Bivalent, bispecific antibodies

• US 10,138,293 B2
• Filed: 01/31/2012
• Issued: 11/27/2018
• Est. Priority Date: 12/21/2007
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Klein, Christian, Schaefer, Wolfgang
• Primary Examiner(s): Mertz, Prema M
• Application Number:
  US13/362,000
• Publication Number:
  US 20120225071A1
• Time in Patent Office: 2,492 Days
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention relates to nucleic acids which encode the heavy chains and light chains of a novel domain exchanged, bivalent, bispecific antibody, and vectors comprising the same.

Claims

1. A bivalent, bispecific antibody comprising two pairs of light chains and heavy chains, each light chain comprising a variable region and a constant region, each heavy chain comprising a variable region and a constant region, wherein:

a) the light chain and heavy chain of the first of the two pairs specifically bind to a first antigen, wherein the light chain comprises the following domains in N-terminal to C-terminal direction VL, CL and the heavy chain comprises the following domains in N-terminal to C-terminal direction VH, CH1, CH2, CH3; and

b) the light chain and heavy chain of the second of the two pairs specifically bind to a second antigen, wherein the light chain comprises the following domains in N-terminal to C-terminal direction VH, CL and the heavy chain comprises the following domains in N-terminal to C-terminal direction VL, CH1, CH2, CH3.

2. The bivalent, bispecific antibody of claim 1, wherein the CH3 domain of one heavy chain is altered so that within the interface the CH3 domain of one heavy chain that meets the interface of the CH3 domain of the other heavy chain within the bivalent, bispecific antibody, an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the interface of the CH3 domain of one heavy chain which is positionable in a cavity within the interface of the CH3 domain of the other heavy chain, and

wherein the CH3 domain of the other heavy chain is altered, so that within the interface the CH3 domain of one heavy chain that meets the interface of the CH3 domain of the other heavy chain within the bivalent, bispecific antibody, an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the interface of the CH3 domain of the other heavy chain within which a protuberance within the interface of the CH3 domain of the one heavy chain is positionable.

3. The antibody according to claim 2, wherein the amino acid residue having a larger side chain volume is selected from the group consisting of arginine (R), phenylalanine (F), tyrosine (Y), tryptophan (W).

4. The antibody according to claim 2, wherein the amino acid residue having a smaller side chain volume is selected from the group consisting of alanine (A), serine (S), threonine (T), valine (V).

5. The antibody according to claim 2, wherein both CH3 domains are further altered by the introduction of cysteine (C) as amino acid in the corresponding positions of each CH3 domain.

6. The antibody according to claim 3, wherein both CH3 domains are further altered by the introduction of cysteine (C) as amino acid in the corresponding positions of each CH3 domain.

7. The antibody according to claim 4, wherein both CH3 domains are further altered by the introduction of cysteine (C) as amino acid in the corresponding positions of each CH3 domain.

8. A method for the preparation of a bivalent, bispecific antibody according to claim 1 comprising the steps of

a) transforming a host cell with

vectors comprising nucleic acid molecules encoding the light chain and heavy chain of the first of the two pairs specifically binding to a first antigen;

vectors comprising nucleic acid molecules encoding the light chain and heavy chain of the second of the two pairs specifically binding to a second antigen;

b) culturing the host cell under conditions that allow synthesis of said antibody molecule; and

c) recovering said antibody molecule from said culture.

9. A composition comprising a bivalent, bispecific antibody according to claim 1 and at least one pharmaceutically acceptable excipient.

10. A composition comprising a bivalent, bispecific antibody according to claim 3 and at least one pharmaceutically acceptable excipient.

11. A composition comprising a bivalent, bispecific antibody according to claim 5 and at least one pharmaceutically acceptable excipient.

12. A composition comprising a bivalent, bispecific antibody according to claim 7 and at least one pharmaceutically acceptable excipient.