Preparation and identification of monoclonal antibodies against VP22 protein of duck enteritis virus.

In this study,the UL49 gene was amplified from the genome of duck enteritis virus(DEV) Clone-03 strain by PCR method and was cloned into pMD-18 vector.UL49 gene was subcloned into the prokaryotic expression vector pET-30a.The E.coli BL21(DE3) contained the recombinant plasmid was induced by IPTG and the recombinant protein was analyzed by SDS-PAGE.The recombinant protein with a molecular weight of 36 ku could be detected by western blot using the antiserum against DEV.Monoclonal antibodies(MAbs) against VP22 protein of DEV were developed using recombinant protein VP22 as antigen.Four MAbs against VP22 were obtained following screening by indirect enzyme-linked immunosorbent assay(ELISA),western blot and indirect immunofluorescence assay.Four MAbs,designated as 2B10,2G1,3F10 and 3G2 respectively,could recognize the VP22 protein in DEV-infected chicken embryo fibroblasts.This study paved the way for future study of biological function and antigenic epitope of VP22 protein.