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the IMGT numbering of a VL comprising a sequence selected from any of SEQ ID NOs: 335-340.

[0127] In some embodiments, the VL region of the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the Chothia numbering as shown in Table E1b. In some embodiments, the VL region of the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the AbM numbering as shown in Table E1b. In some embodiments, the VL region of the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering as shown in Table E1b. In some embodiments, the VL region of the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the Contact numbering as shown in Table E1b. In some embodiments, the VL region of the provided antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the IMGT numbering as shown in Table E1b.

[0128] In some embodiments of the antibodies or antigen-binding fragments thereof provided herein, the VL region comprises 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: 241, 234, and 242, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 246, 247, and 248, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 246, 254, and 255, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 258, 259, and 260, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 265, 266, and 267, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 246, 273, and 274, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 278, 279, and 280, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 286, 287, and 288, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 294, 234, and 295, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 299, 300, and 301, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 306, 234, and 307, respectively;CDR-L1, CDR-L2, and CDR-L3 selected from CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 311, 312, and 313, respectively; CDR-L1, CDR-L2, and CDR-L3 comprising the sequences of SEQ ID NOs: 319, 320, and 321, respectively;

[0129] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein are 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; and SEQ ID NOs:258, 259, and 260. 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 sequences selected from among SEQ ID NOs:278, 279, and 280, respectively. 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 sequences selected from among SEQ ID NOs:233, 234, and 235, respectively, according to the 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 sequences selected from among SEQ ID NOs:246, 247, and 248, respectively, according to the Chothia numbering.

[0130] 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 the 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 the Kabat numbering.

[0131] 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 a sequence selected from among SEQ ID NOs:233, 234, and 235; SEQ ID NOs:241, 234, and 242; SEQ ID NOs:246, 247, and 248; and SEQ ID NOs:246, 254, and 255 according to Kabat numbering, Chothia numbering, or AbM numbering.

[0132] In some embodiments, the provided antibodies or antigen-binding fragments thereof contain CDR-L1, CDR-L2, and CDR-L3, each contained within a VL region sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof contain CDR-L1, CDR-L2, and CDR-L3, each contained within a VL region sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibodies or antigen-binding fragments thereof contain CDR-L1, CDR-L2, and CDR-L3, each contained within a VL region sequence selected from any of SEQ ID NOs: 29-34. In some embodiments, the provided antibodies or antigen-binding fragments thereof contain CDR-L1, CDR-L2, and CDR-L3, each contained within a VL region sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the provided antibodies contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO:21. In some embodiments, the provided antibodies contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO:22. In some embodiments, the provided antibodies contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO:23. In some embodiments, the provided antibodies contain CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence of SEQ ID NO:24.

[0133] In some embodiments, the provided antibody or antigen-binding fragment thereof comprises a VL region comprising a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises a VL region comprising a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the provided antibody or antigen-binding fragment thereof comprises a VL region comprising a sequence selected from any of SEQ ID NOs: 29-34. In some embodiments, the provided antibody or antigen-binding fragment thereof contains a VL region comprising a sequence of SEQ ID NO: 21. In some embodiments, the provided antibody or antigen-binding fragment thereof contains a VL region comprising a sequence of SEQ ID NO: 22. In some embodiments, the provided antibody or antigen-binding fragment thereof contains a VL region comprising a sequence of SEQ ID NO: 23. In some embodiments, the provided antibody or antigen-binding fragment thereof contains a VL region comprising a sequence of SEQ ID NO: 24. Antibodies having a sequence at least, or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of such sequences are also provided.

[0134] In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 1-16, and the VL region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 17-34. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 1-11, and the VL region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 17-28. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 330-334, and the VL region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 335-340. In some embodiments, the VH region of the antibody or fragment comprises a sequence selected from any of SEQ ID NOs: 1, 2, 6, and 7, and the VL region of the antibody or fragment comprises a sequence selected from any 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.

[0135] Antibodies and antigen-binding fragments thereof having sequences at least, or at least about, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to such sequences are also provided. For example, provided herein are antibodies or antigen-binding fragments containing 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 of SEQ ID NOs:1-16 and / or 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 of SEQ ID NOs:17-34. 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 of SEQ ID NOs: 1-11, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any of SEQ ID NOs: 17-28. 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 of SEQ ID NOs:330-334, and a VL region having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a VL region sequence selected from any 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 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 VL region sequence selected from any of SEQ ID NOs:21-24. 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 the sequence set forth 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 set forth in SEQ ID NO:21. 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 the sequence set forth 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 set forth in SEQ ID NO:22. 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 the sequence set forth 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 set forth in SEQ ID NO:23.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 the sequence set forth 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 set forth in SEQ ID NO:24.

[0136] 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 of SEQ ID NOs:1-16; and a VL having at least 90% sequence identity to a VL sequence selected from any of SEQ ID NOs:17-34.

[0137] 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 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 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 the VH sequence of SEQ ID NO:1; and a VL having at least 90% sequence identity to the 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 the VH sequence of SEQ ID NO:2; and a VL having at least 90% sequence identity to the 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 the VH sequence of SEQ ID NO:6; and a VL having at least 90% sequence identity to the 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.

[0138] In some embodiments, the VH region of the antibody or antigen-binding fragment thereof comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within a VH region sequence selected from any of SEQ ID NOs:1-16; and the VL region of the antibody or antigen-binding fragment thereof comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within a VL region sequence selected from any of SEQ ID NOs:17-34.

[0139] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within a VH region sequence selected from any of SEQ ID NOs:1-11, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within a VL region sequence selected from any of SEQ ID NOs:17-28.

[0140] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within a VH region sequence selected from any of SEQ ID NOs:12-16, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within a VL region sequence selected from any of SEQ ID NOs:29-34.

[0141] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within a VH region sequence selected from any of SEQ ID NOs:330-334, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within a VL region sequence selected from any of SEQ ID NOs:335-340.

[0142] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within a VH region sequence selected from any of SEQ ID NOs:1, 2, 6, and 7, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within a VL region sequence selected from any of SEQ ID NOs:21-24.

[0143] In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:1, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:17. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:2, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:18. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:3, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:19. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:4, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:19.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:5, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:20. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:1, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:21. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:2, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:22. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:6, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:23.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:7, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:24. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:8, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:25. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:9, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:26. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:10, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:27.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:11, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:28. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:12, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:29. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:13, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:30. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:14, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:31.In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:15, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:32. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:12, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:33. In some embodiments, the VH region of the antibody or fragment comprises CDR-H1, CDR-H2, CDR-H3, which comprise the sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the VH region sequence set forth in SEQ ID NO:16, and the VL region of the antibody or fragment comprises CDR-L1, CDR-L2, CDR-L3, which comprise the sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, contained within the VL region sequence set forth in SEQ ID NO:34.

[0144] In some embodiments, the antibody or antigen-binding fragment thereof is a VH comprising CDR-H1, CDR-H2, and CDR-H3, wherein CDR-H1 is selected from the group consisting of SEQ ID NOs: 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, 132, 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, and 204, and 207; NO:36, 39, 41, 43, 46, 49, 51, 53, 55, 57, 59, 62, 64, 66, 69, 72, 74, 76, 78, 80, 83, 86, 88, 9 0, 93, 95, 97, 99, 101, 103, 104, 107, 109, 111, 114, 117, 120, 122, 124, 127, 130, 133, 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; A VH comprising a sequence selected from any 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 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; 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; 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;

[0145] In some embodiments, CDR-H1 comprises a sequence selected from any 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 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; CDR-H3 comprises a sequence selected from any of SEQ ID NOs: 37, 44, 47, 84, 91, and 94; and CDR-L1 comprises a sequence selected from any of SEQ IDs: CDR-L1 comprises a sequence selected from any of SEQ ID NOs: 233, 236, 239, 241, 243, 245, 246, 249, and 252; CDR-L2 comprises a sequence selected from any of SEQ ID NOs: 217, 234, 237, 240, 247, 250, 253, 254, and 256; and CDR-L3 comprises a sequence selected from any of SEQ ID NOs: 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; CDR-H3 comprises the sequence of SEQ ID NO:37; 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; CDR-H3 comprises the sequence of SEQ ID NO:84; 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.

[0146] 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:216, 217, and 219, respectively. and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 221, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 58, 59, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 221, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 71, 72, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 229, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 71, 72, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 229, 222, and 223, respectively. and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs:35, 36, and 37, respectively; the VH 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: 246, 247, and 248, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 48, 95, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 254, and 255, respectively; 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: and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 258, 259, and 260, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 116, 117, and 118, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 265, 266, and 267, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 129, 130, and 131, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 273, and 274, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 132, 134, and 136, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 136, 138, and 139, respectively. and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 278, 279, and 280, 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: 286, 287, and 288, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 168, 169, and 170, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 278, 279, and 280, respectively. containing CDR-L1, CDR-L2, and CDR-L3 with NOs:294, 234, and 295, respectively;The VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 35, 181, and 182, respectively, 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: 299, 300, and 301, respectively. the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 197, 198, and 199, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 319, 320, and 321, respectively;

[0147] 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. the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 48, 95, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively. In some embodiments, the antibody or antigen-binding fragment thereof provided herein comprises a VH region comprising CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 35, 36, and 37, respectively, according to the Chothia numbering, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, 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 NOs:82, 83, and 84, respectively, according to the Chothia numbering, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs:246, 247, and 248, respectively.

[0148] 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, according to the Kabat numbering, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 233, 234, and 235, respectively. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VH region comprising CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 87, 88, and 84, respectively, according to the Kabat numbering, and a VL region comprising CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 247, and 248, respectively.

[0149] 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: 63, 64, and 60, respectively. and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 221, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 63, 64, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 221, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 75, 76, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 229, 222, and 223, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 75, 76, and 60, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 229, 222, and 223, 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: 246, 247, and 248, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 98, 99, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 254, and 255, respectively; 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: 108, 109, and 105, respectively. and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 258, 259, and 260, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 121, 122, and 118, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 265, 266, and 267, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 134, 135, and 131, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 246, 273, and 274, respectively; VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 132, 133, and 134, respectively, and VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 136, 137, and 138, respectively. and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 278, 279, and 280, 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: 286, 287, and 288, respectively; the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 173, 174, and 170, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 286, 287, and 288, respectively. containing CDR-L1, CDR-L2, and CDR-L3 with NOs:294, 234, and 295, respectively;The VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 185, 186, and 182, respectively, 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: 306, 234, and 307, respectively. the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs: 202, 203, and 199, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs: 319, 320, and 321, respectively;

[0150] 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:241, 234, and 242, respectively. the VH comprises CDR-H1, CDR-H2, and CDR-H3 comprising SEQ ID NOs:98, 99, and 84, respectively, and the VL comprises CDR-L1, CDR-L2, and CDR-L3 comprising SEQ ID NOs:246, 247, and 248, respectively.

[0151] 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:3 and 19, 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:8 and 25, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs:9 and 26, respectively; the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs:10 and 27, respectively; and the VH and VL regions of the antibody or antigen-binding fragment thereof comprise the sequences of SEQ ID NOs:11 and 28, respectively.

[0152] 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; and 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.

[0153] In some embodiments, the antibody or antigen-binding fragment comprises one or more VHs and one or more VLs 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 linked directly or indirectly, optionally via a linker.

[0154] In some embodiments, an antibody or antigen-binding fragment, e.g., an scFv, can include 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., an scFv, can include a VL region or a portion thereof, followed by a linker, followed by a VH region or a portion thereof.

[0155] In some embodiments, the antibody or antigen-binding fragment thereof 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 that includes only the VH region. In some embodiments, the antibody or antigen-binding fragment is an scFv that includes a heavy chain variable (VH) region and a light chain variable (VL) region. In some embodiments, the single-chain antibody fragment (e.g., scFv) includes one or more linkers that join two antibody domains or regions, such as the VH region and the VL region. The linker is typically a peptide linker, such as a flexible and / or soluble peptide linker. Some linkers include those that are rich in glycine and serine, and / or, in some cases, rich in threonine. In some embodiments, the linker further includes charged residues, such as lysine and / or glutamic acid, that can improve solubility. In some embodiments, the linker further includes one or more prolines.

[0156] Accordingly, the anti-PD-L1 antibodies provided include single chain antibody fragments, such as scFvs, and diabodies, particularly human single chain antibody fragments that typically include a linker joining two antibody domains or regions, such as the VH and VL regions. The linker is typically a peptide linker, such as a flexible and / or soluble peptide linker, such as one that is rich in glycine and serine.

[0157] The antibody or antigen-binding fragment thereof may contain at least a portion of an immunoglobulin constant region, e.g., one or more constant region domains. In some embodiments, the constant region comprises a light chain constant region and / or a heavy chain constant region 1 (CH1). In some embodiments, the antibody comprises a CH2 domain and / or a CH3 domain, e.g., an Fc region. In some embodiments, the Fc region is an Fc region of a human IgG, e.g., an IgG1 or IgG4.

[0158] Nucleic acids, e.g., polynucleotides, encoding antibodies and / or portions thereof, e.g., strands, are also provided. Among the nucleic acids provided are those that encode the anti-PD-L1 antibodies and fragments described herein. Nucleic acids, e.g., polynucleotides, encoding one or more antibodies and / or portions thereof, e.g., one or more of the anti-PD-L1 antibodies or antigen-binding fragments described herein, and / or other antibodies and / or portions thereof, e.g., antibodies and / or portions thereof that bind other target antigens, are also provided. Nucleic acids can include those that include naturally occurring nucleotides and bases and / or non-naturally occurring nucleotides and bases, e.g., those with backbone modifications. The terms "nucleic acid molecule," "nucleic acid," and "polynucleotide" can be used interchangeably and refer to a polymer of nucleotides. Such polymers of nucleotides may contain naturally occurring nucleotides and / or non-naturally occurring nucleotides, including, but not limited to, DNA, RNA, and PNA. A "nucleic acid sequence" refers to the linear sequence of nucleotides that make up a nucleic acid molecule or polynucleotide.

[0159] Also provided are vectors containing nucleic acids, e.g., polynucleotides, for producing antibodies or antigen-binding fragments thereof, and host cells containing these vectors. Also provided are methods for producing antibodies or antigen-binding fragments thereof. The nucleic acids may encode an amino acid sequence comprising an antibody VL region, and / or an amino acid sequence comprising a VH region (e.g., an antibody light chain and / or a heavy chain). The nucleic acids may encode one or more antibody VL regions, and / or an amino acid sequence comprising a VH region (e.g., an antibody light chain and / or a heavy chain). In some embodiments, the nucleic acids, e.g., polynucleotides, encode one or more antibody VH regions and / or one or more VL regions in any order or orientation. In some embodiments, the nucleic acids, e.g., polynucleotides, encode the VH region and the VL region, and the coding sequence for the VH region is upstream of the coding sequence for the VL region. In some embodiments, the nucleic acids, e.g., polynucleotides, encode the VH region and the VL region, and the coding sequence for the VL region is upstream of the coding sequence for the VH region.

[0160] In some embodiments, one or more vectors (e.g., expression vectors) comprising such nucleic acids are provided. In further embodiments, host cells comprising such nucleic acids are provided. In some embodiments, the host cell comprises (e.g., is transformed by) (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising a VL region of an antibody and an amino acid sequence comprising a VH region of an antibody, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising a VL region of an antibody, and a second vector comprising a nucleic acid encoding an amino acid sequence comprising a VH region of an antibody. In some embodiments, the host cell comprises (e.g., is transformed by) 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, e.g., 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, one or more host cells may express different antibodies or may express the same antibody. In some embodiments, each of the host cells may express multiple antibodies.

