Development of monoclonal antibodies specific to the lineage 8 strains of porcine reproductive and respiratory syndrome virus
Porcine reproductive and respiratory syndrome virus(PRRSV),which is a pathogen that seriously endangers China's breeding industry,has multiple genotypes(lineages)and antigenic differences in circulating strains.In order to identify the circulating strains of dif-ferent lineages,the NSP2 recombinant protein was prepared by using the NSP2 hypervariable region gene fragment(tNSP2)of the PRRSV lineage 8 epidemic strain BB0907(a highly pathogenic strain),and the NSP2 recombinant protein obtained was used to immunize BALB/c mice.As a result,five monoclonal cell lines that could stably secrete antibodies were developed by cell fusion.The titers of the antibodies in ascites fluid in mice were ≥1∶216 000.The heavy chains of 3D6,6B2 and 6C8 were IgG1,the heavy chain of 8C2 was IgG2,while the heavy chain of 8H6 was IgG2a,and the light chains of the five McAbs were of kappa subtype.Four epitopes were identified by the construc-tion of NSP2 protein truncates,which were485GPLNFPTPSE494,555FPLAPSQNMG564,575EVLSEISDIL584 and 585NDTPAPVS593.These mono-clonal antibodies were observed to be able to react specifically with 8 strains of the PRRSV lineage by indirect immunofluorescence assay,but they did not react with the lineage 1 and lineage 5 strains.This study providedsignificantresearch materials for the differential diagnosis of and biological research onisolated PRRSV antigens.
porcine reproductive and respiratory syndrome virusnonstructural protein 2monoclonal antibodyB cell epitopes
万方数据
维普
