Antibodies with engineered CH2 domains, compositions thereof and methods of using the same

Patent metadata

Field	Value
Publication number	US-12398197-B2
Application number	US-202117387223-A
Country	US
Kind code	B2
Filing date	Jul 28, 2021
Priority date	Jun 15, 2016
Publication date	Aug 26, 2025
Grant date	Aug 26, 2025

Abstract

The present disclosure relates to antibodies and antibody conjugates having one or more site-specific mutations in the CH2 domain of the heavy chain. The antibody variants disclosed herein can have improved characteristics (e.g., thermal stability, antibody yields, antibody titers, cell-killing) relative to a parent or wild type antibody, including aglycosylated parent or wild type antibodies. Pharmaceutical compositions, diagnostic compositions and kits comprising the same, as well as methods of using these compositions and kits for therapeutic and diagnostic purposes, are also described.

First claim

What is claimed:

1. A method for producing an antibody conjugate, comprising:
conjugating an antibody of the IgG class to one or more therapeutic or labeling moieties, wherein the antibody comprises:
(a) at least one amino acid substitution in the CH2 domain of the heavy chain of the antibody, and wherein the at least one amino acid substitution is selected from the group consisting of:
V264S;
V264S and V303R;
V264S and V305R; and
V264S, V303R, and V305R,
according to the EU numbering scheme; and

(b) at least one non-natural amino acid substitution, wherein the at least one non-natural amino acid substitution comprises a moiety selected from the group consisting of amino, carboxy, acetyl, hydrazino, hydrazido, semicarbazido, sulfanyl, azido, alkynyl, and tetrazine.

2. The method of claim 1 , further comprising substituting an amino acid at position F241 and/or F243 of the CH2 domain.

3. The method of claim 1 , wherein the antibody is aglycosylated.

4. The method of claim 1 , wherein the antibody is a bi-specific antibody.

5. The method of claim 1 , wherein the antibody is of a subclass selected from the group consisting of IgG1, IgG2, and IgG4.

6. The method of claim 1 , wherein the at least one non-natural amino acid is para-azido phenylalanine or para-azido methyl phenylalanine.

7. The method of claim 1 , wherein the antibody further comprises at least a second non-natural amino acid, and wherein the at least one non-natural amino acid residue comprises an acetyl moiety and the at least second non-natural amino acid residue comprises an azide moiety.

8. The method of claim 1 , wherein at least one of said therapeutic moieties or labeling moieties is linked to said antibody via a residue of the non-natural amino acid comprising an azide moiety.

9. The method of claim 1 , wherein at least one of said therapeutic moieties or labeling moieties is linked to said antibody via a residue of the non-natural amino acid comprising an acetyl moiety.

10. The method of claim 1 , wherein at least one of said therapeutic moieties or labeling moieties is linked to said antibody via a residue of the non-natural amino acid comprising an azide moiety and at least one of said therapeutic moieties or labeling moieties is linked to said antibody via a residue of the non-natural amino acid comprising an acetyl moiety.

11. The method of claim 1 , wherein said antibody is linked to said one or more therapeutic moieties or labeling moieties via one or more linkers.

12. The method of claim 1 , wherein said antibody is substantially pure.

13. The method of claim 1 , wherein said antibody is at least 95% by mass of the total antibody concentration.

14. The method of claim 1 , further comprising adding a pharmaceutically acceptable carrier.

15. The method of claim 1 , wherein the antibody is produced in a cell-free system.

16. The method of claim 15 , wherein the cell-free system utilizes a cell-free extract from E. coli.

Assignees

• Sutro Biopharma Inc

Inventors

• Calarese Daniel
• Yin Gang

Classifications

• C12N2015/8518

  expressing industrially exogenous proteins, e.g. for pharmaceutical use, human insulin, blood factors, immunoglobulins, pseudoparticles · CPC title

• C12N15/85

  for animal cells · CPC title

• C07K2317/522

  CH1 domain · CPC title

• C07K2317/41

  Glycosylation, sialylation, or fucosylation · CPC title

• C07K16/18

  against material from animals or humans · CPC title