Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

Abstract

Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the major capsid protein VP2 of PPV was amplified using viral nucleic acid extract from swine serum and inserted into yeast Saccharomyces cerevisiae expression plasmid. Electron microscopy analysis revealed that virus-like particle composed of major capsid protein VP2 of PPV was synthesised that further was used to immunize mice. The specificity of nine newly generated MAbs was proven by immunofluorescence analysis of PPV-infected cells. Recombinant antigen based indirect ELISA to detect PPVspecific antibodies in swine sera was developed and evaluated. The sensitivity and specificity of the new assay were found to be 89.8% and 100% respectively. In conclusion, yeast S. cerevisiae offers an alternative of expression system for generating recombinant PPV VP2 protein VLPs and PPV-specific MAbs of diagnostic relevance.