[0161] Methods of making anti-PD-L1 antibodies (e.g., antigen-binding fragments) are also provided. For recombinant production of anti-PD-L1 antibodies, for example, a nucleic acid sequence or polynucleotide encoding the antibody, as described above, can be isolated and inserted into one or more vectors for further cloning and / or expression in a host cell. Such nucleic acid sequences can be readily isolated and sequenced using conventional techniques (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the antibody heavy and light chains). In some embodiments, methods of making anti-PD-L1 antibodies are provided, comprising culturing a host cell containing a nucleic acid sequence encoding the antibody provided above under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0162] In addition to prokaryotes, eukaryotic microbes, such as filamentous fungi or yeast, for example, fungal and yeast strains in which the glycosylation pathway has been modified to mimic or approximate that of human cells, resulting in the production of antibodies with partially or fully human glycosylation patterns, are also suitable hosts for cloning or expression of antibody-encoding vectors. See Gerngross, Nat. Biotech. 22:1409-1414 (2004) and Li et al., Nat. Biotech. 24:210-215 (2006).

[0163] Exemplary eukaryotic cells that can be used to express polypeptides include, but are not limited to, COS cells, e.g., COS7 cells; 293 cells, e.g., 293-6E cells; CHO cells, e.g., CHO-S cells, DG44 cells, Lec13 CHO cells, and FUT8 CHO cells; PER.C6® cells; and NSO cells. In some embodiments, the heavy and / or light chains (e.g., VH and / or VL regions) of the antibody can be expressed in yeast. See, e.g., U.S. Publication US 2006 / 0270045 A1. In some embodiments, a particular eukaryotic host cell is selected based on its ability to provide the desired post-translational modifications to the heavy and / or light chains (e.g., VH and / or VL regions). For example, in some embodiments, CHO cells produce polypeptides with higher levels of sialylation than the same polypeptides produced in 293 cells.

[0164] In some embodiments, the antibody or antigen-binding fragment provided herein is 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).

[0165] The embodiments provided further include vectors and host cells and other expression systems for the expression and production of antibodies and other antigen binding proteins, e.g., eukaryotic and prokaryotic host cells, such as bacteria, filamentous fungi, and yeast, as well as mammalian cells, e.g., human cells, and cell-free expression systems.

[0166] 2. Exemplary Features of Anti-PD-L1 Antibodies In some aspects, the antibodies or antigen-binding fragments thereof that are provided have one or more specified functional characteristics, eg, binding properties, eg, binding to a particular epitope.

[0167] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to PD-L1 protein. In some embodiments provided herein, PD-L1 protein refers to human PD-L1, non-human primate (e.g., cynomolgus monkey) PD-L1 protein, or mouse PD-L1 protein. In some embodiments provided herein, PD-L1 protein refers to human PD-L1 or non-human primate (e.g., cynomolgus monkey) PD-L1 protein. In some embodiments of any of the embodiments herein, PD-L1 protein refers to human PD-L1 protein. The observation that an antibody or other binding molecule binds to or specifically binds to PD-L1 protein does not necessarily mean that it binds to PD-L1 protein of any species. For example, in some embodiments, a binding characteristic with PD-L1 protein, e.g., the ability to bind specifically and / or to bind to a particular degree with a particular affinity, may in some embodiments refer to the ability with respect to human PD-L1 protein, and the antibody may not have this characteristic with respect to the PD-L1 protein of other species, e.g., a mouse.

[0168] In some embodiments, the antibody or antigen-binding fragment binds to a mammalian PD-L1 protein, including naturally occurring variants of PD-L1, such as certain splice variants or allelic variants.

[0169] In some embodiments, the antibody specifically binds to an extracellular domain of human PD-L1 protein, e.g., an epitope or region of human PD-L1 protein, e.g., the human PD-L1 extracellular domain 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 monkey PD-L1 protein, e.g., the cynomolgus monkey PD-L1 extracellular domain 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 also binds at a lower level or with a lower degree or affinity to cynomolgus monkey PD-L1 protein, such as the cynomolgus monkey PD-L1 extracellular domain 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, or binds at a lower level or with a lower degree or affinity to mouse PD-L1 protein, such as the mouse PD-L1 extracellular domain set forth in SEQ ID NO:329 (amino acid residues 19-238 of UniProt Accession No. Q9EP73). In some embodiments, the antibody binds to mouse PD-L1 protein with a lower affinity than it binds to human PD-L1 protein and / or cynomolgus monkey PD-L1 protein. In some embodiments, the antibody binds to mouse PD-L1 protein and / or cynomolgus monkey PD-L1 protein with a lower affinity than it binds to human PD-L1 protein, hi some embodiments, the antibody binds to cynomolgus monkey PD-L1 protein with a similar binding affinity compared to its binding to human PD-L1 protein.

[0170] 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 10% or about 10% of the antibody's binding to human PD-L1 protein, e.g., as measured by radioimmunoassay (RIA), ELISA, or surface plasmon resonance (SPR).In some embodiments, included among the antibodies provided are antibodies that bind to mouse PD-L1 protein at less than 10%, or at 10% or about 10% of the antibody's binding to human PD-L1 protein. In some embodiments, among the antibodies provided are antibodies that bind to cyno PD-L1 protein that is less than, or about, 50%, 40%, 30%, 20%, or 10% of the antibody's binding to human PD-L1 protein.

[0171] In some embodiments, the antibodies provided can bind to a PD-L1 protein, e.g., a human PD-L1 protein, with at least some affinity, as measured by any of a number of known methods. In some embodiments, affinity is measured by the equilibrium dissociation constant (K D ) and in some embodiments, affinity is represented by EC 50 It is represented by:

[0172] 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, e.g., a PD-L1 protein). For example, it is within the level of ordinary skill in the art to determine the binding affinity of a binding molecule, e.g., an antibody, to an antigen, e.g., PD-L1, e.g., human PD-L1 or cynomolgus PD-L1 or mouse PD-L1, by using any of a number 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. Patent Nos. 5,283,173, 5,468,614, or equivalents thereof).

[0173] For example, in some embodiments, a BIAcore® instrument may be used to determine the binding kinetics and constants of a complex between two proteins (e.g., between an antibody, e.g., an anti-PD-L1 antibody or fragment thereof, and an antigen, e.g., PD-L1 protein) using surface plasmon resonance (SPR) analysis. SPR measures the change in concentration of molecules at a sensor surface as the molecules bind to or dissociate from the surface. The change in SPR signal is directly proportional to the change in mass concentration near the surface, thus allowing the measurement of the binding kinetics between the two molecules. The dissociation constant for the complex may be determined by monitoring the change in refractive index over time as a buffer solution is passed over the chip.

[0174] Other suitable assays for measuring binding of one protein to another include, for example, immunoassays, such as enzyme-linked immunosorbent assays (ELISAs) and radioimmunoassays (RIAs), or determining binding by monitoring changes in the spectroscopic or optical properties of the proteins by fluorescence, UV absorption, circular dichroism, or nuclear magnetic resonance (NMR). Other exemplary assays include, but are not limited to, Western blots, ultracentrifugation, spectroscopy, flow cytometry, and other methods for detecting protein binding.

[0175] In some embodiments, the binding affinity of the provided antibodies, fragments, or conjugates to a PD-L1 protein, e.g., human PD-L1 protein, is similar to or higher than a reference antibody (e.g., avelumab). In some embodiments, the binding affinity is measured using the 50% effective concentration (EC 50 ) or dissociation constant (K D ) is measured as EC 50 or K D can be determined for binding to free or immobilized PD-L1, e.g., the extracellular domain of PD-L1, or PD-L1 expressed on the surface of a cell. In some embodiments, the EC of a provided antibody, fragment, or conjugate to a PD-L1 protein, e.g., a human PD-L1 protein, is determined for binding to the PD-L1 protein. 50 or K D The value of is similar to or lower (e.g., corresponding to a higher binding affinity) than a reference antibody (e.g., avelumab).

[0176] In some embodiments, binding affinity is measured using the 50% effective concentration (EC 50 In some embodiments, the EC50 value for binding of the antibody, fragment, or conjugate to a PD-L1 protein, e.g., a human PD-L1 protein, is measured as the EC50 value for binding of the antibody, fragment, or conjugate to a PD-L1 protein, e.g., a human PD-L1 protein. 50is 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 is similar to or lower than 50 is decreased by about 0.5 times, about 1 times, about 1.5 times, about 2 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.

[0177] In some embodiments, the binding affinity is determined by the K D In some embodiments, the K for binding of the antibody, fragment, or conjugate to a PD-L1 protein, e.g., a human PD-L1 protein, is measured as D is 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 similar to or lower than D is decreased by 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.

[0178] In some embodiments, the binding affinities of binding molecules, e.g., anti-PD-L1 antibodies, to different antigens, e.g., PD-L1 proteins from different species, can be compared to determine cross-species cross-reactivity. For example, cross-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, e.g., PD-L1 proteins from different species, e.g., human, cynomolgus monkey, or mouse, can be compared to determine cross-species cross-reactivity. D or EC 50 can be compared to determine cross-species cross-reactivity. In some embodiments, the cross-species cross-reactivity of an anti-PD-L1 antibody may be high, e.g., the anti-PD-L1 antibody binds to human PD-L1 and heterologous variant PD-L1 to a similar extent, e.g., the EC 50 or K D and EC against heterologous variant PD-L1 50 or K D is 1 or about 1. In some embodiments, the cross-species cross-reactivity of an anti-PD-L1 antibody may be low, for example, an anti-PD-L1 antibody has high affinity for human PD-L1, but low affinity for xenovariant PD-L1, or vice versa. The degree of cross-species cross-reactivity may be compared to the cross-species cross-reactivity of known antibodies.

[0179] In some embodiments, the provided antibodies or antigen-binding fragments thereof bind to human PD-L1 protein and non-human PD-L1 protein or other non-PD-L1 proteins to a similar extent. For example, in some embodiments, the provided antibodies or antigen-binding fragments thereof bind to a certain EC with human PD-L1 protein, e.g., a human PD-L1 protein 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. 50 or K Dand a non-human PD-L1, such as cynomolgus PD-L1, such as the cynomolgus PD-L1 protein set forth in SEQ ID NO:328 (amino acid residues 19-239 of UniProt Accession No. G7PSE7), has a similar or nearly identical EC 50 or K D In other embodiments, the provided antibodies or antigen-binding fragments thereof have an EC ratio (EC) to non-human PD-L1, e.g., mouse PD-L1, e.g., the mouse PD-L1 protein set forth in SEQ ID NO:329 (amino acid residues 19-238 of UniProt Accession No. Q9EP73). 50 or K D Lower EC 50 or K D and binds to human PD-L1 protein, for example, the human PD-L1 protein comprising the amino acid sequence of SEQ ID NO:327 (amino acid residues 19 to 239 of UniProt Accession No. Q9NZQ7), or an allelic variant or splice variant thereof.

[0180] In some embodiments, the antibodies (e.g., antigen-binding fragments) show binding preference to PD-L1 expressing cells compared to PD-L1 negative cells, e.g., certain cells known to express PD-L1 and / or described herein as expressing PD-L1, and certain cells known not to express PD-L1. In some embodiments, binding preference is observed when a significantly higher degree of binding to PD-L1 expressing cells compared to non-expressing cells is measured. In some embodiments, the binding preference is observed when a significantly higher degree of binding to PD-L1 expressing cells is measured, e.g., as measured by the mean fluorescence intensity and / or dissociation constant or EC 50The fold change in the extent of detected binding to PD-L1 expressing cells compared to non-PD-L1 expressing cells, as measured by, is at least, or at least about, 1.5, 2, 3, 4, 5, 6, or more, and / or is approximately the same, approximately the same, at least the same, or at least approximately the same, or more, as the fold change observed for the corresponding form of the reference antibody.

[0181] The anti-PD-L1 antibodies (e.g., antigen-binding fragments) provided herein can be identified, screened, or characterized for physical / chemical properties and / or biological activity by a variety of known assays. In one aspect, the antibodies can be tested for antigen-binding activity by, for example, known methods, such as ELISA, Western blotting, and / or flow cytometry assays, such as cell-based binding assays that assess binding of an antibody (e.g., conjugated or tagged with a fluorescent marker) to cells expressing a target antigen, e.g., PD-L1, in some cases compared to results using cells that do not express the target antigen, e.g., PD-L1. Binding affinity is expressed by the K D or EC 50 In some instances, the binding affinity, binding kinetics, and / or binding constant can be measured using an assay for determining molecular interactions, such as surface plasmon resonance analysis.

[0182] In some aspects, the anti-PD-L1 antibodies (e.g., antigen-binding fragments) provided herein may be conjugated to one or more agents, e.g., one or more drugs, small molecules, dyes (e.g., phthalocyanine dyes, e.g., IR700), and / or therapeutic agents, e.g., as described in Section II.

[0183] In some aspects, after conjugation with one or more agents, a conjugate comprising a provided antibody or antigen-binding fragment retains or substantially retains physical / chemical properties and / or biological activity such that the physical / chemical properties and / or biological activity of the conjugate are equivalent or similar to, or 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 are 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 physical / chemical properties and / or biological activity of the unconjugated (naked) antibody or antigen-binding fragment.

[0184] In some aspects, after conjugation with one or more agents, such as a phthalocyanine dye, such as IR700, a conjugate comprising a provided antibody or antigen-binding fragment retains or substantially retains binding affinity to the target antigen PD-L1, such that the binding affinity of the conjugate is equal to or similar to that 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 PD-L1 protein compared to the unconjugated antibody.

[0185] Competition assays may be used to identify antibodies that compete with any of the antibodies (e.g., antigen-binding fragments) described herein, or other known anti-PD-L1 antibodies. Assays to map the epitopes to which antibodies and other known anti-PD-L1 antibodies bind may also be used and are known.

[0186] B. Multispecific antibodies In certain embodiments, the PD-L1 binding molecule, e.g., an antibody or polypeptide, e.g., a chimeric receptor containing it, is multispecific. Among the multispecific binding molecules are multispecific antibodies, e.g., bispecific antibodies or trispecific antibodies. A multispecific binding partner, e.g., an antibody, has binding specificities for at least two different sites, which may be in the same antigen or in different antigens. In certain embodiments, one of the binding specificities is for PD-L1 and the other is for another antigen. In certain embodiments, the 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, the bispecific antibody may bind to two different epitopes of PD-L1. Bispecific antibodies may also be used to localize fluorescent, phototoxic, and / or cytotoxic agents to cells expressing PD-L1. Bispecific antibodies may be prepared as full length antibodies or antibody fragments. Among the multispecific antibodies are included multispecific single chain antibodies, such as diabodies, triabodies and tetrabodies, tandem di-scFvs, and tandem tri-scFvs.

[0187] In some embodiments, the second or additional antigens for the multiple targeting strategy include those in which at least one of the antigens is a universal tumor antigen or a family member thereof. In some embodiments, the second or additional antigen is an antigen expressed in tumors. In some embodiments, the PD-L1 binding molecules provided herein target an antigen of the same tumor type as the second or additional antigen. In some embodiments, the second or additional antigen may be a universal tumor antigen or a tumor antigen specific to a tumor type.

