Disulfide stabilized foldon polypeptides

Patent metadata

Field	Value
Publication number	US-9550813-B2
Application number	US-201314401754-A
Country	US
Kind code	B2
Filing date	May 22, 2013
Priority date	May 22, 2012
Publication date	Jan 24, 2017
Grant date	Jan 24, 2017

Abstract

Intermolecular disulfide stabilized foldon polypeptides are provided.

First claim

What is claimed is:

1. A cysteine-substituted foldon polypeptide, wherein two amino acids of a native foldon polypeptide are substituted with cysteine residues that can form intermolecular disulfide bonds when the foldon is in a trimeric quaternary structure, wherein the native foldon polypeptide has the amino acid sequence set forth in SEQ ID NO:1 and wherein the amino acid substitutions are selected from [G10C, D17C]; [R8C, D17C]; [R8C, G18C]; [P7C, G18C]; [A12C, K16C]; and [Y13C, R15C].

2. The cysteine-substituted foldon polypeptide of claim 1 , wherein the amino acid substitutions are selected from [A12C, K16C]; [R8C, D17C]; and [R8C, G18C].

3. The cysteine-substituted foldon polypeptide of claim 1 , wherein the cysteine-substituted foldon polypeptide is a monomer.

4. The cysteine-substituted foldon polypeptide of claim 1 , wherein the cysteine-substituted foldon polypeptide is a trimer.

5. The cysteine-substituted foldon polypeptide of claim 1 , wherein the cysteine-substituted foldon polypeptide is joined to a fusion partner.

6. The cysteine-substituted foldon polypeptide of claim 5 , wherein the fusion partner is a viral antigen.

7. The cysteine-substituted foldon polypeptide of claim 1 , further comprising one or more of a tag for purification, an unnatural amino acid, and site for protease cleavage.

8. The cysteine-substituted foldon polypeptide of claim 7 , wherein the cysteine-substituted foldon polypeptide is a fusion protein, and comprises at least one unnatural amino acid at a defined site.

9. The cysteine-substituted foldon polypeptide of claim 8 , where the cysteine-substituted foldon polypeptide is a trimer, and the unnatural amino acid is used to covalently join one chain of the trimer to a virus like particle.

10. A method of producing a cysteine-substituted foldon polypeptide according to claim 1 , comprising introducing a polynucleotide sequence encoding the cysteine-substituted foldon polypeptide operably linked to a promoter, into a cell or cell free protein synthesis reaction mix and incubating for a period of time sufficient to synthesize the protein.

Assignees

• Univ Leland Stanford Junior

Inventors

• Lu Yuan
• Swartz James R

Classifications

• C07K2319/50

  containing protease site · CPC title

• C12N2760/16022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C07K2319/35

  containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin · CPC title

• C12N2795/10122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C07K2319/22

  containing a Strep-tag · CPC title