title: Application of GP5 protein to develop monoclonal antibody against porcine reproductive and respiratory syndrome virus.
abstract: In this study, a panel of monoclonal antibodies (mAbs) against Porcine reproductive and respiratory syndrome virus(PRRSV), named as 8C9 and4B4, were produced by fusing SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with the PRRSV (TCID(50)=5.5), screened by the indirect ELISA and subjected to several limiting dilutions. mAbs were then identified by biological characterization. Among the two fusion cell strains, 8C9 belonged to the IgG1 subclass and 4B4 belonged to the IgG2a subclass. The titers in cell culture supernatant and abdomen liquor reached to 1:10(4)and 1:10(5), respectively. The specificity test indicated that the two cells had specific reactions for the PRRSV and GP5 protein respectively, and no reaction with Classical swine fever virus (CSFV) or Swine vesicular disease virus (SVDV). The molecular weights of the heavy chain and light chain were about 45.0 kDa and 25.0 kDa, respectively. In neutralization activity tests, the results showed that the prepared mAb 4B4 can protect 50% of cells with no CPE in dilution up to 1:512, but mAB 8C9 has no neutralization activities to PRRSV.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Tian
authors.forename: Yan
authors.initials: Y
authors.lastname: Cheng
authors.forename: Jin-yang
authors.initials: JY
authors.lastname: Wu
authors.forename: Jian-hui
authors.initials: JH
authors.lastname: He
authors.forename: You-jun
authors.initials: YJ
authors.lastname: Shang
authors.forename: Xiang-tao
authors.initials: XT
authors.lastname: Liu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D006692:Classical Swine Fever Virus / Q000276:immunology; D029822:Enterovirus B, Human / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology*; D013552:Swine; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D014759:Viral Envelope Proteins; C114789:glycoprotein 5, PRRSV