[0188] 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 factors, VEGF, PIGF, SIRPα, ED-B fibronectin, oncogenes, oncogene products, CD66a-d, necrosis antigens, and the like. 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, melanoma preferentially expressed antigen (PRAME), survivin, EGP2, EGP40, TAG72, B7-H6, IL-13 receptor a2 (IL-13Ra2), CA9, CD171, G250 / CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7 / 8, avb6 integrin, 8H9, NCAM, VEGF receptor, 5T4, fetal AchR, NKG2D ligand, dual antigen, universal tag-associated antigen, cancer-testis antigen, MUC1, MUC16, NY-ESO-1, MART-1, gp100, oncofetal antigen, VEGF-R2, carcinoembryonic antigenantigen (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, CD123, L1 cell adhesion molecule, MAGE-A1, MAGE A3, cyclins, 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, neoepitopes or neoantigens thereof. In some embodiments, the antigen is associated with or is a universal tag.

[0189] In some aspects, the antigen, e.g., the second or additional antigen, e.g., a disease-specific antigen and / or disease-associated antigen, is one expressed in multiple myeloma, e.g., G protein-coupled receptor class C group 5 member D (GPRC5D), CD38 (cyclic ADP ribose hydrolase), 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 those present in lymphoid tumors, myeloma, AIDS-related lymphoma, and / or post-transplant lymphoproliferation, such as CD38.Antibodies or antigen-binding fragments against such antigens are known, and include those described in, for example, U.S. Patent No. 8,153,765; No. 8,603,477, No. 8,008,450; U.S. Publication No. US20120189622 or US20100260748; and / or International PCT Publication No. 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.

[0190] C. Variants In certain embodiments, the antibody (e.g., antigen-binding fragment) comprises one or more amino acid variations, e.g., substitutions, deletions, insertions, and / or mutations, compared to the antibody sequences described herein. Exemplary variants include those 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 made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen binding.

[0191] 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 sequences of natural repertoires, e.g., human repertoires. Sites of interest for substitution mutagenesis include CDRs and FRs. Amino acid substitutions can be introduced into the antibody of interest, and the product screened for the ability to promote desired activity, e.g., retained / improved antigen binding, reduced immunogenicity, improved half-life, and / or improved effector function, e.g., antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC).

[0192] In some embodiments, one or more residues are substituted in the CDRs of a parent antibody (e.g., a humanized or human antibody). In some embodiments, the substitutions are made to, e.g., to reduce the possibility of immunogenicity upon administration to a human subject, e.g., to revert a sequence or position within a sequence to a germline sequence, e.g., an antibody sequence found in the germline (e.g., human germline).

[0193] In certain embodiments, substitution, insertion, or deletion can be made in 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 (e.g., conservative substitutions provided herein) that do not substantially reduce binding affinity can be made in the CDRs.Such changes can be, for example, other than antigen contact residues in the CDRs.In certain embodiments of the sequences of the VH and VL regions of the variants provided above, each CDR is unchanged or contains no more than one, two, or three amino acid substitutions.

[0194] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues. An example of a terminal insertion includes an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include the fusion to the N- or C-terminus of the antibody of an enzyme or a polypeptide which increases the serum half-life of the antibody.

[0195] D. Qualification In certain embodiments, the antibody is altered to increase or decrease the extent to which the antibody is glycosylated, e.g., by altering the amino acid sequence, by removing or inserting one or more glycosylation sites, and / or by modifying the oligosaccharides attached to the glycosylation sites, e.g., using certain cell lines. In some embodiments, N-linked glycosylation, which is a glycosylation site present at an asparagine residue within the consensus sequence -Asn-Xaa-Ser / Thr, is removed or inserted.

[0196] Exemplary modifications, variants, and cell lines are described, for example, in patent publications 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 2005 / 035586; WO 2005 / 035778;WO2005 / 053742;WO2002 / 031140;Okazaki et al.J.Mol.Biol.336:1239-1249(2004);Yamane-Ohnuki et al.Biotech.Bioeng.87:614(2004);Ripka et al. al.Arch.Biochem.Biophys.249:533-545(1986);US 2003 / 0157108 A1, 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.).

[0197] Among the modified antibodies are those with one or more amino acid modifications in the Fc region, e.g., those with a human Fc region sequence or other portion of the constant region (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) that includes 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, to alter binding to one or more types of Fc receptors, and / or to alter effector function.

[0198] 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 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 the antibodies, antibody fragments, and conjugates described herein.

[0199] 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 function (e.g., ADCC, ADCP, and / or CDC). In some of such embodiments, the anti-PD-L1 antibody comprises an Fc receptor that contains an amino acid substitution to eliminate a glycosylation site at a position corresponding to position 297 of the heavy chain according to EU numbering as described by 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 at, or equivalent to, position 297 according to EU numbering of the antibody sequence (N297Q), an asparagine to alanine substitution at, or equivalent to, position 297 according to EU numbering of the antibody sequence (N297A), or an asparagine to glycine substitution at, or equivalent to position 297 according to EU numbering of the antibody sequence (N297G).

[0200] In some of any of the provided embodiments, the anti-PD-L1 antibodies comprise 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, hi some embodiments, the anti-PD-L1 antibodies exhibit 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 (K334A), or a selenium to alanine substitution at position 257 (K334A). alanine to leucine substitution at position 330 (A330L), isoleucine to glutamic acid substitution at position 333 (E333A), glutamic acid to alanine substitution at position 333 (E333A), lysine to alanine substitution at position 334 (K334), and / or alanine to glutamine substitution at position 378 (A378Q). In some embodiments, the Fc region contains a glycine to alanine substitution at position 236, a serine to aspartic acid substitution at position 239, and an isoleucine to glutamic acid at position 332 (G236A / S239D / I332E) according to EU numbering of the antibody heavy chain.In some embodiments, the Fc region contains 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) according to 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) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a serine to aspartic acid substitution at position 239 and an isoleucine to glutamic acid substitution at position 332 (S239D / I332E) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a serine to alanine substitution at position 298 and a lysine to alanine substitution at position 334 (S298A / K334) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a glutamic acid to alanine substitution at position 333 and a lysine to alanine substitution at position 334 (E333A / K334) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a serine to alanine substitution at position 298, a glutamic acid to alanine substitution at position 333 and a lysine to alanine substitution at position 334 (S298A / E333A / K334) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains an arginine to alanine substitution at position 255 and a serine to alanine substitution at position 267 (R255A / S267A) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a threonine to alanine substitution at position 256 (T256A) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains a lysine to alanine substitution at position 334 and an alanine to glutamine substitution at position 378 (K334 / A378) according to EU numbering of the antibody heavy chain.In some embodiments, the Fc region contains 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 (S239D / A330L / I332E) according to 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, and an isoleucine to glutamic acid substitution at position 332 (G236A / S239D / I332E) according to EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains an arginine to serine substitution at position 325 and a leucine to phenylalanine substitution at position 328 (N325S / L328F) according to 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) according to the EU numbering of the antibody heavy chain. In some embodiments, the Fc region contains 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) according to the EU numbering of the antibody heavy chain.In some embodiments, the Fc region comprises 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 methionine substitution at position 298, according to EU numbering in one of the antibody heavy chains. It contains an alanine substitution (L234Y / L235Q / G236W / S239M / H268D / D270E / S298A), as well as, according to EU numbering on the opposing heavy chain, an aspartic acid to glutamic acid substitution at position 270, a lysine to aspartic acid substitution at position 326, an alanine to methionine substitution at position 330, and a lysine to glutamic acid substitution at position 334 (D270E / K326D / A330M / K334E).

[0201] Among the variants are cysteine ​​engineered antibodies, e.g., "ThioMAbs," and other cysteine ​​engineered variants, in which one or more residues of the antibody are substituted with cysteine ​​residues to generate reactive thiol groups at accessible sites, e.g., for use in drug and linker-drug conjugation, e.g., to create immunoconjugates. Cysteine ​​engineered antibodies are described, e.g., in U.S. Patent Nos. 7,855,275 and 7,521,541.

[0202] In some embodiments, the antibody (e.g., antigen-binding fragment) is modified to contain additional non-proteinaceous moieties, e.g., water-soluble polymers. Exemplary polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (either homopolymers or random copolymers), and dextran or poly(n-vinylpyrrolidone) polyethylene glycol, polypropylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, 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 be of any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if multiple polymers are attached, they may be the same molecule 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 in therapy for a defined condition, etc.

[0203] In some embodiments, the polynucleotide has been optimized, or contains certain features designed for optimization, e.g., for codon usage and / or to modify, e.g., increase or make more consistent expression, e.g., surface expression, of the encoded antibody or fragment between cell product lots. In some embodiments, a polynucleotide encoding an anti-PD-L1 antibody has been modified to remove cryptic or hidden splice sites.

[0204] II. Conjugates In some embodiments, the anti-PD-L1 antibody or antibody fragment (e.g., 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 a specific embodiment, 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.

[0205] A. Cytotoxic substances Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, and radioisotopes of Lu); chemotherapeutic agents (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other intercalating agents); growth inhibitors; enzymes and fragments thereof, e.g., nucleases; antibiotics; toxins, e.g., small molecule toxins, or toxins having enzymatic activity. In some embodiments, the antibody is conjugated to one or more cytotoxic agents, e.g., chemotherapeutic agents or drugs, growth inhibitors, toxins (e.g., protein toxins, toxins having enzymatic activity, derived from bacteria, fungi, plants, or animals), or radioisotopes.

[0206] Among the immunoconjugates are antibody-drug conjugates (ADCs) in which an antibody is conjugated to one or more drugs, including, but not limited to, maytansinoids; auristatins, such as monomethylauristatin drug moieties DE and DF (MMAE and MMAF); dolastatins; calicheamicin or derivatives thereof; anthracyclines, such as daunomycin or doxorubicin; methotrexate; vindesine; taxanes, such as docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; trichothecenes; and CC1065.

[0207] In some immunoconjugates, the antibody (e.g., antigen-binding fragment) is an antibody that is capable of binding 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, α-sarcin, Aleurites fordii protein, dianthin protein, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitors, gelonin, mitogellin, restrictocin, phenomycin, enomycin, as well as those conjugated to enzymatically active toxins or fragments thereof, including, but not limited to, the trichothecenes.

[0208] Non-limiting exemplary cytotoxic agents include aplidine, azaribine, anastrozole, azacitidine, bleomycin, bortezomib, bryostatin-1, busulfan, calicheamicin, camptothecin, 10-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin, irinotecan (CPT-11), SN-38, carboplatin, cladribine, cyclophosphatase, and the like. , cytarabine, dacarbazine, docetaxel, dactinomycin, daunomycin glucuronide, daunorubicin, 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, flutamine Fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone caproate, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, leucovorin, lomustine, mechlorethamine, medroprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, phenylbutyrate, prednisone , procarbazine, paclitaxel, pentostatin, PSI-341, semustine, streptozocin, tamoxifen, taxane, taxol, 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.

[0209] Among the immunoconjugates are those in which an antibody (e.g., an antigen-binding fragment) is conjugated with 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 , a radioactive isotope of Lu.

[0210] B. Phototoxic substances In certain aspects, the anti-PD-L1 antibody or antibody fragment (e.g., antigen-binding fragment) is conjugated to a phototoxic agent. In some aspects, the phototoxic agent is a phthalocyanine dye. Phthalocyanines are a group of photosensitive compounds that have a phthalocyanine ring system. Phthalocyanines are azaporphyrins (i.e., C32H18N8) that contain four benzoindole groups connected by nitrogen bridges in a 16-membered ring of alternating carbon and nitrogen atoms that form stable chelates with metal or metalloid cations. In these compounds, the center of the ring is occupied by a metal ion (either a diamagnetic or paramagnetic ion), which may bear zero, one, or two ligands, depending on the ion. Furthermore, the ring periphery can be either unsubstituted or substituted.

[0211] In some embodiments, phthalocyanines strongly absorb red or near infrared light, with an absorption peak at about 600 nm to 810 nm, which in some cases allows light to penetrate deep into tissue.Phthalocyanines are generally photostable.This photostability is typically advantageous in pigments and dyes, as well as many of the other applications of phthalocyanines.

[0212] In some embodiments, the phthalocyanine dye is water-soluble and contains a luminescent fluorophore moiety with at least one aqueous solubilizing moiety. In some embodiments, the aqueous solubilizing moiety contains silicon. In some embodiments, the phthalocyanine dye has a core atom, for example, Si, Ge, Sn, Al, or Zn. In some embodiments, the phthalocyanine dye contains a linker with a reactive group that can form a bond between the linker and another molecule, i.e., form a conjugate. In some embodiments, the phthalocyanine dye can be adjusted to fluoresce at a specific wavelength.

[0213] In some embodiments, the phthalocyanine dye contains a linker, ie, a linker-phthalocyanine dye moiety (LD).

[0214] In some embodiments, the phthalocyanine dye is a phthalocyanine dye having a silicon coordinated metal (Si-phthalocyanine dye). TIFF2025508739000004.tif85128 or a salt, stereoisomer, or tautomer thereof, wherein L is a linker; Q is a reactive group for attachment of the dye to a targeting molecule; R 2 , R 3 , R 7 , and R 8 are each independently selected from optionally substituted alkyl and optionally substituted aryl; 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, where R 4 , R5 , R 6 , R 9 , R 10 , and R 11 at least one of which contains a water-soluble group; 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 among hydrogen, halogen, optionally substituted alkylthio, optionally substituted alkylamino, and optionally substituted alkoxy; X 2 and X 3 each independently represents a C which may be interrupted by a heteroatom 1 ~C 10 It is alkylene.

[0215] In some embodiments, the phthalocyanine dye has the formula: TIFF2025508739000005.tif75137, its salts, stereoisomers, or tautomers, wherein X 1 and X 4 each independently represents a C which may be interrupted by a heteroatom 1 ~C 10 is alkylene; R 2 , R 3 , R 7 , and R 8 are each independently selected from optionally substituted alkyl and optionally substituted aryl; 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, where R 4 , R 5 , R 6 , R 9 , R 10 , and R 11 at least one of which contains a water-soluble group; R 16 , R 17 , R 18 , and R 19 are each independently selected from among hydrogen, halogen, optionally substituted alkylthio, optionally substituted alkylamino, and optionally substituted alkoxy.

[0216] In some embodiments of the methods and uses provided herein, the Si-phthalocyanine dye is IRDye 700DX (IR700). In some embodiments, the phthalocyanine dye containing a reactive group is IR700 NHS ester, such as IRDye 700DX NHS ester (LiCor 929-70010, 929-70011). In some embodiments, the dye has the following formula: TIFF2025508739000006.tif103155, or a salt, stereoisomer, or tautomer thereof.

[0217] For purposes herein, the term "IR700," "IRDye 700," or "IRDye 700DX" includes the above formula when the dye is conjugated, e.g., to an antibody, e.g., via a reactive group.

