Assembly of bispecific antibodies

Patent metadata

Field	Value
Publication number	US-10626189-B2
Application number	US-201715789775-A
Country	US
Kind code	B2
Filing date	Oct 20, 2017
Priority date	Oct 11, 2011
Publication date	Apr 21, 2020
Grant date	Apr 21, 2020

Abstract

Described herein are methods for the efficient production of a heteromultimeric protein, such as a bispecific antibody. Heteromultimeric proteins may be capable of specifically binding to more than one target molecule or different epitopes on a single target molecule. The methods modulate parameters to improve assembly of the heteromultimeric proteins at higher yield and efficiency than otherwise possible. Also described are compositions comprising a hinge-containing polypeptide, such as a half-antibody.

First claim

What is claimed is:

1. A composition comprising an antibody moiety and a solubilizer selected from the group consisting of arginine, histidine, and sucrose, wherein the pH of the composition is between pH 5 to pH 9, wherein the antibody moiety consists of a half-antibody.

2. The composition of claim 1 , wherein the solubilizer is arginine or histidine.

3. The composition of claim 1 , wherein the solubilizer is an arginine salt or a histidine salt.

4. The composition of claim 1 , wherein the solubilizer is an arginine derivative or a histidine derivative.

5. The composition of claim 1 , wherein the solubilizer is arginine HCl or histidine HCl.

6. The composition of claim 2 , wherein the arginine or histidine is present at a concentration between 20 mM and 1 M.

7. The composition of claim 6 , wherein the arginine or histidine is present at a concentration between 20 mM and 400 mM.

8. The composition of claim 1 , wherein the composition comprises both arginine and histidine.

9. The composition of claim 8 , wherein the arginine and histidine are each present at a concentration between 20 mM and 200 mM or between 50 mM and 200 mM.

10. The composition of claim 8 , wherein the arginine is present at a concentration of 50 mM and the histidine is present at a concentration of 200 mM.

11. The composition of claim 1 , wherein the half-antibody is an IgG half-antibody.

12. The composition of claim 11 , wherein the IgG half-antibody is of the IgG1, IgG2 or IgG4 isotype.

13. The composition of claim 1 , wherein the half-antibody comprises an Fc component.

14. The composition of claim 1 , wherein the half-antibody comprises a VL domain, a VH domain, a hinge domain, a CH 2 domain and a CH 3 domain.

15. The composition of claim 14 , wherein the half-antibody comprises a single chain polypeptide and wherein said single chain polypeptide comprises domains positioned relative to each other in an N-terminal to C-terminal direction as follows: V L -tether-V H -hinge-CH 2 —CH 3 .

16. The composition of claim 14 , wherein the half-antibody further comprises a C L domain and a CH 1 domain.

17. The composition of claim 16 , wherein the half-antibody comprises a single chain polypeptide and wherein said single chain polypeptide comprises domains positioned relative to each other in an N-terminal to C-terminal direction as follows: V L -C L -tether-V H -CH 1 -hinge-CH 2 —CH 3 .

18. The composition of claim 1 , wherein the half-antibody comprises a knob or a hole.

19. The composition of claim 1 , further comprising a second half-antibody.

20. The composition of claim 19 , wherein the first half-antibody comprises a knob and the second half-antibody comprises a hole.

21. The composition of claim 1 , wherein the pH of the composition is pH 6 to pH 9.

22. The composition of claim 21 , wherein the pH of the composition is pH 7 to pH 9.

23. The composition of claim 18 , wherein the knob comprises a T366W substitution, and the hole comprises T366S, L368A, and Y407V substitutions.

24. The composition of claim 20 , wherein the knob comprises a T366W substitution, and the hole comprises T366S, L368A, and Y407V substitutions.

25. The composition of claim 1 , further comprising a weak reductant.

26. The composition of claim 25 , wherein the weak reductant is glutathione (GSH).

27. The composition of claim 26 , wherein GSH is present in 2-600× molar excess with respect to the total amount of the half-antibody.

Assignees

• Genentech Inc

Inventors

• Giese Glen
• Lim Amy
• Persson Josefine
• Scheer Justin
• Williams Ambrose

Classifications

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title

• C07K2317/53

  Hinge · CPC title

• C07K2317/524

  CH2 domain · CPC title

• C07K2317/522

  CH1 domain · CPC title

• C07K2317/31

  multispecific · CPC title