Novel Norovirus Particle for Use as an Antiviral or Vaccine

• US 20090280139A1
• Filed: 06/15/2006
• Published: 11/12/2009
• Est. Priority Date: 06/16/2005
• Status: Active Grant

• Current Assignee: Children's Hospital & Medical Center
• Original Assignee: Children's Hospital & Medical Center
• Inventors: Tan, Ming, Jiang, Xi
• Granted Patent: US 8,277,819 B2
• Time in Patent Office: Days
• US Class Current: 424/186.100
• CPC Class Codes: A61K 2039/5258 Virus-like particles
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2770/16022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/16023 Virus like particles [VLP]
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

Norovirus capsid protein monomers having only the P domain, and not the hinge or S domain, can assemble spontaneously into an icosahedral form herein called the P-particle. Factors affecting the formation and stability of the P-particle, as well as providing methods for diagnosing and treating Norovirus-infected individuals and creating a vaccine for prevention of Norovirus infection are presented.

Claims

1-2. -2. (canceled)

3. A kit for diagnosing a norovirus infection in an individual, the kit comprising:

(a) purified recombinant P-particles from a Norovirus strain; and

(b) an enzyme-linked immuno assay that employs the purified recombinant P-particles to detect antibodies to the Norovirus strain in the individual'"'"'s serum.

4. The kit of claim 3, wherein the Norovirus strain is selected from the group consisting of VA387, MOH and NV.

5. The kit of claim 3, wherein use of the enzyme-linked immuno assay includes the steps of:

(1) pre-coating the purified recombinant P-particles onto a plate used in the enzyme-linked immuno assay;

(2) adding the individual'"'"'s serum to the plate;

(3) adding anti-human immunoglobulin which is coupled to an enzyme to the plate;

(4) adding a substrate to the plate which changes color when cleaved by the enzyme; and

(5) determining if the substrate changes color, whereby a positive diagnosis of Norovirus infection can be made.

9. The kit of claim 3, wherein the P-particle is a purified recombinant P-particle in the shape of an icosahedron, the icosahedron comprised of 24 monomer units of SEQ ID NO:

2.

10. The kit of claim 3, wherein the P-particle is a purified recombinant P-particle from a Norovirus strain selected from the group consisting of VA387, MOH and NV.

6-8. -8. (canceled)

11. A vaccine for the prevention of Norovirus infection in humans, the vaccine comprising a therapeutic amount of a compound having increased receptor binding activity with human histo-blood group antigens in a suitable sterile, nontoxic physiological medium.

12. The vaccine of claim 11, wherein the compound having increased receptor binding activity with human histo-blood group antigens is a P-particle in a suitable sterile, nontoxic physiological medium.

13. The vaccine of claim 12, wherein the P-particle comprises capsid protein monomers from the norovirus strain VA387.

14. The vaccine of claim 13, wherein the capsid protein monomers are comprised of protein selected from the group consisting of SEQ ID NO:

1 SEQ ID NO;

2, a P protein, a dimer of the P protein, a pentamer of dimers of the P protein, and mixtures thereof.

15-16. -16. (canceled)

17. A method of inducing an immune response in an individual against Norovirus, the method comprising the step of orally or parenterally administering to an individual at least one immunologically effective dose of a composition comprising an immunogen recombinantly expressed or synthesized from a cDNA comprising:

a fragment of the cDNA sequence of the capsid protein of norovirus strain VA387 and a pharmaceutical carrier, wherein the dose is effective in inducing the immune response in the individual.

18. The method of claim 17, wherein the fragment is selected from the group consisting of SEQ ID NO:

1 and SEQ ID NO;

2.

19. The method of claim 17, wherein the immunogen recombinantly expressed or synthesized from cDNA is an isolated P-particle from a Norovirus strain.

20. The method of claim 17, wherein the isolated P-particle comprises a plurality of P-Particles which have been purified and which have been cysteine-modified.

21. The method of of claim 20, wherein the cysteine has been linked to the end of the P1 subdomain, the resulting P-particle being more stable than the original P-particle.