Binding members to PD-L1
The present invention relates to anti-PD-L1 binding members and in particular to monovalent, high potency PD-L1-binding antibody fragments being highly stable and soluble. Such binding members may be used in the treatment of cancer and inflammatory diseases as well as in diagnostics. Also provided are related nucleic acids, vectors, cells, and compositions.
1. A binding member having a binding specificity to PD-L1, comprising
(i) the variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID NOs: 6, 7 and 8; and
(ii) the variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID NOs: 3, 4 and 5,
wherein said binding member is chemically modified.
2. The binding member of claim 1 , being humanized.
3. The binding member of claim 1 , further comprising a linker sequence set forth in SEQ ID NO: 10.
4. The binding member of claim 1 , wherein said binding member is multivalent.
5. The binding member of claim 1 , wherein said binding member comprises an Fc domain.
6. The binding member of claim 5 , wherein said binding member comprises a constant region selected from the group consisting of a human IgG1, IgG2, IgG3, and IgG4 isotype.
7. The binding member of claim 5 , wherein said binding member comprises a constant region selected from the group consisting of a murine IgG1, IgG2A, IgG2B, and IgG3 isotype.
8. The binding member of claim 5 , wherein said Fc domain is modified and does not induce a cytotoxic immune response.
9. The binding member of claim 1 , wherein said chemical modification is PEGylation, HESylation, or conjugation to a second moiety.
10. A method of producing a binding member of having a binding specificity to PD-L1, the method comprising:
(i) cultivating a host cell comprising a nucleic acid molecule comprising a sequence encoding a binding member having a binding specificity to PD-L1 and comprising the variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID NOs: 6, 7 and 8, and the variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID NOs: 3, 4 and 5, thereby allowing the binding member to be expressed; and
(ii) recovering the binding member.
11. A method of producing a binding member having a binding specificity to PD-L1, the method comprising:
(a) contacting a cell-free expression system with a nucleic acid product template, the nucleic acid product template encoding a binding member having a binding specificity to PD-L1 and comprising the variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID NOs: 6, 7 and 8, and the variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID NOs: 3, 4 and 5 according to claim 1 ;
(b) allowing transcription and translation of the nucleic acid product template to occur, thereby allowing a reaction mixture to be formed;
(c) recovering the binding member from the reaction mixture.
12. The method of producing a binding member of claim 10 , further comprising purifying said binding member.
13. A method of detecting the presence of PD-L1 in a biological sample, the method comprising:
(a) contacting the biological sample with a binding member having a binding specificity to PD-L1 and comprising the variable heavy chain CDR-H1, CDR-H2 and CDR-H3 sequences as set forth in SEQ ID NOs: 6, 7 and 8, and the variable light chain CDR-L1, CDR-L2 and CDR-L3 sequences as set forth in SEQ ID NOs: 3, 4 and 5 under conditions permissive for specific binding of the binding member to PD-L 1 , and
(b) detecting whether a complex between the binding member and PD-L1 is formed.
14. The method of claim 13 , wherein said method is an in vitro method or an in vivo method.
15. The method of claim 13 , wherein said biological sample is of human origin.
16. The method of claim 13 , wherein said biological sample is at least one of a blood sample, a urine sample, a cerebrospinal fluid sample, a biopsy sample, or a lymph sample.
17. The method of claim 13 , wherein said method is a method for selecting subjects eligible for therapy with a binding member having a binding specificity to PD-L1.
18. The binding member of claim 4 , wherein said multivalent binding member is multispecific, bispecific, a diabody, a single chain diabody, a DART, a BiTE, or a tandem scFv.