Method for producing yeast expressed HPV types 6 and 16 capsid proteins
Mosaic VLPs of viral capsid proteins from different virus types are described, as are methods of making the same. Specifically, a diploid yeast strain that coexpresses the L1 and L2 capsid proteins of both HPV-6 and HPV-16 as mosaic VLPs is described. The mosaic VLPs induced the production of conformational antibodies against both L1 proteins upon administration to mice.
1. A virus-like particle (VLP) comprising capsid proteins from at least two different types of Human Papilloma Virus (HPV), wherein said capsid proteins comprise a first late 1 (L1) capsid protein from one type of HPV virus and a second L1 capsid protein from a second type of HPV virus.
2. The VLP of claim 1 , wherein the capsid proteins comprise the minor capsid protein late 2 (L2) from the different types of HPV.
3. A composition comprising the VLP of claim 1 and an adjuvant.
4. A composition comprising the VLP of claim 2 and an adjuvant.
5. The composition of claim 3 , wherein the adjuvant is MF59.
6. The composition of claim 4 , wherein the adjuvant is MF59.
7. A method for producing the VLP of claim 1 , said method comprising:
(a) cloning said capsid proteins into expression cassettes comprising the same promoters and termination sequences; and
(b) expressing said cassettes in the same host cell.
8. The method of claim 7 , wherein the host cell is a yeast cell.
9. The method of claim 8 , wherein the yeast is Saccharomyces cerevisiae.
10. The method of claim 7 , wherein the capsid proteins comprise the minor capsid protein late 2 (L2) from the different types of HPV and said L1 protein expression cassettes are cloned into non-integrative vectors, and said L2 proteins expression cassettes are cloned into integrative vectors.
11. The method of claim 10 , wherein the non-integrative vector is pBS24.1.
12. The method of claim 10 , wherein the integrative vector is pUC8.
13. A host cell comprising vectors for expressing capsid proteins from at least two different types of Human Papilloma Virus (HPV), wherein said capsid proteins comprise a first late 1 (L1) capsid protein from one type of HPV virus and a second L1 capsid protein from a second type of HPV virus.
14. The host cell of claim 13 , wherein said types of HPV are types 6 and 16.
15. The host cell of claim 14 , wherein said capsid proteins comprise the minor capsid protein late 2 (L2) from the different types of HPV.
16. The host cell of claim 13 , wherein said host cell is a diploid cell.
17. The host cell of claim 16 , wherein said host cell is yeast.
18. The host cell of claim 17 , wherein said yeast is Saccharomyces cerevisiae.
19. A method for inducing an immune response against more than one type of HPV virus comprising administering the VLP of claim 1 .
20. A method for inducing an immune response against more than one type of virus comprising administering the VLP of claim 2 .