[0218] 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 the formula (X): TIFF2025508739000007.tif46128, or a salt, stereoisomer, or tautomer thereof, wherein M is a metal or metalloid; X is TIFF2025508739000008.tif22128; Y is TIFF2025508739000009.tif21128; R 1 and R 2 are each independently 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; R 3 , R 4 , or R 5 is selected from substituent (a) or substituent (b), (a)R 3 is hydrogen, -L 3 -H, -L 3 -A, or -L 3 -Z; 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; R 5 -L 5 -H or -L 5 -A; (b)R 3 -L 3 -H or -L 3 -A; R 4 -L 4-H, -(NH) m -L 4 -A, or -(O) m -L 4 -A; where R 3 and R 4 -L 4 -A is connected by a bond to form a substituted heterocyclyl; R 5 -L 5 -H or -L 5 -A; However, R 3 , R 4 , and R 5 at least one of is an A-containing group; A is a reactive group, or a protected or reacted form thereof, capable of forming a covalent bond with a thiol, hydroxyl, carboxyl, or amino group of a second moiety; R 6 and R 7 are each independently 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; R 8 , R 9 , or R 10 is selected from substituent (a) or substituent (b), (a)R 8 is hydrogen, -L 8 -H, or -L 8 -Z; R 9 -L 9 -H, -(NH) n -L 9 -Z, or -(O) n -L 9 -Z; R 10 -L 10 -Z; (b)R 8 and R 9 -L 9 -Z is connected by a bond to form a substituted heterocyclyl; R 10 -L 10 -H or -L 10 -Z; However, R 8 , R 9 , and R 10 at least one of is a Z-containing group; Z is a water-soluble group optionally substituted with A or L'-A; L 1 and L 2 are each independently an optionally substituted alkylene, an optionally substituted heteroalkylene, an optionally substituted alkenylene, or an optionally substituted heteroalkenylene, an optionally substituted cycloalkyl, or an optionally substituted heterocyclyl; L 3 , L 4 , L 5 , L 8 , L 9 , and L 10 are each independently an optionally substituted alkylene, an optionally substituted heteroalkylene, an optionally substituted alkenylene, an optionally substituted heteroalkenylene, an optionally substituted cycloalkylene, an optionally substituted heterocyclene, an optionally substituted arylene, an optionally substituted aralkylene, an optionally substituted heteroaralkylene, or an optionally substituted heteroarylene, wherein a carbon atom of the alkylene, heteroalkylene, alkenylene, heteroalkenylene, cycloalkylene, heterocyclene, arylene, aralkylene, heteroaralkylene, or optionally substituted heteroarylene is optionally further substituted with Z, and each nitrogen atom of the heteroalkylene or heteroalkenylene is optionally substituted with one or two L'-Z; each L' is independently an optionally substituted alkylene, an optionally substituted heteroalkylene, an optionally substituted alkenylene, an optionally substituted heteroalkenylene, an optionally substituted cycloalkylene, an optionally substituted heterocyclene, an optionally substituted arylene, an optionally substituted aralkylene, an optionally substituted heteroaralkylene, or an optionally substituted heteroarylene; a is 0 or 1; b is 0 or 1; c is 0 or 1; d is 0 or 1; m is 0 or 1; n is 0 or 1; However, if b is 1, then a is 0; If d is 1, then c is 0; If m is 1, then b is 1; If n is 1, then c is 1.

[0219] In certain embodiments, M is Si, Ge, Sn, or Al. In further certain embodiments, M is Si or Ge.

[0220] In certain embodiments, Provided herein is a conjugate containing a compound of formula (X) selected from the group consisting of: TIFF2025508739000010.tif207151TIFF2025508739000011.tif191134TIFF2025508739000012.tif207136, or a salt, stereoisomer, or tautomer thereof.

[0221] In a particular embodiment, the compound of formula (X) is TIFF2025508739000013.tif111128, or a salt, stereoisomer, or tautomer thereof.

[0222] In some embodiments, the phthalocyanine dye is selected from Table A.

[0223] (Table A) TIFF2025508739000014.tif93165TIFF2025508739000015.tif212165TIFF20255087390 00016.tif216165TIFF2025508739000017.tif213165TIFF2025508739000018.tif196165

[0224] In a specific embodiment, the phthalocyanine dye containing a reactive group has the formula (I): TIFF2025508739000019.tif73128, or a salt, stereoisomer, or tautomer thereof.

[0225] In a specific embodiment, the phthalocyanine dye containing a reactive group has the formula (II): TIFF2025508739000020.tif71128, or a salt, stereoisomer, or tautomer thereof.

[0226] C. Other Features In some embodiments, provided herein is a conjugate comprising a Si-phthalocyanine dye linked to an anti-PD-L1 antibody or fragment provided herein, and for use in the methods 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.

[0227] In some embodiments, the provided anti-PD-L1 antibodies or anti-PD-L1 antibody conjugates contain a functional Fc region. In some embodiments, the anti-PD-L1 antibodies or conjugates do not comprise a functional Fc region. A non-functional Fc region includes an absent or truncated Fc region, or an Fc region containing amino acid substitutions or deletions that reduce or eliminate an Fc-mediated activity, such as an effector function (e.g., ADCC activity or ADCP activity). In some embodiments, the Fc region has been modified to exhibit ADCC activity and / or ADCP activity, or has been modified to exhibit enhanced effector function, such as enhanced ADCC activity and / or ADCP activity. Modifications to the Fc region that result in enhanced ADCC activity and / or ADCP activity may be any known modification or may be empirically determined. Exemplary modifications include, but are not limited to, modifications to the Fc region described herein. In some embodiments, the antibody is made to be defucosylated.

[0228] In some aspects, the provided conjugates of an anti-PD-L1 antibody or antibody fragment and one or more phototoxic, cytotoxic, or fluorescent agents can be made using any of a number 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 or cleavable linker. The linker can be a "cleavable linker" or "releasable linker" that facilitates 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. Patent No. 5,208,020). In some embodiments, the releasable or cleavable linker is released or cleaved in the presence of one or more conditions or factors present in the tumor microenvironment (TME), such as matrix metalloproteinases (MMPs), hypoxic conditions, or acidic conditions.

[0229] III. Methods of Treatment with Anti-PD-L1 Antibodies and Conjugates, and Uses of Anti-PD-L1 Antibodies and Conjugates In some embodiments, the provided anti-PD-L1 antibodies, fragments, or conjugates are used in methods and uses of treatment of lesions or tumors. In some embodiments, the provided methods and uses include 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 include administering the conjugate and illuminating a target area with light of an appropriate wavelength for use with the phthalocyanine dye, such that the light excites the dye and results in the killing of cells expressing PD-L1 on their surface, e.g., as described herein. Any of such methods and uses result in the enhancement, activation, induction, elicitation, strengthening, or support of immune function, e.g., local and / or systemic immunity, reduction or elimination of lesions (e.g., tumors), reduction or inhibition of tumor growth, reduction, inhibition, or elimination of tumor cell metastasis, or any combination thereof.

[0230] However, in some aspects, the provided compositions, methods, and uses can further enhance, activate, induce, trigger, strengthen, or support immune function, e.g., local and / or systemic immunity, by inhibiting PD-1:PD-L1 interaction and / or killing and eliminating cells expressing PD-L1 on their surface. Thus, eliminating or killing cells expressing PD-L1 on their surface, in particular immune cells with immunosuppressive function, e.g., M2 tumor-associated macrophages (M2 TAMs), M1- or M2-type macrophages, tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs), can function to enhance, activate, induce, trigger, strengthen, or support immune function, e.g., local and / or systemic immunity, e.g., anti-tumor or anti-cancer immunity. Elimination or killing of cells expressing PD-L1 on their surface can be brought about by Fc-mediated effector function, delivery of a toxic payload to the PD-L1 expressing cells, and / or following illumination of the target area with light of an appropriate wavelength for use with a photoactivatable dye, such that the light excites the dye and results in killing of the cells.

[0231] In some aspects, the provided methods and uses can enhance, activate, induce, recruit, or support lymphocyte infiltration into tumors or lesions. In some embodiments, the provided methods and uses activate intratumoral innate responses, resulting in increased intratumoral dendritic cell activation (e.g., activated dendritic cells). In some aspects, the provided methods and uses activate adaptive immune responses, resulting in increased infiltration of CD8+ T cells. In some embodiments, the provided methods and uses lead to a decrease in the number of exhausted CD8+ T cells in tumors. In some embodiments, the provided methods and uses lead to increased intratumoral infiltration of newly primed CD8+ T cells. In some embodiments, the provided methods and uses can lead to improved therapeutic outcomes, for example, by selecting subjects for treatment with higher levels or numbers of non-exhausted effector cells, e.g., CD8+ T cells, and / or by improving the activity or response of non-exhausted effector cells.

[0232] In some aspects, the provided compositions, methods and uses can be applied to tumors that are resistant or refractory to checkpoint inhibitor immunotherapy. In some aspects, the provided compositions, methods and uses can be applied to tumors that are resistant or refractory to anti-PD-L1 immunotherapy. Tumors, such as solid tumors, can develop resistance to checkpoint inhibitor immunotherapy by several mechanisms, including but not limited to irreversible T cell exhaustion, insufficient T cell priming, tumor cell immune escape (immunoediting) that leads to the upregulation of compensatory inhibitory signals of T cells, creation of immunosuppressive tumor microenvironment, such as increased infiltration of Treg, MDSC, 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 provided compositions, methods and uses are useful in overcoming one or more checkpoint inhibitor resistance mechanisms utilized by some tumors.

[0233] Furthermore, administration of the antibody, cytotoxic conjugate, or photoactivatable conjugate and subsequent irradiation can also result in the direct killing of cancer cells expressing PD-L1, thereby inhibiting or reducing tumor growth. The PD-L1 antibody, cytotoxic conjugate, or photoactivatable conjugate, e.g., anti-PD-L1-phthalocyanine conjugate, can directly or indirectly affect and kill tumor cells, or cells present in the tumor or tumor microenvironment (also called tumor microenvironment; TME), e.g., tumor cells in a location different from the primary tumor, metastasized tumors, newly arising tumor cells, and / or tumors of a different type or cell surface antigen expression. Thus, the provided compositions, methods, and uses can provide effective treatment for tumor cells that do not express cell surface PD-L1, tumors, lesions, or cancers that have a low response or are substantially non-responsive, have failed, subsequently relapsed, are refractory, and / or are resistant to previous treatments, e.g., previous immunomodulatory treatments. In specific embodiments, the compositions, methods, and uses provided can treat tumors or lesions that are non-responsive, resistant, or refractory to anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy.

[0234] In some embodiments, the compositions, methods, and uses provided herein are also effective for treating tumors that are larger in size and exhibit greater immunosuppression than smaller tumors. Such tumors may be hyporesponsive or non-responsive to other treatments, such as treatments with immunomodulators, such as immune checkpoint inhibitors (e.g., anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy). In such cases, anti-PD-L1 photoimmunotherapy provided by administration of the anti-PD-L1 conjugates described herein and subsequent irradiation may, in some cases, be effective in inhibiting or substantially reducing the growth of larger tumors that are not effectively inhibited by other immunomodulator therapy and / or anti-cancer therapy. In some embodiments, the compositions, methods, and uses provided herein are effective for treating tumors that are larger in size and resistant to anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy.

[0235] In some aspects, the method and use of treating tumor or lesion in a subject is provided by activating immune cell response.Immune cell activation can be direct or indirect activation.In some aspects, the method and use of treating the subject who has low response, has been non-responsive, has been resistant, has been refractory, has failed to respond, or has relapsed after previous immunotherapy (e.g., anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy) for tumor or lesion is provided.

[0236] In some aspects, the method comprises administering to a subject having a tumor or lesion a conjugate comprising a phthalocyanine dye, e.g., IR700, linked to an anti-PD-L1 antibody described herein. ..., at a wavelength of at or about 600 nm to at or about 850 nm, at or about 850 nm, at a dose of 25 J / cm, which can lead to the killing of PD-L1 expressing cells, thereby inhibiting the growth of the tumor or lesion. 2 Or about 25J / cm2 From 400J / cm 2 Or about 400 J / cm 2 irradiating the target area in which the PD-L1 expressing cells, e.g., PD-L1 expressing immune cells, are located with a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length. In some aspects, the method may also lead to the killing of PD-L1 expressing cells, thereby increasing the number or activity of immune cells in the tumor or lesion and / or the microenvironment of the tumor or lesion.

[0237] In some embodiments, the therapeutic effect of the present invention is more than 25 J / cm at wavelengths at or about 600 nm to 850 nm in a subject having a tumor or lesion administered a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 Methods and uses are provided for treating a tumor or lesion in a subject by activating an immune cell response comprising irradiating a target area in which PD-L1 expressing immune cells are located with a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, wherein the method leads to the killing of the PD-L1 expressing cells, 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.

[0238] In some embodiments, a subject having a tumor or lesion that has had a low response or has been non-responsive to previous immunotherapy administered a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1 has a PD-L1 concentration of 25 J / cm at wavelengths at or about 600 nm to 850 nm at or about 850 nm. 2 Or about 25J / cm2 From 400J / cm 2 Or about 400 J / cm 2 Methods and uses are provided for treating a subject who has had a low response, was non-responsive, was resistant, was refractory, failed to respond, or has subsequently relapsed to a previous immunotherapy for a tumor or lesion, comprising irradiating a target area in which PD-L1 expressing immune cells are located with a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, wherein the method leads to the killing of the PD-L1 expressing cells, thereby increasing the number or activity of immune cells in the tumor and / or tumor microenvironment. In some of any of the embodiments, the PD-L1 expressing cells are immune cells.

[0239] In some embodiments, methods and uses are provided for enhancing response to anti-cancer drugs in a subject with a tumor or lesion. In some aspects, the methods include administering an anti-cancer drug to a subject with a tumor or lesion. In some aspects, administering to the subject a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1. In some aspects, administering to the subject a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1 at a wavelength of at or about 600 nm to at or about 850 nm at a dose of 25 J / cm2 leads to greater inhibition of tumor or lesion growth compared to inhibition by treatment with the anti-cancer drug alone. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 irradiating the target area where the PD-L1 expressing immune cells are located with a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length.

[0240] In some embodiments, administering to a subject a conjugate that includes a phthalocyanine dye linked to a targeting molecule that binds to PD-L1; and administering to a subject a PD-L1 concentration of 25 J / cm at a wavelength at or about 600 nm to 850 nm at or about 850 nm. 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 and a dose of from about 2 J / cm fiber length to about 500 J / cm fiber length, respectively, of irradiating a target area in which PD-L1 expressing immune cells are located, wherein the PD-L1 expressing immune cells are irradiated with ...

[0241] In some embodiments, there is provided a method and use of enhancing a response to an anti-cancer drug in a subject having a tumor or lesion, comprising administering to the subject the anti-cancer drug, wherein the subject is administered a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1; and administering to the subject a phthalocyanine dye having a wavelength of at or about 600 nm to at or about 850 nm of at least 25 J / cm 2 Or about 25J / cm 2 From 400J / cm 2 Or about 400 J / cm 2 and following treatment comprising irradiation of a target area in which PD-L1 expressing immune cells are located at a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, wherein the administration and treatment of an anti-cancer agent leads to greater inhibition of tumor or lesion growth compared to inhibition by the anti-cancer agent alone.

[0242] In some embodiments, methods and uses are provided for vaccinating or immunizing a subject to generate anti-cancer immune responses. In some aspects, vaccination or immunization of a subject to generate anti-cancer immune responses can inhibit the growth and / or reduce the size of a first tumor or lesion; can also delay or prevent the appearance, growth, or establishment of one or more second tumors or lesions, for example, located distally from the treated first tumor or lesion. In some aspects, the method comprises administering to the subject a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1. In some aspects, the method comprises irradiating a target area, which leads to an anti-cancer response selected from delaying or inhibiting the appearance or growth of a tumor in the subject, or the appearance or increase of T memory cells in the vicinity of the tumor.

[0243] In some embodiments, there is provided a method and use of vaccinating or immunizing a subject to generate an anti-cancer immune response comprising irradiating a target area in a subject administered a conjugate comprising a phthalocyanine dye linked to a targeting molecule that binds to PD-L1, wherein the method and use lead to an anti-cancer response selected from delaying or inhibiting the appearance or growth of a tumor in the subject, or the appearance or increase of T memory cells in the vicinity of the tumor.

