Preparation And Identification Of Gp5 Protein To Develop Monoclonal Antibody Against Porcine Reproductive And Respiratory Syndrome Virus

The aim of this paper was for obtaining monoclonal antibodies to against Porcine Reproductive and Respiratory Syndrome Virus ( PRRSV), recombined GP5 protein as an antigen were be used to immunilized BALB/C mice. Three monoclonal antibodies were be selected by indirect ELISA and many of property were identificated.Among the three fusion cell strains, 8C9 belonged to IgG1 subclass of Ig, 4B4 and 5H2 belonged to IgG2a subclass of Ig. And the number of fusion cell chromatosome under 102 to 108 items; The results of specificity test indicated that the two cell strains monoclonal antibodies were specific reaction for PRRSV and GP5 protein, one for GST, all of them with no reaction to CSFV and PCV. Their titers in cell culture supernatant were reach to 1:104, The titers of abdomen liquor were1: 105, respectively.The purified protein of monoclonal antibodies by SDS-PAGE showed the molecular weights of the heavy chain and light chain were about 45.0 kD and 25.0 kD, respectively. By stratum super stratum test, the antigenic epitopes of 8C9 and 4B4 were not coincidence, this result indicated that the epitopes recognized 8C9 is different to 4B4.In neutralization activities test, the setting virus control, mAb control and cell control were normal until the time end, the result showed that the prepared 4B4 mAb can protected 50% cells no CPE in the dilution reach to 1:256, but 8C9 have no any neutralization activities to PRRSV.