Protein complex system for increased immunogenicity and functionality, and methods making and use

Patent metadata

Field	Value
Publication number	US-9562077-B2
Application number	US-201313803057-A
Country	US
Kind code	B2
Filing date	Mar 14, 2013
Priority date	Jul 11, 2012
Publication date	Feb 7, 2017
Grant date	Feb 7, 2017

Abstract

Genes for proteins which spontaneously form dimers and/or oligomers can be recombinantly linked together, which upon expression in E. coli produces stable dimeric fusion proteins that spontaneously self-assemble into enormous, polyvalent complexes having increased immunogenicity and functionality. Linear, network and agglomerate complexes with enormous sizes and polyvalences are constructed using glutathione S-transferase, Norovirus P domains (NoV P − and NoV + ), the protruding (P) domain of hepatitis E virus (HEV P), the astrovirus P domain (AstV), a monomeric peptide epitope (M2e of influenza virus), and/or a protein antigen (VP8* of rotavirus) fused in different combinations. The resulting complexes can contain hundreds to thousands NoV P-protein, HEV, AstV, M2e and/or VP8* copies and exhibit higher immunogenicity than the individual proteins alone. The large size and multivalent nature of the complexes are candidates as a bivalent or multivalent vaccines against Norovirus and other pathogens, and for generation of antibodies for diagnosis and research purposes.

First claim

What is claimed is:

1. A vaccine or vaccine platform having a linear network structure comprising two dimeric proteins derived from different species,
wherein said two dimeric proteins are connected by a linker peptide;
wherein said two dimeric proteins dimerize to form a homo-dimer;
wherein said linear network structure is formed via intermolecular interactions between said two dimeric proteins;
wherein at least one of the dimeric proteins comprises a P domain of a virus selected from the group consisting of Norovirus (NoV), Hepatitis E virus (REV), and Astrovirus (AstV).

2. The vaccine or vaccine platform according to claim 1 , wherein at least one dimeric protein is SEQ. ID. NO:17.

3. The vaccine or vaccine platform according to claim 1 , wherein at least one of the two dimeric proteins further comprises SEQ. ID. NO:20.

4. The vaccine or vaccine platform according to claim 1 , wherein the linker peptide comprises a length of amino acid units sufficient to allow the at least two dimeric proteins to fold into the proper structures without interference.

5. The vaccine or vaccine platform according to claim 1 , wherein at least one of the at least two dimeric protein proteins includes a foreign antigen.

6. The vaccine or vaccine platform according to claim 5 , wherein the foreign antigen comprises a foreign viral antigen.

7. The vaccine or vaccine platform according to claim 1 , further comprising a monomer of a dimeric protein, which forms a dimer cap at the same dimeric protein exposed on a dimeric fusion protein.

8. A vaccine or vaccine platform having a network protein complex structure comprising three dimeric proteins,
wherein said three dimeric proteins are derived from different species;
wherein said three dimeric proteins are connected by a linker peptide;
wherein said dimeric proteins form a homo-dimer;
wherein said network protein complex structure is formed via intermolecular interactions between said three dimeric protein domains;
wherein at least one of said three dimeric proteins are one or more of a virus P domain selected from Norovirus (NoV), Hepatitis E virus (REV), and Astrovirus (AstV).

9. The vaccine or vaccine platform according to claim 8 , wherein at least one dimeric protein is SEQ. ID. NO:17.

10. The vaccine or vaccine platform according to claim 8 , wherein at least one of the two dimeric proteins is SEQ. ID. NO:20.

11. The vaccine or vaccine platform according to claim 8 , wherein the linker peptide comprises a length of amino acid units sufficient to allow the at least two dimeric proteins to fold into the proper structures without interference.

12. The vaccine or vaccine platform according to claim 8 , wherein at least one of the at least two dimeric proteins includes a foreign antigen.

13. The vaccine or vaccine platform according to claim 8 , wherein the foreign antigen comprises a foreign viral antigen.

14. The vaccine or vaccine platform according to claim 8 , further comprising a monomer of a dimeric protein, which forms a dimer cap at the same dimeric protein domain exposed on a dimeric fusion protein.

Assignees

• Children'S Hospital Medical Center

Inventors

• Tan Ming
• Jiang Xi
• Wang Leyi

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• C07K2319/70

  containing domain for protein-protein interaction · CPC title

• C12N2760/16022

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

• C12N2770/16022

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

• C07K2319/40

  containing a tag for immunodetection, or an epitope for immunisation · CPC title