Method for producing yeast expressed hpv types 6 and 16 capsid proteins

Abstract

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 diploid cell comprising vectors for expressing capsid proteins from at least two Human Papilloma Viruses (HPV), wherein said at least two HPVs are HPV type 6 and HPV type 16, wherein the capsid proteins comprise a late 1 (L1) capsid protein from HPV type 6, an L1 capsid protein from HPV type 16, a minor capsid protein late 2 (L2) from HPV type 6 and an L2 capsid protein from HPV type 16. 
     
     
         2 . The diploid cell of  claim 1 , wherein the cell is a yeast cell. 
     
     
         3 . The diploid cell of  claim 2 , wherein the yeast is  Saccharomyces cerevisiae.    
     
     
         4 . A method for producing a diploid cell according to  claim 1  comprising:
 a) cloning said capsid proteins into expression cassettes comprising the same promoters and termination sequences; and 
 b) expressing said cassettes in the same diploid cell. 
 
     
     
         5 . The method of  claim 4 , wherein the diploid cell is a yeast cell. 
     
     
         6 . The method of  claim 5 , wherein the yeast is  Saccharomyces cerevisiae.    
     
     
         7 . The method of  claim 4 , wherein the L1 expression cassettes are cloned into non-integrative vectors, and the L2 expression cassettes are cloned into integrative vectors. 
     
     
         8 . The method of  claim 7  wherein the non-integrative vector is pBS24.1. 
     
     
         9 . The method of  claim 7  wherein the integrative vector is pUC8. 
     
     
         10 . A method for producing a virus-like particle (VLP) comprising:
 (a) cloning capsid proteins that comprise a late 1 (L1) capsid protein from HPV type 6, an L1 capsid protein from HPV type 16, a minor capsid protein late 2 (L2) from HPV type 6 and an L2 capsid protein from HPV type 16 into expression cassettes comprising the same promoters and termination sequences; and   (b) expressing the cassettes in the same diploid cell.   
     
     
         11 . The method of  claim 10  wherein the diploid cell is a yeast cell. 
     
     
         12 . The method of  claim 11  wherein the yeast is  Saccharomyces cerevisiae.    
     
     
         13 . The method of  claim 10  wherein said L1 expression cassettes are cloned into non-integrative vectors, and said L2 expression cassettes are cloned into integrative vectors. 
     
     
         14 . The method of  claim 13  wherein the non-integrative vector is pBS24.1. 
     
     
         15 . The method of  claim 13  wherein the integrative vector is pUC8.