Anti-CD47 x anti-mesothelin antibodies and methods of use thereof

1. An isolated bispecific antibody comprising a first arm that specifically binds CD47 and a second arm that binds mesothelin (MSLN), wherein the bispecific antibody comprises two heavy chains each comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO: 225, a CDRH2 comprising the amino acid sequence of SEQ ID NO: 226, a CDRH3 comprising the amino acid sequence of SEQ ID NO: 227,

one kappa light chain comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 240, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 242, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 254, and

one lambda light chain comprising a combination of a CDRL1 amino acid sequence, a CDRL2 amino acid sequence, and a CDRL3 amino acid sequence selected from the group consisting of:

(a) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 286, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 292, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 298;

(b) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 282, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 288, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 294;

(c) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 283, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 289, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 295;

(d) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 284, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 290, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 296;

(e) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 285, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 291, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 297;

(f) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 287, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 293, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 299; and

(g) a CDRL1 comprising the amino acid sequence of SEQ ID NO: 282, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 288, and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 300.

2. The isolated bispecific antibody of claim 1 , wherein the bispecific antibody inhibits interaction between human CD47 and human signal-regulatory protein alpha (SIRPα).

3. The isolated bispecific antibody of claim 1 , wherein the first arm comprises a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 114 and a variable light chain comprising the amino acid sequence of SEQ ID NO: 168.

4. The isolated bispecific antibody of claim 1 , wherein the first arm comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 2 and a light chain comprising the amino acid sequence of SEQ ID NO: 56.

5. The isolated bispecific antibody of claim 1 , wherein isolated bispecific antibody comprises a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 114, a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 212, 214, 216, 218, 220, 222, and 224.

6. The isolated bispecific antibody of claim 1 , wherein isolated bispecific antibody comprises a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 114, and a combination of a kappa variable light chain and a lambda light chain selected from the group consisting of

(a) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 212;

(b) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 214;

(c) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 216;

(d) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 218;

(e) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 220;

(f) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 222; and

(g) a kappa variable light chain comprising the amino acid sequence of SEQ ID NO: 168, and a lambda variable light chain comprising the amino acid sequence of SEQ ID NO: 224.

7. The isolated bispecific antibody of claim 1 , wherein isolated bispecific antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 2, a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 98, 100, 102, 104, 106, 108, and 110.

8. The isolated bispecific antibody of claim 1 , wherein isolated bispecific antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a combination of a kappa light chain and a lambda light chain selected from the group consisting of

(a) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 98;

(b) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 100;

(c) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 102;

(d) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 104;

(e) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 106;

(f) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 108; and

(g) a kappa light chain comprising the amino acid sequence of SEQ ID NO: 56, and a lambda light chain comprising the amino acid sequence of SEQ ID NO: 110.

9. The isolated bispecific antibody of claim 1 , wherein the bispecific antibody comprises two copies of a single heavy chain polypeptide and a first light chain and a second light chain, wherein the first and second light chains are different.

10. The isolated bispecific antibody of claim 9 , wherein at least a portion of the first light chain is of the Kappa type and at least a portion of the second light chain is of the Lambda type.

11. The isolated bispecific antibody of claim 10 , wherein the first light chain comprises at least a Kappa constant region.

12. The isolated bispecific antibody of claim 11 , wherein the first light chain further comprises a Kappa variable region.

13. The isolated bispecific antibody of claim 11 , wherein the first light chain further comprises a Lambda variable region.

14. The isolated bispecific antibody of claim 10 , wherein the second light chain comprises at least a Lambda constant region.

15. The isolated bispecific antibody of claim 14 , wherein the second light chain further comprises a Lambda variable region.

16. The isolated bispecific antibody of claim 14 , wherein the second light chain further comprises a Kappa variable region.

17. The isolated bispecific antibody of claim 10 , wherein the first light chain comprises a Kappa constant region and a Kappa variable region, and wherein the second light chain comprises a Lambda constant region and a Lambda variable region.

18. The isolated bispecific antibody of claim 1 , wherein the constant and variable framework region sequences are human.