Email updates

Keep up to date with the latest news and content from Virology Journal and BioMed Central.

Open Access Research

Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein

Xiuchen Yin1, Shumei Zhang1, Youlan Gao1, Jinzhe Li1, Shuyi Tan2, Hongyu Liu1, Xiaoying Wu1, Yuhuan Chen1, Ming Liu1* and Yun Zhang1*

Author Affiliations

1 State Key Lab of State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin, 150001, China

2 Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Haikou, 571100, China

For all author emails, please log on.

Virology Journal 2012, 9:288  doi:10.1186/1743-422X-9-288

Published: 23 November 2012

Abstract

Background

The VP3 protein of goose parvovirus (GPV) or Muscovy duck parvovirus (MDPV), a major structural protein, can induce neutralizing antibodies in geese and ducks, but monoclonal antibodies (MAbs) against VP3 protein has never been characterized.

Results

Three hybridoma cell lines secreting anti-GPV VP3 MAbs were obtained and designated 4A8, 4E2, and 2D5. Immunoglobulin subclass tests differentiated them as IgG2b (4A8 and 4E2) and IgG2a (2D5). Dot blotting assays showed that three MAbs reacted with His-VP3 protein in a conformation-independent manner. A competitive binding assay indicated that the MAbs delineated two epitopes, A and B of VP3. Immunofluorescence assay showed that MAbs 4A8, 4E2, and 2D5 could specifically bind to goose embryo fibroblast cells (GEF) or duck fibroblast cells (DEF) infected with GPV and MDPV. Dot blotting also showed that the MAbs recognized both nature GPV and MDPV antigen. Western blotting confirmed that the MAbs recognized VP3 proteins derived from purified GPV and MDPV particles. The MAbs 4A8 and 2D5 had universal reactivity to heterologous GPV and MDPV tested in an antigen-capture enzyme-linked immunosorbent assay.

Conclusions

Preparation and characterization of these the MAbs suggests that they may be useful for the development of a MAb-capture ELISA for rapid detection of both GPV and MDPV. Virus isolation and PCR are reliable for detecting GPV and MDPV infection, but these procedures are laborious, time-consuming, and requiring instruments. These diagnosis problems highlight the ongoing demand for rapid, reproducible, and automatic methods for the sensitive detection of both GPV and MDPV infection.

Keywords:
Monoclonal Antibody; GPV; MDPV; VP3