[0244] In some embodiments, one or more of the steps of the method are repeated.In some embodiments, the administration of the conjugate is repeated one or more times, and optionally, the irradiation step is repeated after each repeated administration of the conjugate.In some embodiments, the method further comprises administering an additional therapeutic agent or anticancer treatment.

[0245] A. Methods for Stimulating or Enhancing Anti-Cancer Immune Responses In some aspects, the provided methods and uses utilizing compositions comprising anti-PD-L1 conjugates can result in an enhanced immune response, e.g., a systemic immune response and / or a local immune response, in a subject, which in turn can result in an enhanced response to a therapy or treatment of a tumor, lesion, or cancer. In some aspects, the methods and uses herein include administering an anti-PD-L1 conjugate to a subject, and irradiating a target area, e.g., a target area in which PD-L1 expressing cells are present, e.g., a tumor, near a tumor, a lymph node, near a lymph node, or the tumor microenvironment, following administration of the conjugate.

[0246] In some aspects, the provided embodiments can stimulate, enhance, activate, induce, induce, boost, strengthen, or support an immune response, e.g., a systemic immune response, in a subject with a tumor, a lesion, or a cancer. In some embodiments, the provided methods and uses result in an enhanced systemic immune response in a subject with a tumor, a lesion, or a cancer. "Systemic immune response" refers to the ability of a subject's immune system to respond systemically to an immunological challenge, e.g., associated with a tumor, a lesion, or a cancer. A systemic immune response can include a systemic response of the subject's adaptive and / or innate immune system. A systemic immune response can include an anti-tumor or anti-cancer response from the subject's adaptive and / or innate immune system. In some aspects, a systemic immune response includes an immune response that extends to different tissues, e.g., the bloodstream, lymph nodes, bone marrow, spleen, and / or the tumor microenvironment, and in some cases includes a coordinated response in tissues and organs, and various cells and factors of tissues and organs. In some embodiments, the provided embodiments can stimulate, enhance, activate, induce, induce, boost, strengthen, or support the anti-cancer or anti-tumor immune response of the subject's own immune system, e.g., the adaptive immune system and / or the innate immune system. In some aspects, the provided methods and uses can result in an enhanced innate immune response in the subject.

[0247] In some aspects, provided embodiments may also exhibit an abscopal effect. In some aspects, "abscopal effect" refers to a treatment effect in which tumors not directly treated or tumors distant from the site of localized treatment, e.g., distant or metastatic tumors, are also treated, e.g., tumor volume is reduced.

[0248] In some aspects, the provided embodiments may provide tumor immunity. In such aspects, the provided embodiments prevent or impede new tumor growth or metastasis. In some embodiments, the inhibition of tumor growth provided by the provided embodiments leads to a sustained anti-tumor response. In some embodiments, the inhibition of tumor growth provided by the provided embodiments leads to extended progression-free survival. In some embodiments, the inhibition of tumor growth provided by the provided embodiments leads to a reduced probability of recurrence and / or a reduced probability of metastasis. In some aspects, the provided embodiments may provide immunity against the same or different tumor types in the treated subject. In some aspects, the provided embodiments may inhibit the growth of tumors from different tumor lineages, i.e., different types of tumors that have developed or may develop in the treated subject.

[0249] In some aspects, the target area is an area that contains PD-L1 expressing cells. In some embodiments, the PD-L1 expressing cells are immune cells. In some of the optional embodiments, the method directs the killing of PD-L1 expressing cells, e.g., PD-L1 expressing immune cells. In some embodiments, the PD-L1 expressing immune cells are selected from the group consisting of monocytes, macrophages, dendritic cells (DCs), M2 tumor-associated macrophages (M2 TAMs), M1 or M2 macrophages, tolerogenic dendritic cells (tDCs), and myeloid-derived suppressor cells (MDSCs). In some embodiments, the PD-L1 expressing immune cells are located in the tumor, the tumor microenvironment, or lymph nodes.

[0250] In some aspects, the target area irradiated by the provided embodiments is a tumor, e.g., a primary tumor, a tumor, e.g., near a primary tumor, or a tumor microenvironment (TME). In some embodiments, the target area is near a tumor, or proximal 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.

[0251] In some embodiments, the target area is a lymph node or near a lymph node. In some embodiments, the target area is a lymph node or near a lymph node, e.g., containing PD-L1 expressing cells. In some embodiments, the target area is a lymph node. In some embodiments, the target area is near a lymph node.

[0252] In some aspects, the provided embodiments can stimulate or enhance a systemic response, e.g., a systemic immune response, to one or more primary tumors or lesions and / or one or more secondary tumors or lesions, e.g., metastatic tumors or lesions, or tumors or lesions of a different type.

[0253] In some aspects, provided embodiments stimulate or enhance a subject's immune response, e.g., a subject's anti-cancer immune response, in some cases by eliminating PD-L1 expressing immune cells, e.g., those that may have immunosuppressive function, e.g., monocytes, macrophages, e.g., 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 aspects, provided embodiments stimulate or enhance a subject's immune response, e.g., a systemic immune response and / or a local immune response, that targets a tumor, lesion, or cancer, by killing and eliminating PD-L1 expressing immunosuppressive cells, e.g., M2 TAMs, tDCs, or MDSCs. As exemplified herein, depletion of a subject's CD8+ T cells resulted in tumor growth similar to that in saline-administered controls, so inhibition of tumor growth following administration of a PD-L1-phthalocyanine dye conjugate and light irradiation requires the presence and / or activity of the subject's CD8+ T cells. In some aspects, immunosuppressive cells, e.g., M2 TAMs, tDCs, or MDSCs, e.g., those expressing PD-L1, inhibit the function and / or activity of the subject's immune cells, e.g., CD8+ T cells or natural killer (NK) cells. By killing and eliminating immunosuppressive cells, e.g., PD-L1-expressing immunosuppressive cells, e.g., M2 TAMs, tDCs, or MDSCs, the provided embodiments can stimulate and enhance the subject's immune response. As exemplified herein, such treatment according to the provided embodiments results in inhibition of the growth of one or more primary tumors, e.g., a complete response to treatment, as well as inhibition of the growth of one or more second tumors, e.g., a second tumor of the same or different type and / or origin, and / or a second tumor present at a different site, e.g., a site distal to the primary tumor or lesion.

[0254] In some aspects, inhibition of tumor or lesion growth and / or killing of PD-L1 expressing cells is dependent on the presence of CD8+ T cells. In some embodiments, prior to administration, the subject has a tumor or lesion with a low number or level of CD8+ T cell infiltration. In some embodiments, following administration and irradiation, the number, level, or activity of immune cells is increased in the tumor or tumor microenvironment. In some embodiments, following administration and irradiation, the number or level of CD8+ T cell infiltration in the tumor or lesion is increased. In some embodiments, following administration and irradiation, the number of memory T cells in the vicinity of the tumor is increased.

[0255] In some aspects, the stimulated or enhanced systemic immune response includes systemic CD8 + Increased numbers and / or activity of T effector cells, increased systemic T cell cytotoxicity against tumor cells as measured using CTL assays using cells derived from the spleen, peripheral blood, bone marrow, or lymph nodes, intratumoral CD8 + Increased number, activity, and / or priming of T effector cells, systemic CD8 +These include increased T cell activation, increased systemic dendritic cell activation, increased dendritic cell activation in primary or secondary (e.g., metastatic or new) tumors or lesions, increased intratumoral dendritic cell infiltration in primary or secondary (e.g., metastatic or new) tumors or lesions, increased new T cell priming in primary or secondary (e.g., metastatic or new) tumors or lesions, increased T cell diversity in primary or secondary (e.g., metastatic or new) tumors or lesions, decreased systemic regulatory T cells, decreased regulatory T cells in primary or secondary (e.g., metastatic or new) tumors or lesions, decreased systemic myeloid-derived suppressor cells, decreased intratumoral myeloid-derived suppressor cells in primary or secondary (e.g., metastatic or new) tumors or lesions, decreased tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs) in primary or secondary (e.g., metastatic or new) tumors or lesions, or any combination thereof. In some cases, the systemic response can be assessed by collecting blood, tissues, cells, or other bodily fluids from the subject and assessing the increase in proinflammatory cytokines, the increase or appearance of immune cell activation markers, and / or T cell diversity. In some aspects, the systemic response can be assessed by assaying cells directly or indirectly affected by the method. For example, cells can be collected from the subject at 4-28 days after treatment or any time after the step of irradiating the primary tumor in the subject.

[0256] In some aspects, the provided embodiments can stimulate, enhance, boost, strengthen, or support immune responses, e.g., local responses, e.g., local immune responses, in subjects with tumors, lesions, or cancer. In some embodiments, the provided methods and uses result in enhanced local responses in subjects with tumors, lesions, or cancer. "Local immune response" refers to immune responses in tissues or organs to immunological challenges, e.g., those associated with tumors, lesions, or cancer. Local immune responses can include adaptive and / or innate immune systems. In some aspects, local immunity includes immune responses that occur simultaneously in different tissues, e.g., bloodstream, lymph nodes, bone marrow, spleen, and / or tumor microenvironment.

[0257] In some aspects, the stimulated or enhanced local immune response includes intratumoral CD8 + T effector cells (e.g., CD3 + CD8 + Increase in number and / or activity of CD8 + Increased T effector cell activation, intratumoral dendritic (CD11c + ) Increased cell infiltration, intratumoral dendritic cell activation (e.g., CD11c + CD80 + and / or CD11c + CD40 + ), and intratumoral antigen-presenting dendritic cells (CD11b + CD103 + CD11c + ), priming of new T cells within the tumor (e.g., CD3 + CD8 + PD1 - cells), increased intratumoral T cell diversity, and intratumoral neutrophils (CD11b + Cy6C - / low Ly6G + cells), intratumoral macrophages (e.g., CD11b + F4 / 80 + cells), intratumoral regulatory T cells (Treg), intratumoral myeloid-derived suppressor cells (MDSCs; e.g., CD11b+ Ly6C + Ly6G - Depletion of tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs) in the tumor, depletion of exhausted T cells in the tumor, e.g., exhausted CD8+ T cells (e.g., PD-1 + CTLA-4 + CD3 + CD8 + In some aspects, the local immune response includes a reduction in the number and / or activity of cells (cells), or any combination thereof. In some aspects, a stimulated or enhanced local immune response is provided by any of the embodiments. In some aspects, the cell surface phenotype of cells, e.g., immune cells, indicative of a local or innate immune response, can be assessed by staining with a reagent, e.g., a labeled antibody, that can be used to detect the expression of a marker on the surface. In some aspects, the cell surface phenotype of cells, e.g., immune cells, indicative of a local or innate immune response, is detected using flow cytometry.

[0258] In some cases, the local response, e.g., the local immune response, can be assessed by taking blood, tissue or other samples from the subject and assessing for an increase in anti-immune cell types in the tumor or TME and / or assessing an increase or appearance of local immune activation markers. In some aspects, the local response, e.g., the local immune response, can be assessed by assaying cells directly or indirectly affected by the method. For example, cells can be collected from the subject at 4-28 days after treatment or any time after the step of irradiating the primary tumor in the subject.

[0259] In some aspects, the method and use also include administering an additional therapeutic agent, such as an immunomodulator, such as an immune checkpoint inhibitor. The immunomodulator can be administered before, simultaneously, or after the administration of the conjugate. In some aspects, the administration of the additional therapeutic agent, such as an immunomodulator, can also contribute to stimulating, enhancing, activating, inducing, strengthening, or supporting an immune response, such as a subject's systemic immune response and / or local immune response, such as an anti-cancer response or anti-tumor response. Exemplary additional therapeutic agents, compositions, combinations, methods, and uses include those described herein, for example, in Section V.

[0260] B. Tumors and Lesions for Anti-PD-L1 Conjugate Treatment The methods described herein include administration of an anti-PD-L1 conjugate to achieve cell killing, e.g., of cells expressing PD-L1 on their surface, and irradiating a target area in a subject, e.g., a tumor or lesion, near the tumor, a lymph node, near the lymph node, or the tumor microenvironment (TME) of a tumor or lesion, with a wavelength of light to activate the phthalocyanine dye moiety of the conjugate. In some embodiments, the methods and uses provided herein include treatment of a subject having one or more tumors or lesions, e.g., one or more primary tumors or lesions (or first tumor or lesion), one or more secondary tumors or lesions (or second tumor or lesion), one or more newly occurring tumors or lesions, and / or one or more metastatic tumors or lesions. A subject may have one, two, three, or more than three tumors. Such tumors may be in one or more tissues or organs, e.g., one tissue or organ, two different tissues or organs, three different tissues or organs, or more than three different tissues or organs. In some aspects, one or more of the tumors to be treated express PD-L1 on the surface of cells contained therein. In some aspects, one or more of the tumors to be treated contain, consist primarily of, have a significant 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 significant number of, or consist entirely of cells that have a reduced response, are resistant, or become resistant (i.e., acquire resistance) to PD1 / PD-L1 checkpoint blockade.

[0261] In some aspects, the tumor or lesion treated by the provided embodiments is naïve with respect to an immune checkpoint inhibitor or has not previously been treated with an immune checkpoint inhibitor, e.g., naïve to anti-PD-1 therapy, anti-PD-L1 therapy, and / or anti-CTLA-4 therapy. In some embodiments, the tumor or lesion has not been treated with (naïve to) an anti-PD-1 treatment. In some embodiments, the tumor or lesion has not been treated with (naïve to) an anti-PD-L1 treatment. In some embodiments, the tumor or lesion has not been treated with (naïve to) an anti-CTLA-4 treatment. In some embodiments, the subject having the tumor or lesion treated by the provided embodiments is naïve with respect to an immune checkpoint inhibitor. In some embodiments, the subject being treated is naïve to an anti-PD-1 treatment. In some embodiments, the subject being treated is naïve to an anti-PD-L1 treatment. In some embodiments, the subject being treated is naïve to an anti-CTLA-4 treatment. Treatment of a subject with an immune checkpoint inhibitor, such as an anti-PD-1 antibody, can lead to CD8+ effector T cell exhaustion in the tumor, its surroundings, and / or the whole body. This can lead to the inability of CD8+ T cells to recognize and localize to the tumor, or can lead to CD8+ T cells being ineffective despite localization to the tumor or in the vicinity of the tumor, leading to checkpoint inhibitor (e.g., PD-1 / PD-L1) resistance. Thus, in some cases, ineffective or insufficient CD8+ effector T cell activity can be mitigated by avoiding immune checkpoint inhibitor treatment (e.g., anti-PD-1 treatment, anti-PD-L1 treatment, and / or anti-CTLA-4 treatment) before utilizing the provided compositions, methods, or uses.In some embodiments, the response of the tumor or lesion to the treatment herein is effected by first treating the tumor or lesion with administration of an anti-PD-L1 conjugate, followed by irradiation, and then treating the tumor or lesion with an immune checkpoint inhibitor, e.g., a therapy directed against PD-1, PD-L1, and / or CTLA-4 (e.g., an anti-PD-1 antibody, and an anti-PD-L1 antibody, and / or an anti-CTLA-4 antibody). In some embodiments, the method of treatment includes selecting a subject who has not been treated with an immune checkpoint inhibitor (e.g., an anti-PD-1, anti-PD-L1, and / or anti-CTLA-4) therapy, and treating such subject (i.e., the tumor or lesion of such subject) with an anti-PD-L1 conjugate, followed by irradiation.

