title: Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein.
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.
authors.affiliation: State Key Lab of State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin, 150001, China.
authors.forename: Xiuchen
authors.initials: X
authors.lastname: Yin
authors.forename: Shumei
authors.initials: S
authors.lastname: Zhang
authors.forename: Youlan
authors.initials: Y
authors.lastname: Gao
authors.forename: Jinzhe
authors.initials: J
authors.lastname: Li
authors.forename: Shuyi
authors.initials: S
authors.lastname: Tan
authors.forename: Hongyu
authors.initials: H
authors.lastname: Liu
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Wu
authors.forename: Yuhuan
authors.initials: Y
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Liu
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology*; D000229:Dependovirus / Q000276:immunology*; D004372:Ducks / Q000821:virology*; D005455:Fluorescent Antibody Technique; D005777:Geese / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; C442558:VP3 protein, Dependovirus