Multispecific antibody constructs

1. A multi-specific antibody molecule comprising

a) a polypeptide chain of formula (I):

V H —CH 1 —X—V 1 ; and

b) a polypeptide chain of formula (II):

V L —C L —Y—V 2 ;

wherein:

V H represents a heavy chain variable domain;

CH 1 represents a domain of a heavy chain constant region;

X represents a bond or linker;

Y represents a bond or linker;

V 1 represents a dsscFv;

V L represents a light chain variable domain;

C L represents a domain from a light chain constant region;

V 2 represents a dsscFv.

2. The multi-specific antibody molecule according to claim 1 , wherein X is a linker.

3. The multi-specific antibody molecule according to claim 1 , wherein Y is a linker.

4. The multi-specific antibody molecule according to claim 1 , wherein the light chain variable domain of V 1 is attached to X.

5. The multi-specific antibody molecule according to claim 1 , wherein the light chain variable domain of V 2 is attached to Y.

6. The multi-specific antibody molecule according to claim 1 , wherein the light chain and heavy chain variable domains of V 1 and/or the light chain and heavy chain variable domains of V 2 are linked by a disulfide bond between two engineered cysteine residues.

7. The multi-specific antibody molecule according to claim 1 , wherein the heavy chain and light chain variable domains of V 1 and/or V 2 are linked by a disulfide bond between two engineered cysteine residues, wherein the position of the pair of engineered cysteine residues is selected from the group comprising or consisting of: V H 37 and V L 95, V H 44 and V L 100, V H 44 and V L 105, V H 45 and V L 87, V H 100 and V L 50, V H 100b and V L 49, V H 98 and V L 46, V H 101 and V L 46, V H 105 and V L 43 and V H 106 and V L 57, wherein the V H and V L values are independently selected within a given V 1 or V 2 , wherein the numbering is according to Kabat.

8. The multi-specific antibody molecule according to claim 7 , wherein the position of the pair of engineered cysteine residues is V H 44 and V L 100, wherein the numbering is according to Kabat.

9. The multi-specific antibody molecule according to claim 1 , wherein X is a peptide linker.

10. The multi-specific antibody molecule according to claim 1 , wherein Y is a peptide linker.

11. The multi-specific antibody molecule according to claim 1 which is tri-specific.

12. The tri-specific antibody molecule according to claim 11 , wherein VH/VL, V1 and V2 are each binding domains that bind to different antigens.

13. A pharmaceutical composition comprising a multi-specific antibody molecule according to claim 1 and at least one excipient.

14. The multi-specific antibody molecule according to claim 9 , wherein X is a peptide linker selected from SEQ ID NO: 1, 2, 69, or 70.

15. The multi-specific antibody molecule according to claim 10 , wherein Y is a peptide linker selected from SEQ ID NO: 1, 2, 69, or 70.

16. A formulation comprising the multi-specific antibody molecule of claim 1 , wherein at least 98% of the multi-specific antibody molecule in the formulation is present in monomeric form after storage for 28 days at 4° C.

17. The multi-specific antibody of claim 1 , wherein V 1 and V 2 are specific for two different antigens.

18. A multi-specific antibody molecule consisting of:

c) a polypeptide chain of formula (I):

V H —CH 1 —X—V 1 ; and

d) a polypeptide chain of formula (II):

V L —C L —Y—V 2 ;

wherein:

V H represents a heavy chain variable domain;

CH 1 represents a domain of a heavy chain constant region;

X represents a bond or linker;

Y represents a bond or linker;

V 1 represents a dsscFv;

V L represents a light chain variable domain that binds the first antigen;

C L represents a domain from a light chain constant region;

V 2 represents a dsscFv; and

wherein V1 and V2 are specific for two different antigens.

19. A formulation comprising a multi-specific antibody molecule comprising:

e) a polypeptide chain of formula (I):

V H —CH 1 —X—V 1 ; and

f) a polypeptide chain of formula (II):

V L —C L —Y—V 2 ;

wherein:

V H represents a heavy chain variable domain;

CH 1 represents a domain of a heavy chain constant region;

X represents a bond or linker;

Y represents a bond or linker;

V 1 represents a dsscFv;

V L represents a light chain variable domain;

C L represents a domain from a light chain constant region;

V 2 represents a dsscFv; and

wherein at least 98% of the multi-specific antibody molecule in the formulation is present in monomeric form after storage for 28 days at 4° C.

20. The multi-specific antibody molecule of claim 1 , wherein only one of V H /V L , V 1 or V 2 has specificity for a serum carrier protein.

21. The multi-specific antibody molecule of claim 20 , wherein the serum carrier protein is human serum albumin and wherein said V H /V L or V 1 or V 2 having specificity for a serum carrier protein forms an albumin binding site.

22. The multi-specific antibody molecule of claim 21 , wherein the albumin binding site comprises SEQ ID NO: 71 for CDRH1, SEQ ID NO: 72 for CDRH2, SEQ ID NO: 73 for CDRH3, SEQ ID NO: 74 for CDRL1, SEQ ID NO: 75 for CDRL2 and SEQ ID NO: 76 for CDRL3; or a heavy chain variable domain selected from SEQ ID NO: 77 and SEQ ID NO: 78 and a light chain variable domain selected from SEQ ID NO: 79 and SEQ ID NO: 80.

23. The multi-specific antibody molecule of claim 22 , wherein V 1 comprises the albumin binding site.

24. The multi-specific antibody molecule of claim 22 , wherein V 2 comprises the albumin binding site.

25. The multi-specific antibody molecule of claim 1 , wherein the heavy chain variable domain of V 1 is attached to X.

26. The multi-specific antibody molecule of claim 1 , wherein the heavy chain variable domain of V 2 is attached to Y.