[0262] In some aspects, the tumor or lesion for treatment by the provided embodiments is associated with a cancer selected from the group consisting of colon cancer, colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell cancer, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, cancer of the small intestine, spindle cell carcinoma, liver cancer, liver cancer, cancer of the peripheral nerves, brain cancer, cancer of skeletal muscle, cancer of smooth muscle, bone cancer, cancer of adipose tissue, cervical cancer, uterine cancer, cancer of the reproductive organs, lymphoma, and multiple myeloma.

[0263] In some aspects, the tumor or lesion treated by the provided embodiments includes one or more primary (e.g., first) tumors or lesions. In some aspects, the primary tumor or lesion can include the first or initial tumor or lesion in the subject. In some aspects, the subject can have one or more primary tumors or lesions. In some embodiments, the primary tumor can be a solid tumor, a lymphoma, or a leukemia. The tumor can be lung, stomach, liver, pancreas, breast, esophagus, head and neck, brain, peripheral nerve, skin, small intestine, colon, rectum, anus, ovary, uterus, bladder, prostate, adipose tissue, skeletal muscle, smooth muscle, blood vessel, bone, bone marrow, eye, tongue, lymph node, spleen, kidney, cervix, male reproductive organs, female reproductive organs, testis, or a tumor of unknown primary origin.

[0264] In some aspects, the tumor or lesion treated by the provided embodiments includes one or more second tumors or lesions, such as metastatic tumors or lesions, or newly developed tumors or lesions. In some aspects, the one or more second tumors or lesions are derived from 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 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 developed tumors or lesions. In some aspects, the one or more second tumors or lesions are derived from a different origin compared to the first tumor or lesion. In some aspects, the one or more second tumors or lesions are generated from different organs or different cells compared with the first tumor or lesion.In some embodiments, the one or more second tumors or lesions can be 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, colon, 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 reproductive organs, female reproductive organs, testis, or tumor of unknown origin.

[0265] In some embodiments, following administration of the provided anti-PD-L1 conjugate, the first tumor is irradiated and immunity to the second tumor or lesion is provided when the volume of the first tumor is reduced. In such embodiments, the volume of the first tumor is reduced by at least 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 the second tumor or lesion is provided when the first tumor achieves a partial or complete response (PR or CR) after treatment of the first tumor. In some embodiments, immunity against the second tumor or lesion is provided when the first tumor achieves a CR after treatment of the first tumor.

[0266] In some cases, the target area for irradiation may be the primary tumor or lesion or near the primary tumor or lesion, in other cases, the target area for irradiation is not the primary tumor or lesion but a different area where PD-L1 expressing cells are present, such as a lymph node, or a secondary tumor or lesion or near the secondary tumor or lesion.

[0267] In some aspects, following treatment with provided embodiments comprising anti-PD-L1 conjugate treatment and light irradiation, 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, following treatment with provided embodiments comprising anti-PD-L1 conjugate treatment and light irradiation, 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, or both tumor growth and volume are reduced. In some aspects, treatment with provided embodiments delays regrowth of a tumor or lesion, prevents recurrence of cancer, or extends the period of remission of cancer, e.g., cancer associated with the tumor or lesion, prevents or inhibits the occurrence and / or growth of one or more second tumors or lesions, e.g., second tumors or lesions of a different type compared to the primary tumor or lesion, and / or prevents or inhibits the occurrence and / or growth of metastases.

[0268] In some embodiments, the subject is administered the anti-PD-L1 conjugate to treat and / or inhibit the growth of a first tumor or first lesion; the method inhibits or delays the appearance of one or more second tumors or lesions or metastasis of the first tumor or first lesion.

[0269] In some aspects, the primary tumor or lesion contains cells expressing PD-L1 on the surface. In some aspects, the cells expressing PD-L1 are immune cells, e.g., immunosuppressive cells, e.g., M2 TAM, tDC, or MDSC. In some aspects, the PD-L1 expressing cells are tumor-associated fibroblasts or cancer-associated fibroblasts (CAFs). In some cases, the cells expressing PD-L1 are tumor cells or cancer cells. In some of any of the embodiments, the subject being treated has one or more of the PD-L1 expressing cells, e.g., one or more PD-L1 expressing cells associated with a tumor, a lesion, or a cancer.

[0270] In some embodiments, the tumor, lesion, or cancer to be treated contains tumor cells or cancer cells that do not express PD-L1. In some embodiments, the tumor or lesion comprises PD-L1 negative tumor cells. In some embodiments, more than or about 40%, 50%, 60%, 70%, 80%, 90%, or 95% of the tumor cells in the tumor or lesion are PD-L1 negative tumor cells. In some aspects, PD-L1 negative tumor cells can refer to tumor cells that do not express detectable levels of PD-L1 on their surface or tumor cells that express PD-L1 at a threshold level, e.g., below a detectable threshold level. 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 level of PD-L1 expression is determined by flow cytometry. In some aspects, the provided embodiments result in indirect killing of tumor or cancer cells, for example, by elimination of immunosuppressive cells, e.g., M2 TAMs, tDCs, or MDSCs, as well as enhancing the function and / or activity of effector cells of the immune system, e.g., CD8+ T cells, that can exert an anti-tumor or anti-cancer response to eliminate tumor or cancer cells.

[0271] In some embodiments, the tumor, lesion, or cancer being treated contains tumor or cancer cells that express PD-L1. In some aspects, administration of an anti-PD-L1 conjugate and subsequent light irradiation can indirectly kill cells that express PD-L1. In some aspects, the embodiments provided result in the direct killing of PD-L1 expressing tumor cells.

[0272] In some embodiments, the methods and uses provided herein include treating a subject with invasive tumor cells, for example when cells originating from a primary tumor have invaded surrounding tissues. The method includes administering an anti-PD-L1 conjugate to a subject with invasive tumor cells, and irradiating a target area with a wavelength appropriate for the selected phthalocyanine dye after administration of the conjugate. In some embodiments, the method includes administering an immunomodulator, such as an immune checkpoint inhibitor, before, simultaneously with, or after administration of the conjugate. In some aspects, invasive tumor cells refer to cells that arise from a primary tumor and invade the surrounding tissues of the same organ as the primary tumor or nearby organs or cavities in the body of a subject with a primary tumor.

[0273] In some cases, the methods and uses provided herein include irradiation of target area.In some aspects, the target area is one or more primary tumors, and some or all of invasive tumor cells are not irradiated, and in such a method, the growth of invasive tumor cells is inhibited, reduced or eliminated, the volume of one or more invasive tumors is reduced, or any combination thereof occurs.In some embodiments, together with the effect on one or more invasive tumor cells, the growth of primary tumor is inhibited, reduced or eliminated, and the volume of one or more primary tumors is reduced.

[0274] In some embodiments, the invasive tumor cells are contained in solid tumors. In some embodiments, the invasive tumor cells are contained in body fluids, including but not limited to ascites, pleural fluid, and cerebrospinal fluid. In some embodiments, the invasive tumor cells are contained in body cavity effusions, including but not limited to peritoneal effusion (ascites), pleural effusion, and pericardial effusion.

[0275] In some embodiments, the methods and uses provided herein include the treatment of a subject having one or more primary tumors and also having metastatic tumor cells. The method includes administering an anti-PD-L1 conjugate to a subject having a primary tumor and metastatic tumor cells, and irradiating the target area with a wavelength appropriate for the selected phthalocyanine dye after administration of the conjugate. In such a method, the growth of metastatic tumor cells is inhibited, reduced, or eliminated, the volume of one or more metastatic tumors is reduced, or any combination thereof occurs.

[0276] 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, e.g., not directly irradiated.

[0277] In some embodiments of the method and use, only the target area, such as lymph nodes or the target area that contains and / or is in the vicinity of the primary tumor or lesion, is irradiated.In some aspects, the second tumor or lesion, such as metastatic tumor or lesion, is not irradiated.

[0278] In some aspects, metastatic tumor cells include cells that originate from a primary tumor and spread to distant tissues or organs in the body of a subject with a primary tumor.Metastatic tumor cells can be located in one or more locations in the lung, stomach, liver, pancreas, breast, esophagus, head and neck, brain, peripheral nerves, skin, small intestine, colon, 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 reproductive organs, female reproductive organs, testes, blood, bone marrow, cerebrospinal fluid, or any other tissue or organ.In some embodiments, metastatic tumor cells are contained in a solid tumor.In some embodiments, metastatic tumor cells are circulating tumor cells or are not associated with a tumor mass.

[0279] In some embodiments, the method and use comprises administration of an immunomodulator, such as a checkpoint inhibitor, before, simultaneously, or after administration of the conjugate.In some embodiments, the method and use comprises administration of a second conjugate, such as a second immunoconjugate, before, simultaneously, or after administration of the conjugate provided in the present invention, followed by irradiation.In some embodiments, the method and use comprises administration of one or more additional anti-cancer treatments, such as chemotherapy, anti-angiogenic therapy, kinase inhibitor, radiation therapy, small molecule therapy, or other treatments, such as one or more of any of the treatments described in the section entitled "Combination Therapy" herein.

[0280] C. Methods and Compositions for Treating Tumors or Tumor Cells that are Hyporesponsive, Refractory, or Non-Responsive to Previous Therapeutic Treatments In some embodiments, compositions containing anti-PD-L1 conjugates, i.e., phthalocyanine dye-targeting molecule conjugates, where the targeting molecule binds to PD-L1 (e.g., anti-PD-L1 antibody-IR700 conjugates), and methods and uses comprising the anti-PD-L1 conjugates for the treatment or treatment of a tumor or cancer that has failed, been hyporesponsive, failed to achieve a desired level of response, achieved a lower than desired level of response (e.g., was inadequately responsive or therapeutically ineffective), or been non-responsive to one or more previous treatments, e.g., with an immunomodulatory agent, e.g., an immune checkpoint inhibitor, and / or an anti-cancer agent, e.g., an anti-cancer agent that indirectly targets tumor or cancer cells, are provided. In some embodiments, the tumor or cancer is one that has achieved a lower than desired level of response to, or is predicted to be resistant to, anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy. In some embodiments, the tumor or cancer has achieved a lower than desired level of response to, or is predicted to be resistant to, anti-PD-L1 therapy. In some embodiments, the tumor or cancer has achieved a lower than desired level of response to, or is predicted to be resistant to, anti-PD-1 therapy. In some embodiments, the tumor or cancer has achieved a lower than desired level of response to, or is predicted to be resistant to, anti-CTLA-4 therapy.

[0281] Cancer includes primary tumors or multiple primary tumors, and also includes metastatic tumor cells, e.g., metastatic cancers; newly occurring tumors or cancers; cancers including primary tumors or multiple primary tumors; and / or invasive tumor cells, e.g., invasive cancers. In some aspects, the provided compositions, methods, uses, and combinations can also sensitize non-inflammatory tumors, e.g., primary non-inflammatory tumors and secondary non-inflammatory tumors (e.g., metastatic tumors), to immunomodulatory agents or other anti-cancer treatments.

[0282] Such methods and uses include, for example, administering an anti-PD-L1 conjugate to a subject having a tumor or tumor cells, and then irradiating the target area (e.g., the site where PD-L1 expressing cells are present) using an appropriate light wavelength and dose for the phthalocyanine dye. In some aspects, the irradiation results in irradiation-dependent lysis and death of cells expressing the target molecule (e.g., PD-L1) on their surface, resulting in a therapeutic effect or cancer treatment. In some cases, cells expressing PD-L1, such as monocytes, macrophages, dendritic cells (DCs), M2 tumor-associated macrophages (M2 TAMs), tolerogenic dendritic cells (tDCs), or myeloid-derived suppressor cells (MDSCs), or certain tumor cells, are killed and thus rapidly depleted. As a result, necrosis of the tumor cells may occur.

[0283] In some aspects, tumors, lesions, or cancers treated by the provided embodiments include those that have failed, had a low response or were substantially non-responsive, were low-responsive, did not achieve a desired level of response, achieved a lower than desired level of response (e.g., were inadequately responsive or were not therapeutically effective), subsequently relapsed, were refractory, and / or were resistant to one or more prior treatments, e.g., with an immunomodulatory agent, e.g., an immune checkpoint inhibitor, and / or an anti-cancer agent, e.g., an anti-PD-L1 therapy, an anti-PD-1 therapy, or an anti-CTLA-4 therapy.

[0284] In some embodiments, the subject treated by the provided embodiments has been previously treated with an anti-cancer treatment and / or an immune checkpoint inhibitor. In some embodiments, the subject treated by the provided embodiments has been previously treated with an immune checkpoint inhibitor. In some embodiments, the subject treated by the provided embodiments has failed or relapsed after a previous treatment with an anti-cancer treatment and / or an immune checkpoint inhibitor. In some embodiments, the subject treated by the provided embodiments has failed or relapsed after a previous treatment with an immune checkpoint inhibitor.

[0285] In some embodiments, the inhibition of tumor growth resulting from the implementation of the method is greater than the inhibition of tumor growth as a result of a previous treatment with an anti-cancer treatment and / or immune checkpoint inhibitor (e.g., anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy). In some embodiments, the inhibition of tumor growth resulting from the implementation of the method is greater than the inhibition of tumor growth as a result of a previous treatment with an immune checkpoint inhibitor (e.g., anti-PD-L1 therapy, anti-PD-1 therapy, and / or anti-CTLA-4 therapy).

[0286] In some aspects, the past therapeutic treatment that the cancer is non-responsive to includes the use of anti-cancer drugs. The past anti-cancer drugs can be one or more of chemotherapy drugs, antibody treatment, and / or radiation therapy drugs. In some embodiments, the past treatment is a treatment with an anti-cancer drug selected from checkpoint inhibitors, immune adjuvants, chemotherapy drugs, radiation, and biologics that include anti-cancer targeting molecules that bind to tumor cells. In some embodiments, the past treatment is a treatment with an anti-cancer drug that is an antibody conjugate. In some embodiments, the past treatment is a treatment with an antibody conjugate that includes a phthalocyanine dye, a toxin, or a TLR agonist.

[0287] In some aspects, the previous therapeutic treatment to which the cancer, tumor, or tumor cell is non-responsive can be a treatment with an immune checkpoint inhibitor (also known as immune checkpoint blockade therapy). The previous immune checkpoint inhibitor can be a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, or a combination thereof. The previous immune checkpoint inhibitor can be a small molecule inhibitor, an antibody inhibitor, or other molecule that binds to and inhibits immune checkpoint proteins, such as PD-1 or PD-L1. Exemplary antibody inhibitors against PD-1 include pembrolizumab (MK-3475, Keytruda), nivolumab (OPDIVO), cemiplimab (LIBTAYO), toripalimab (JS001), HX008, SG001, GLS-010, dostarlimab (TSR-042), tislelizumab (BGB-A317), cetrelimab (JNJ-63723283), and others. ), pidilizumab (CT-011), genolimzumab (APL-501, GB226), BCD-100, cemiplimab (REGN2810), F520, sintilimab (IBI308), GLS-010, CS1003, LZM009, camrelizumab (SHR-1210), SCT-I10A, MGA012, AK105, PF-06801591, AMP-224, AB122, AMG 404, BI 754091, HLX10, JTX-4014, MEDI0680, Sym021, MGD019, MGD013, AK104, XmAb20717, RO7121661, CX-188, and spartalizumab.Exemplary antibody inhibitors against PD-L1 include atezolizumab (MPDL3280A, Tecentriq), avelumab (Bavencio), durvalumab (MEDI4736, Imfinzi), LDP, NM-01, STI-3031, KN035, LY3300054, M7824 (MSB0011359C), BMS-936559, MSB2311, BC These may include, but are not limited to, any of the following: D-135, BGB-A333, CBT-502, cosibelimab (CK-301), CS1001, FAZ053, MDX-1105, SHR-1316, TG-1501, ZKAB001, INBRX-105, MCLA-145, KN046, LY3415244, REGN3504, and HLX20.

