Method for preparing virus-like particle and recombinant baculovirus used therein

Abstract

A recombinant baculovirus is provided for preparing picornavirus virus-like particles (VLP), wherein Chitinase A (ChiA) and Cathepsin V (v-cath) genes of the recombinant baculovirus are functionally disrupted and the recombinant baculovirus includes a picornavirus capsid protein gene under control of a strong promoter, and includes a protease gene configured for encoding a protease for hydrolyzing the capsid protein under control of a weak promoter. The recombinant baculovirus of the present invention may adopt High Five or Sf-9 cells for manufacturing enterovirus virus-like particles with improved stability and higher yields in comparison with the conventional arts. A method for preparing virus like particles is also herein provided.

What is claimed is: 
     
         1 . A recombinant baculovirus comprising:
 a first nucleotide sequence, for being translated to a capsid protein of a picornavirus;   a first promoter, located in the upstream region of the first nucleotide sequence;   a second nucleotide sequence, for being translated to a protease of the picornavirus; and   a second promoter, located in the upstream region of the second nucleotide sequence, wherein the first promoter is a strong promoter and the second promoter is a weak promoter in the context of recombinant baculovirus infection to an insect cell and ChiA and v-cath genes of the recombinant baculovirus are functionally disrupted.   
     
     
         2 . The recombinant baculovirus according to  claim 1 , wherein the first promoter comprises a baculovirus polyhedrin promoter or a baculovirus P10 promoter. 
     
     
         3 . The recombinant baculovirus according to  claim 1 , wherein the second promoter comprises a baculovirus immediate early promoter, a cytomegalovirus immediate early gene (CMV-1B) promoter or a  Drosophila  hsp70 promoter. 
     
     
         4 . The recombinant baculovirus according to  claim 1 , wherein the second promoter comprises a fragment of the strong promoter. 
     
     
         5 . The recombinant baculovirus according to  claim 1 , wherein the insect cell comprises a Sf-9 cell strain or a High Five™ cell strain. 
     
     
         6 . The recombinant baculovirus according to  claim 1 , wherein the picornavirus comprises an enterovirus. 
     
     
         7 . The recombinant baculovirus according to  claim 6 , wherein the enterovirus comprises enteroviruses 68-71, Coxsackie A viruses or Coxsackie B viruses. 
     
     
         8 . The recombinant baculovirus according to  claim 1 , wherein the recombinant baculovirus is used for preparing a vaccine.