Truncated L1 protein of human papillomavirus type 33

Patent metadata

Field	Value
Publication number	US-9249193-B2
Application number	US-201214122150-A
Country	US
Kind code	B2
Filing date	May 22, 2012
Priority date	May 25, 2011
Publication date	Feb 2, 2016
Grant date	Feb 2, 2016

Abstract

The present invention relates to a truncated L1 protein of Human Papillomavirus (HPV) Type 33, a sequence encoding the same, a method for preparing the same, and a virus-like particle comprising the same, wherein the protein and the virus-like particle are useful for preventing HPV (particularly HPV33) infection, and a disease caused by HPV (particularly HPV33) infection, such as cervical cancer. The invention also relates to the use of the protein and the virus-like particle in the preparation of a pharmaceutical composition or a vaccine for preventing HPV (particularly HPV33) infection, and a disease caused by HPV (particularly HPV33) infection, such as cervical cancer.

First claim

The invention claimed is:

1. A truncated HPV33 L1 protein, wherein said truncated HPV33 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7.

2. An isolated nucleic acid, encoding the truncated HPV33 L1 protein according to claim 1 .

3. A vector comprising the isolated nucleic acid according to claim 2 .

4. An isolated host cell comprising the isolated nucleic acid according to claim 2 , or a vector comprising the isolated nucleic acid.

5. A HPV33 virus-like particle, comprising or consisting of the truncated HPV33 L1 protein according to claim 1 .

6. A composition comprising the truncated HPV33 L1 protein according to claim 1 , or an isolated nucleic acid encoding the truncated HPV33 L1 protein, or a vector comprising an isolated nucleic acid encoding the truncated HPV33 L1 protein.

7. A pharmaceutical composition or vaccine comprising the HPV33 virus-like particle according to claim 5 , and optionally, a pharmaceutically acceptable carrier, excipient or combination thereof.

8. A method for obtaining a truncated HPV33 L1 protein, comprising expressing the truncated HPV33 L1 protein of claim 1 with an E. coli expression system, and carrying out a purification treatment on the lysed supernatant containing said protein.

9. A method for preparing a vaccine, comprising blending the HPV33 virus-like particle according to claim 5 with a pharmaceutically acceptable carrier, excipient or combination thereof.

10. A method for inhibiting HPV33 infection or preventing a disease caused by HPV33 infection, comprising administering to a subject a prophylactically effective amount of the HPV33 virus-like particle according to claim 5 or a pharmaceutical composition or vaccine comprising the HPV33 virus-like particle and, optionally, pharmaceutically acceptable carriers and/or excipients.

11. A truncated HPV33 L1 protein according to claim 1 , wherein said truncated HPV33 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 4.

12. A composition comprising the host cell according to claim 4 .

13. A composition comprising the HPV33 virus-like particle according to claim 5 .

14. A pharmaceutical composition or vaccine according to claim 7 , wherein the HPV33 virus-like particle is present in an amount effective for inhibiting HPV33 infection or preventing cervical cancer caused by HPV33 infection.

15. A method for obtaining a truncated HPV33 L1 protein according to claim 8 , wherein the method comprises the steps of:
a) expressing the truncated protein in E. coli;
b) disrupting the E. coli , which has expressed the truncated protein, in a solution with a salt concentration of 100 mM to 600 mM, and isolating the supernatant;
c) decreasing the salt concentration of the supernatant obtained in step b) to 100 mM or less by using water or a solution at a low salt concentration, and collecting a precipitate; and
d) re-dissolving the precipitate obtained in step c) in a solution with a salt concentration of 150 mM to 2500 mM and adding a reductant to the solution, and then isolating the resultant solution, wherein the resultant solution contains the truncated HPV33 L1 protein with a purity of at least 50%.

16. A method for preparing a HPV33 virus-like particle comprising or consisting of a truncated HPV33 L1 protein, wherein said truncated HPV33 L1 protein has an amino acid sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7, the method comprising the steps of:
a) purifying the truncated HPV33 L1 protein as obtained by the method of claim 15 through a chromatography; and
b) removing the reductant from the truncated protein obtained in step a).

17. A method for inhibiting HPV33 infection or preventing a disease caused by HPV33 infection, comprising administrating to a subject a prophylactically effective amount of the HPV33 virus-like particle obtained by the method according to claim 16 .

18. A method for inhibiting HPV33 infection or preventing a disease caused by HPV33 infection, comprising administrating to a subject a prophylactically effective amount of the vaccine obtained by the method according to claim 9 .

19. A method for inhibiting HPV33 infection or preventing a disease caused by HPV33 infection according to claim 10 , wherein the disease caused by HPV33 infection is cervical cancer.

Assignees

• Li Shaowei
• Kong Xianglin
• Wei Minxi
• Pan Huirong
• Zhang Jun
• Xia Ningshao
• Univ Xiamen
• Xiamen Innovax Biotech Co Ltd

Inventors

• Li Shaowei
• Kong Xianglin
• Wei Minxi
• Pan Huirong
• Zhang Jun
• Xia Ningshao

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• A61K39/12

  Viral antigens · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/20

  for DNA viruses · CPC title

• C12N2710/20023

  Virus like particles [VLP] · CPC title