[0288] In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that has been resistant, refractory, or non-responsive to treatment with an anti-PD-1 antibody or anti-PD-L1 antibody. In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that has been resistant, refractory, or non-responsive to treatment with an anti-PD-L1 antibody, or that is predicted to be resistant, refractory, or non-responsive 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 has been resistant, refractory, or non-responsive to treatment with an anti-PD-1 antibody, or that is predicted to be resistant, refractory, or non-responsive to treatment with an anti-PD-1 antibody.

[0289] In some aspects, the previous treatment is treatment with an anti-CTLA-4 antibody, e.g., ipilimumab (YERVOY), tremelimumab, AGEN1181, AGEN1884, ADU-1064, BCD-145, and BCD-217. In some aspects, the tumor, lesion, or cancer to be treated includes a tumor or cancer that has been resistant, refractory, or non-responsive to treatment with an anti-CTLA-4 antibody, or is predicted to be resistant, refractory, or non-responsive to treatment with an anti-CTLA-4 antibody.

[0290] In some aspects, the previous therapeutic treatment to which the cancer, tumor, or tumor cells are non-responsive may include the use of an immunomodulatory agent, such as a cytokine, e.g., aldesleukin (PROLEUKIN), interferon alpha-2a, interferon alpha-2b (INTRODUCT), or combination therapy with other agents. A), pegylated interferon alpha-2b (SYLATRON / PEG-Intron), or cytokines targeting the IFNAR1 / 2 pathway, the IL-2 / IL-2R pathway, or adjuvants such as poly ICLC (HILTONOL / Imiquimod), 4-1BB (CD137; TNFRS9), OX40 (CD134), OX40 ligand (OX40L), the Toll-like receptor 2 agonist SUP3, agonists of the Toll-like receptors TLR3 and TLR4, and adjuvants targeting the Toll-like receptor 7 (TLR7) pathway, other members of the TNFR superfamily and the TNF superfamily, other TLR2 agonists, TLR3 agonists, and TLR4 agonists.

[0291] In some aspects, the past therapeutic treatments that cancer is non-responsive to include the use of therapeutic agents that target immunosuppressive cells.The agent can be an antibody, such as an anti-CD25 antibody that targets regulatory T cells, such as basiliximab (Simulect®), daclizumab, or PC61; a small molecule inhibitor, or a combination thereof.The immunosuppressive cell includes regulatory T cells, M2 macrophages, tumor-associated fibroblasts, or cancer-associated fibroblasts (CAFs), or a combination thereof.

[0292] In some cases, the tumors, lesions, or cancers treated by the provided embodiments include "non-inflammatory tumors" or "non-inflammatory cancers," e.g., tumors with an immunosuppressive phenotype. Such non-inflammatory tumors include tumors that express intratumoral CD8 +The non-inflammatory tumor or cancer may have characteristics including, but not limited to, a substantial reduction or absence of the number and / or activity of T effector cells, and / or a substantial increase in the number and / or activity of intratumoral immunosuppressive cells. In some cases, the non-inflammatory tumor or cancer has a high tumor mutation burden (TMB), an immune score indicative of low immune responsiveness, a status (e.g., cell surface expression) of programmed cell death protein 1 (PD-1) or programmed cell death ligand 1 (PD-L1) markers that may be indicative of low immune responsiveness. In some cases, the non-inflammatory tumor or cancer does not respond to monotherapy with a PD-1 inhibitor or a PD-L1 inhibitor.

[0293] In some embodiments, non-inflammatory tumors or cancers may be treated with an anti-PD-L1 conjugate as described herein and then irradiated. In some embodiments, combined treatment with an anti-PD-L1 conjugate and subsequent irradiation with an immunomodulatory agent, such as an immune checkpoint inhibitor, results in an enhanced inhibitory effect on the growth of both the irradiated primary tumor and distant tumors.

[0294] Furthermore, for tumors that are resistant to treatment with immune-modulating therapies, e.g., treatment with immune checkpoint inhibitors, treatment with anti-PD-L1 conjugates in combination with immune checkpoint inhibitors followed by light irradiation can result in enhanced inhibitory effects on the growth of both the irradiated primary tumor and distant tumors, the primary tumor and newly arising tumors, and / or the primary tumor and different types of secondary tumors, indicating the sensitizing effect of anti-PD-L1 photoimmunotherapy to immune checkpoint inhibitors in the treatment of cancer and tumor cells.

[0295] IV. Methods of Administration and Formulations Compositions comprising the PD-L1 antibodies, antigen-binding fragments, and immunoconjugates, including pharmaceutical compositions and formulations, are also provided.

[0296] In some embodiments, the anti-PD-L1 conjugates may 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 (e.g., subcutaneous, intramuscular, intradermal, intraperitoneal, intratumoral, and intravenous), oral, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation routes. In some embodiments, the anti-PD-L1 conjugates are administered intravenously. In some embodiments, the anti-PD-L1 conjugates are administered parenterally. In some embodiments, the anti-PD-L1 conjugates are administered orally. In some embodiments, the conjugates are administered by local injection. In some embodiments, the conjugates are administered as a topical application.

[0297] A composition comprising an anti-PD-L1 conjugate may be administered locally or systemically using any method known in the art, e.g., to a subject having a tumor, e.g., cancer, or a subject from whom a tumor has previously been removed, e.g., via surgery. Specific examples are provided, but one of skill 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 over a period of hours to days to a subject in need of treatment.

[0298] In some embodiments, the anti-PD-L1 conjugate is administered intratumorally, e.g., by parenteral means, e.g., by direct injection or infusion into the tumor. In some embodiments, the anti-PD-L1 conjugate is administered to the tumor by applying the agent to the tumor, e.g., by bathing the tumor in a solution containing the anti-PD-L1 conjugate, or by pouring the agent into the tumor.

[0299] Additionally or alternatively, the anti-PD-L1 conjugate may be administered systemically, e.g., intravenously, intramuscularly, subcutaneously, intradermally, intraperitoneally, subcutaneously, or orally, to a subject with a tumor, e.g., cancer.

[0300] Compositions, e.g., pharmaceutical compositions, containing anti-PD-L1 conjugates, and uses of such compositions, e.g., therapeutic and / or pharmaceutical uses, are also provided herein. In some aspects, the composition comprises an anti-PD-L1 conjugate and a pharma- ceutically acceptable carrier. In some embodiments, the composition containing the anti-PD-L1 conjugate is for use in a treatment or therapy according to any of the provided embodiments, e.g., for administration to a subject having a disease or condition for the treatment of the disease or condition. The dosage of the anti-PD-L1 conjugate administered to a subject is not subject to absolute limitations, but will depend on the nature of the composition and its active ingredients, as well as undesired side effects, e.g., immune response to the agent, the subject being treated, and the type of condition being treated, and the mode of administration. In general, the dose will be a therapeutically effective amount, e.g., an amount sufficient to achieve a desired biological effect, e.g., an amount effective to reduce the size, e.g., volume and / or weight, of a tumor, or to attenuate further growth of a tumor, or to reduce undesirable symptoms of a tumor.

[0301] In some embodiments, compositions used for administration of the anti-PD-L1 conjugates contain an effective amount of the agent, together with 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 oral administration may contain an effective amount of the anti-PD-L1 conjugates in an aqueous solution or suspension that may optionally include buffers, surfactants, thixotropic agents, and flavoring agents.

[0302] In some embodiments, the anti-PD-L1 conjugate, or the conjugate combined with an additional therapeutic agent, may be formulated in a pharma- ceutically acceptable buffer, such as one that contains a pharma- ceutically acceptable carrier or vehicle. In general, the pharma- ceutically acceptable carrier or vehicle, such as one present in a pharma- ceutically acceptable buffer, may be any known in the art. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 19th Edition (1995), describes compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compounds. Pharmaceutically acceptable compositions are generally prepared with consideration for approval by regulatory or other authorities and in accordance with generally recognized pharmacopoeias for use in animals and humans.

[0303] A pharmaceutical composition may include a carrier, such as a diluent, adjuvant, excipient, or vehicle, for administering 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, together with a suitable amount of carrier to provide the form for proper administration to the patient. Such pharmaceutical carriers can be sterile liquids, such as water, and oils, such as those 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. Saline solutions and aqueous dextrose and glycerol solutions can also be utilized as liquid carriers, particularly for injectable solutions. The compositions may contain, together with the active ingredient, diluents such as lactose, sucrose, calcium hydrogen phosphate, or carboxymethylcellulose; lubricants such as magnesium stearate, calcium stearate, and talc; and binders such as starch, natural gums such as gum arabic, 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, dried skim milk, glycerol, propylene, glycol, water, and ethanol. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, such as acetic acid, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and other such agents.

[0304] In some embodiments, pharmaceutical preparations can be liquid, for example, solution, syrup or suspension.Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives, for example, suspending agents (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (for example, lecithin or gum arabic); non-aqueous medium (for example, almond oil, oily esters or fractionated vegetable oils); and preservatives (for example, methyl-p-hydroxybenzoate or propyl-p-hydroxybenzoate or sorbic acid).In some cases, pharmaceutical preparations can be presented in lyophilized form for reconstitution with water or other suitable medium before use.

[0305] In some embodiments, the nature of the pharma- ceutically acceptable buffer or carrier depends on the particular mode of administration to be used. For example, in some embodiments, parenteral formulations may contain injectable fluids that contain pharma- ceutically and physiologically acceptable fluids, such as water, physiological saline, balanced salt solutions, aqueous dextrose, or glycerol as a vehicle. In some embodiments, non-toxic solid carriers for solid compositions, such as powder, pill, tablet, or capsule forms, may include, for example, pharmaceutical grades of 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, citric acid, and other organic acids; antioxidants, such as ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl alcohol or benzyl alcohol; alkyl parabens, such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, Examples of suitable carriers include, but are not limited to, proteins such as serum albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates such as glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; counterions that form salts such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as polyethylene glycol (PEG). In addition to biologically neutral carriers, the pharmaceutical compositions to be administered may contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents, such as sodium acetate or sorbitan monolaurate, in some embodiments.

[0306] A buffering agent is included in the composition in some aspects. Suitable buffering agents 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 buffering agents is used. The buffering agent or mixture is typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0307] Formulations of the antibodies described herein may include lyophilized formulations and aqueous solutions.

[0308] The formulation or composition may contain multiple active ingredients useful for a particular indication, disease, or condition treated by the antibody, antigen-binding fragment, or conjugate, preferably those having complementary activities to the antibody, antigen-binding fragment, or conjugate, and each activity does not adversely affect each other. Such active ingredients are suitably present in combination in an amount effective for the intended purpose. Thus, in some embodiments, the pharmaceutical composition further comprises other pharmacoactive agents or drugs, such as chemotherapeutic agents, such as asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0309] Compound can be formulated into suitable pharmaceutical preparations for oral administration, such as solution, suspension, tablet, dispersible tablet, pill, capsule, powder, sustained release preparation or elixir, or can be formulated into transdermal patch preparation and dry powder inhaler.Typically, compound is formulated into pharmaceutical composition by using techniques and methods well known in the art (for example, see Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition, 1985, 126).Generally, the mode of formulation varies according to the route of administration.

[0310] In some aspects, pharmaceutical compositions can utilize time-released, sustained release, and sustained release delivery systems, so that the delivery of the composition occurs before the site to be treated is sufficiently long to cause the site to become sensitive.Many types of release delivery systems are available and known.Such systems can avoid repeated administration of the composition, thereby increasing the convenience for the subject and the physician.

[0311] In some embodiments, the pharmaceutical composition contains an antibody, antigen-binding fragment, or conjugate in an amount effective for treating or preventing a disease or condition, e.g., a therapeutically effective amount or a prophylactically effective amount. The therapeutic or prophylactic effectiveness is monitored in some embodiments by periodic assessment of the subject being treated. In the case of repeated administration over several days or more, depending on the condition, the treatment is repeated until the desired suppression of the symptoms of the disease occurs. However, other dosing schedules may be useful and can be determined. The desired dosage can be delivered by a single bolus administration of the composition, by multiple boluses of the composition, or by continuous infusion administration of the composition.

[0312] The composition can be formulated for administration by any route known to those skilled in the art, such as intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, intranasal, oral, vaginal, rectal, local, topical, otic, inhalation, buccal (e.g., sublingual), and transdermal administration, or any other route. Other modes of administration are also contemplated. Administration can be topical, localized, or systemic, depending on the location of treatment. Local administration to the area requiring treatment can be achieved, for example, but not limited to, by topical application, injection, catheter, suppository, or implant, combined with local injection during surgery, e.g., wound dressing after surgery.

[0313] Parenteral administration, generally characterized by injection, either subcutaneously, intramuscularly, intratumorally, intravenously, or intradermally, is contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solids suitable for dissolving or suspending in liquid prior to injection, or emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the administered pharmaceutical composition may also contain an activator in the form of a solvent, for example, a pH buffer, a metal ion salt, or other such buffer. The pharmaceutical composition may also contain minor amounts of other non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffers, stabilizers, solubility enhancers, and other such agents, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins. Implantation of a sustained-release or sustained-release system such that a constant dosage level is maintained (see, for example, U.S. Pat. No. 3,710,795) is also contemplated herein. The percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject.

[0314] Injectables are designed for local and systemic administration. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products ready for combination with a solvent immediately before use, such as lyophilized powders, such as tablets for subcutaneous injection, sterile suspensions ready for injection, sterile dry insoluble products ready for combination with a vehicle immediately before use, and sterile emulsions. The solutions can be either aqueous or non-aqueous. When administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), as well as solutions containing thickening agents and solubilizers, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0315] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous media, non-aqueous media, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharma- ceutically acceptable substances. Examples of aqueous media include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, and lactated Ringer's dextrose injection. Non-aqueous parenteral media include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil, and peanut oil. Parenteral preparations packaged in multi-dose containers may contain antimicrobial agents in bacteriostatic or fungistatic concentrations, such as phenol or cresol, mercurials, benzyl alcohol, chlorobutanol, p-hydroxybenzoic acid methyl and propyl esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate.

[0316] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), as well as solutions containing thickening agents and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0317] The composition may be formulated for single administration or for multiple administration.The drug may be formulated for direct administration.The composition may be provided as a liquid or lyophilized formulation.When the composition is provided in lyophilized form, it can be reconstituted with a suitable buffer, for example, sterile saline solution, immediately before use.

[0318] The compositions may be administered continuously, intermittently, or in the same composition with other biologically active agents. Administration can also include controlled release systems, such as controlled release formulations, and controlled release by devices, such as by pumps.

[0319] The most suitable route in a particular case depends on various 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 methods can also be used, but compared to intravenous methods, increased absorption times may be required to ensure equivalent bioavailability.

