Expression, purification and glycosylation analysis of chicken infectious bursal disease virus VP2 in yeast

ABSTRACT

Infectious bursal disease [IBD], a highly contagious and devastating disease in young chicken, is caused by infectious bursal disease virus [IBDV]. To improve the immunogenicity of recombinant IBDV subunit vaccine, an attempt was made to find a new way to prepare IBD vaccine containing glycosylated mVP[2] antigen. Firstly, IBDV mVP[2] gene [with a nucleic acid sequence encoding B cell epitope of IBDV [KFDQML] in the 5'-end of the VP[2], with a nucleic acid sequence encoding B cell epitope of IBDV [LASP] and [His] 6-tag in the 3'-end of the VP[2]] was cloned. Secondly, IBDV mVP[2] protein was expressed in the methylotrophic yeast Pichia pastoris which can secret glycosylated protein. The recombinant mVP[2] protein could be stained pink with periodic acid-schiff reagents [PAS], which showed that mVP[2] was glycosylated. Finally, IBDV mVP[2] protein was purified with His-Trap [1 mL] affinity chromatography. These results indicate that glycosylated IBDV VP[2] protein modified with epitope peptides can be expressed in Pichia pastoris, which lay the groundwork for the development of a recombinant infectious bursal disease vaccine with high immunogenicity