Prokaryotic expression and development of monoclonal antibody against glycoprotein 2(GP2)of PRRSV CH-1a strain

The ORF2 gene of PRRSV strain CH-la was truncated and cloned into a prokaryotic expression vector pGEX-6p-1,and a fusion-expressed protein rtGP2 of 40 Ku was obtained in E.coil.The protein rtGP2 showed a strong reaction to the PRRSV-positive sera in Western-blotting.BALB/c mice were immunized intraperitoneally with purified rtGP2 protein. Murine myeloma cells were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA coated with rtGP2 and GST was used to screen hybridomas for production of specific antibody in hybridoma culture fluid.One hybridomas clone which producedMAb steadily was obtained,named 26D8.The McAb showed strong reactivity in IFA on the PRRSV infected Marc-145 cells.The McAb is belonging to IgG1 isotype,and the light chain of the antibody isκchain,which was isotyped by ELISA using a commercial kit.The McAb against PRRSV GP2 protein could be used for further analysis of the structure and function of PRRSV.