[0320] Pharmaceutical compositions can be formulated in suitable dosage forms for each administration route. Pharmaceutically and therapeutically active compounds and their derivatives are typically formulated and administered in unit or multiple dosage forms. Each unit dosage contains a predetermined amount of therapeutically active compound sufficient to produce the desired therapeutic effect, together with the required 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, and oil-water emulsions, containing the compound or its pharma-ceutically acceptable derivatives in an appropriate amount. Unit dosage forms may be contained in ampoules and syringes, or may be packaged individually in tablets or capsules. Unit dosage forms may be administered in divided doses or in multiples. Multiple dosage forms are a plurality of identical unit dosage forms packaged in a single container to be administered in separate unit dosage forms. Examples of multiple dose forms include vials, bottles of tablets or capsules, or bottles of pints or gallons. Thus, multiple dose forms are a plurality of unit doses that are not separated in packaging. Generally, dosage forms or compositions containing active ingredients in the range of 0.005% to 100%, the remainder being composed of non-toxic carriers, can be prepared. Pharmaceutical compositions can be formulated into dosage forms appropriate for each administration route.

[0321] The concentration of the pharmaceutically active compound is adjusted to provide an effective amount for injection to produce the desired pharmacological effect. The exact dose depends on the age, weight, and condition of the patient or animal, as known in the art. Unit dose parenteral preparations are packaged in ampoules, vials, or syringes with needles. The volume of liquid solution or reconstituted powder preparation containing the pharmaceutically active compound varies depending on the disease to be treated and the specific product selected for the package. As known and practiced in the art, all preparations for parenteral administration must be sterile. In some embodiments, the compositions can be provided as lyophilized powders that can be reconstituted for administration as solutions, emulsions, and other mixtures. They can also be reconstituted and formulated as solids or gels. Lyophilized powders can be prepared from any of the above solutions.

[0322] A sterile, lyophilized powder can be prepared by dissolving the phthalocyanine dye-targeting molecule conjugate in a buffer solution that may contain excipients that improve the stability of other pharmacological components of the powder, or of a reconstituted solution prepared from the powder.

[0323] In some embodiments, subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation. Briefly, lyophilized powders are prepared by dissolving an excipient, such as dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agent, in a suitable buffer, such as citric acid, sodium or potassium phosphate, or other such buffer known to those of skill in the art. The selected enzyme is then added to the resulting mixture and stirred until it dissolves. The resulting mixture is sterile filtered or otherwise treated to remove particulates and ensure sterility, and dispensed into vials for lyophilization. Each vial may contain a single dose (1 mg to 1 g, typically 1 to 100 mg, e.g., 1 to 5 mg) or multiple doses of the compound. The lyophilized powder may be stored under appropriate conditions, e.g., at about 4° C. to room temperature. Reconstitution of this lyophilized powder with a buffer solution provides a formulation for use in parenteral administration. The exact amount depends on the indication being treated and the compound selected. Such amounts can be determined empirically.

[0324] In some embodiments, the pH of the composition is 6-10, or about 6-10, e.g., 6-8, or about 6-8, 6.9-7.3, or about 6.9-7.3, e.g., about pH 7.1. In some embodiments, the pH of the pharma- ceutically 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.

[0325] The compositions may be formulated for single or multiple administration. The agents may be formulated for direct administration.

[0326] In some embodiments, the compositions provided herein comprise a concentration of from 0.01 mg or about 0.01 mg to 3000 mg or about 3000 mg, from 0.01 mg or about 0.01 mg to 1000 mg or about 1000 mg, from 0.01 mg or about 0.01 mg to 500 mg or about 500 mg, from 0.01 mg or about 0.01 mg to 100 mg or about 100 mg, from 0.01 mg or about 0.01 mg to 50 mg or about 50 mg, from 0.01 mg or about 0.01 mg to 10 mg or about 1 ... or from about 0.01 mg, 1 mg or about 1 mg, 0.01 mg or about 0.01 mg to 0.1 mg or about 0.1 mg, 0.1 mg or about 0.1 mg to 2000 mg or about 2000 mg, 0.1 mg or about 0.1 mg to 1000 mg or about 1000 mg, 0.1 mg or about 0.1 mg to 500 mg or about 500 ...

Claims

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

34. The antibody or antigen-binding fragment thereof, comprising:

2. (a) the VH region comprises CDR-H1 having the amino acid sequence set forth in SEQ ID NO:87, CDR-H2 having the amino acid sequence set forth in SEQ ID NO:88, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO:84; and the VL region comprises CDR-L1 having the amino acid sequence set forth in SEQ ID NO:246, CDR-L2 having the amino acid sequence set forth in SEQ ID NO:247, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO:248; (b) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 40, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 41, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 37; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 210, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 211, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 212; (c) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 37; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 218, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 211, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 212; (d) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 63, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 64, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 60; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 221, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 222, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 223; (e) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 63, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 64, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 60; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 221, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 222, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 223; (f) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 75, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 76, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 60; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 229, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 222, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 223; (g) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 40, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 41, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 37; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 233, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 234, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 235; (h) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 37; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 241, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 234, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 242; (i) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 98, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 99, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 84; the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 246, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 254, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 255; (j) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 108, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 105; the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 258, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 260; (k) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 121, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 122, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 118; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 265, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 266, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 267; (l) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 134, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 131; the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 246, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 273, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 274; (m) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 147, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 148, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 144; the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 278, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 279, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 280; (n) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 160, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 161, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 157; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 286, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 287, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 288; (o) the VH region comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 173, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 174, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 170; and the VL region comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 294, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 234, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 295; (p) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 185, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 186, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 182; the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 299, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 300, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 301; (q) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 173, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 174, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 193; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 306, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 234, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 307; (r) the VH region comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 160, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 161, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 157; and the VL region comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 311, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 312, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 313; or (s) the VH region comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 202, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 203, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 199; and the VL region comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 319, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 320, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 321; The antibody or antigen-binding fragment of claim 1.

3. (a) the VH region comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:6, or comprises the amino acid sequence set forth 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 set forth in SEQ ID NO:23, or comprises the amino acid sequence set forth in SEQ ID NO:23; (b) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:1, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:17; (c) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:2, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:18; (d) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:3, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:19; (e) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:4, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:19; (f) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:5, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:20; (g) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:1, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:21; (h) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:2, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:22; (i) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:7, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:24; (j) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:8, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:25; (k) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:9, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:26; (l) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 10, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 27; (m) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:11, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:28; (n) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 12, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 29; (o) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 13, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 30; (p) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 14, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 31; (q) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 15, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 32; (r) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:12, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO:33; or (s) the VH region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 16, and the VL region comprises an amino acid sequence having at least 95% sequence identity to, or comprises the amino acid sequence set forth in SEQ ID NO: 34; The antibody or antigen-binding fragment of claim 1.

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

5. 2. The antibody or antigen-binding fragment of claim 1, comprising a human immunoglobulin Fc region and / or a human antibody framework region.

6. 2. The antibody or antigen-binding fragment of claim 1, which is a single-chain antibody fragment, a single-domain antibody, or a complete or intact antibody.

7. 2. The antibody or antigen-binding fragment of claim 1, which is a bispecific antibody that further specifically binds to a second antigen, optionally wherein the second antigen is an antigen expressed in a tumor cell or an immune cell.

8. an Fc region exhibiting one or more Fc-mediated effector functions; an Fc region that lacks Fc-mediated effector functions; an Fc region that exhibits substantially reduced Fc-mediated effector function, or Fc region that does not exhibit substantial Fc-mediated effector functions 2. The antibody or antigen-binding fragment of claim 1, comprising:

9. A conjugate comprising the antibody or antigen-binding fragment of claim 1 and a heterologous molecule or moiety.

10. 10. The conjugate of claim 9, wherein the heterologous molecule or moiety is a protein, peptide, nucleic acid, dye, or small molecule, and optionally a toxin, radioisotope, chemotherapeutic agent, lytic peptide, cytokine, or photoactivatable dye.

11. 11. The conjugate of claim 10, wherein the photoactivatable dye is a Si-phthalocyanine dye.

12. 12. The conjugate of claim 11, wherein the Si-phthalocyanine dye is IR700.

13. The phthalocyanine dye is represented by formula (I):

12. The conjugate of claim 11, having the structure:

14. 12. The conjugate of claim 11, which is activated to effect cell killing by irradiation at wavelengths at or about 600 nm to 850 nm or about 850 nm.

15. 15. The conjugate of claim 14, wherein the activated conjugate is effective in inhibiting tumor growth or killing with a higher level, activity, or potency than the unconjugated antibody.

16. A VH region comprising: i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 84; and ii) a VL region comprising CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 246, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 247, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 248; an antibody or antigen-binding fragment that specifically binds to a programmed cell death ligand (PD-L1) protein, comprising: b) a heterogeneous molecule or moiety that is a Si-phthalocyanine dye , a conjugate comprising:

17. The Si-phthalocyanine dye is IR700 or a compound represented by formula (I):

17. The conjugate of claim 16, having the structure:

18. A polynucleotide encoding the antibody or antigen-binding fragment of any one of claims 1 to 8.

19. A vector comprising the polynucleotide of claim 18.

20. 20. A modified cell comprising the vector of claim 19.

21. A modified cell expressing the antibody or antigen-binding fragment of any one of claims 1 to 8.

22. 18. A composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 8, or the conjugate of any one of claims 9 to 17, and optionally further comprising a pharmaceutically acceptable excipient.

23. 23. The composition of claim 22 for treating a tumor or cancer in a subject.

24. A medicament for use in a method for treating a disease or disorder in a subject, comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 8, the method comprising administering the antibody or antigen-binding fragment thereof to the subject; Optionally, the disease or disorder is a tumor or cancer. The medicine.

25. A medicament for use in a method of treating a disease or disorder in a subject, comprising the conjugate of any one of claims 9 to 17, the method comprising administering the conjugate to the subject; Optionally, the disease or disorder is a tumor or cancer. The medicine.

26. A medicament for use in a method of treating a disease or disorder in a subject, comprising the composition of claim 22, the method comprising administering the composition to the subject; Optionally, the disease or disorder is a tumor or cancer. The medicine.

27. A medicament for use in a method for treating a tumor or lesion in a subject, comprising the conjugate of any one of claims 9 to 17, The method comprises: (a) administering the conjugate to the subject; and (b) applying 25 J / cm 2 to a target area in the subject at a wavelength of at or about 600 nm to at or about 850 nm. 2 Or about 25J / cm 2 From 400J / cm 2 or approximately 400 J / cm 2 or at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, thereby activating the conjugate. whereby the growth, volume, or size of said tumor or lesion is reduced or inhibited; The medicine.

28. A medicament for use in a method of treating a tumor or lesion in a subject, comprising the composition of claim 22, The method comprises: (a) administering the composition to the subject; and (b) irradiating a target area in the subject with light at a wavelength of at or about 600 nm to at or about 850 nm at a dose of at or about 25 J / cm2 to at or about 400 J / cm2, or at or about 2 J / cm fiber length to at or about 500 J / cm fiber length, thereby activating the conjugate in the composition. whereby the growth, volume, or size of said tumor or lesion is reduced or inhibited; The medicine.

29. The pharmaceutical of claim 25, wherein in the method, the subject is administered the conjugate to treat, inhibit the growth of, and / or reduce the size of a first tumor or lesion; and the pharmaceutical inhibits, delays, or prevents the appearance, growth, or establishment of one or more second tumors or lesions located distal to the first tumor or lesion.

30. A medicament for use in a method of immunizing a subject having a first tumor or lesion, comprising the conjugate of any one of claims 9 to 17, The method comprises: (a) administering the conjugate to the subject; and (b) 25 J / cm at wavelengths at or about 600 nm to 850 nm or about 850 nm 2 Or about 25J / cm 2 From 400J / cm 2 or approximately 400 J / cm 2 irradiating a target area in the first tumor or lesion at a dose of at or about 2 J / cm fiber length to at or about 500 J / cm fiber length. wherein the growth of said first tumor or lesion is inhibited and / or reduced in size; and the appearance, growth, or establishment of one or more second tumors or lesions located distal to said treated first tumor or lesion is inhibited, delayed, or prevented. The medicine.

31. A pharmaceutical for use in a method of immunizing a subject having a first tumor or lesion, comprising the composition of claim 22, The method comprises: (a) administering the composition to the subject; and (b) irradiating a target area in the first tumor or lesion with a wavelength of at or about 600 nm to at or about 850 nm at a dose of at or about 25 J / cm 2 to at or about 400 J / cm 2 , or at or about 2 J / cm fiber length to at or about 500 J / cm fiber length. wherein the growth of said first tumor or lesion is inhibited and / or reduced in size; and the appearance, growth, or establishment of one or more second tumors or lesions located distal to said treated first tumor or lesion is inhibited, delayed, or prevented. The medicine.

32. 25. The pharmaceutical agent of claim 24, wherein the tumor, lesion, or cancer is associated with a cancer selected from the group consisting of colon cancer, colorectal cancer, pancreatic cancer, breast cancer, skin cancer, lung cancer, non-small cell lung cancer, renal cell cancer, thyroid cancer, prostate cancer, head and neck cancer, gastrointestinal cancer, stomach cancer, cancer of the small intestine, spindle cell neoplasm, liver cancer, liver cancer, cancer of the peripheral nerves, brain cancer, cancer of skeletal muscle, cancer of smooth muscle, bone cancer, cancer of adipose tissue, cervical cancer, uterine cancer, cancer of the reproductive organs, lymphoma, and multiple myeloma.

33. The pharmaceutical composition of claim 25, wherein the tumor, lesion, or cancer is associated with a cancer selected from the group consisting of colon 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, reproductive organ cancer, lymphoma, and multiple myeloma.

34. The pharmaceutical composition of claim 26, wherein the tumor, lesion, or cancer is associated with a cancer selected from the group consisting of colon 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, peripheral nerve cancer, brain cancer, skeletal muscle cancer, smooth muscle cancer, bone cancer, adipose tissue cancer, cervical cancer, uterine cancer, reproductive organ cancer, lymphoma, and multiple myeloma.

35. The pharmaceutical described in claim 24, wherein the method further comprises a step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

36. The pharmaceutical described in claim 25, wherein the method further comprises the step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

37. The pharmaceutical described in claim 26, wherein the method further comprises a step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

38. The pharmaceutical described in claim 27, wherein the method further comprises the step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

39. The pharmaceutical described in claim 28, wherein the method further comprises the step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

40. The pharmaceutical described in claim 30, wherein the method further comprises a step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

41. The pharmaceutical described in claim 31, wherein the method further comprises a step of administering an additional therapeutic agent or anti-cancer treatment to the subject.

42. A VH region comprising: i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 84; and ii) a VL region comprising CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 246, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 247, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 248; an antibody or antigen-binding fragment that specifically binds to a programmed cell death ligand (PD-L1) protein, comprising: b) IR700 Si-phthalocyanine dye 1. A medicament for use in a method of killing PD-L1 expressing cells in a subject, comprising a conjugate comprising: The method comprises: 1) administering the conjugate to the subject; and 2) activating the conjugate by irradiating a target area in the subject containing PD-L1-expressing cells to be killed with light of 690 nm ± 40 nm. thereby killing PD-L1 expressing cells. The medicine.

43. A VH region comprising: i) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 84; and ii) a VL region comprising CDR-L1 having the amino acid sequence set forth in SEQ ID NO: 246, CDR-L2 having the amino acid sequence set forth in SEQ ID NO: 247, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO: 248; an antibody or antigen-binding fragment that specifically binds to a programmed cell death ligand (PD-L1) protein, comprising: b) Formula (I): or a salt, stereoisomer, or tautomer thereof.

1. A medicament for use in a method of killing PD-L1 expressing cells in a subject, comprising a conjugate comprising: The method comprises: 1) administering the conjugate to the subject; and 2) activating the conjugate by irradiating a target area in the subject containing PD-L1-expressing cells to be killed with light of 670 nm ± 50 nm. thereby killing PD-L1 expressing cells. The medicine.

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