Engineered heterodimeric protein domains

Patent metadata

Field	Value
Publication number	US-9505848-B2
Application number	US-201414505653-A
Country	US
Kind code	B2
Filing date	Oct 3, 2014
Priority date	Mar 24, 2006
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

The present invention provides an engineered multidomain protein including at least two nonidentical engineered domains, each of which contains a protein-protein interaction interface containing amino acid sequence segments derived from two or more existing homologous parent domains, thereby conferring on the engineered domains assembly specificities distinct from assembly specificities of the parent domains. In particular, the engineered domains form heterodimers with one another preferentially over forming homodimers. Methods of designing and using the engineered proteins are also included.

First claim

What is claimed is:

1. A nucleic acid encoding a heterodimeric fusion protein comprising first and second polypeptides, each comprising a CH3 domain mediating the heterodimerization of the first and second polypeptides, wherein each CH3 domain is a modified IgG CH3 domain substituted with corresponding amino acid residues from an IgA CH3 domain,
wherein each modified IgG CH3 domain comprises substitutions selected from the group consisting of: Glu at Kabat position 347, His at Kabat position 349, Pro at Kabat position 354, Leu at Kabat position 359, Asn at Kabat position 360, Leu at Kabat position 362, Thr at Kabat position 364, Arg at Kabat position 370, Lys at Kabat position 390, Leu at Kabat position 392, Trp at Kabat position 394, Ala at Kabat position 395, Arg at Kabat position 397, Glu at Kabat position 399, Pro at Kabat position 400, Ala at Kabat position 405, Thr at Kabat position 407, Ile at Kabat position 409, and Arg at Kabat position 411;
wherein the substitutions in the first and second polypeptides occur at different positions; and
wherein heterodimerization of the modified IgG CH3 domains is enhanced compared to IgG CH3 domains without the substitutions.

2. The nucleic acid of claim 1 , wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 10 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 11.

3. The nucleic acid of claim 1 , wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 3 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 6.

4. The nucleic acid of claim 1 , wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 5 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 8.

5. A cell comprising the nucleic acid of claim 1 .

6. A nucleic acid encoding a heterodimeric fusion protein comprising first and second polypeptides, each comprising a CH3 domain mediating the heterodimerization of the first and second polypeptides, wherein each CH3 domain is a modified IgG CH3 domain substituted with corresponding amino acid residues from an IgA CH3 domain,
wherein each modified IgG CH3 domain comprises substitutions selected from the group consisting of: Glu at Kabat position 347, His at Kabat position 349, Pro at Kabat position 354, Leu at Kabat position 359, Asn at Kabat position 360, Leu at Kabat position 362, Thr at Kabat position 364, Arg at Kabat position 370, Lys at Kabat position 390, Leu at Kabat position 392, Trp at Kabat position 394, Ala at Kabat position 395, Arg at Kabat position 397, Glu at Kabat position 399, Pro at Kabat position 400, Thr at Kabat position 407, Ile at Kabat position 409, and Arg at Kabat position 411;
wherein the substitutions in the first and second polypeptides occur at different positions; and
wherein heterodimerization of the modified IgG CH3 domains is enhanced compared to IgG CH3 domains without the substitutions.

7. A cell comprising the nucleic acid of claim 6 .

Assignees

• Merck Patent Gmbh

Inventors

• Davis Jonathan H
• Huston James S

Classifications

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2317/524

  CH2 domain · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K2317/526

  CH3 domain · CPC title