Preparation and identification of monoclonal antibodies against VP3 protein of goose parvovirus

The goose parvovirus(GPV) VP3 protein expressed in the recombinant E.coli Rosetta(DE3) was used as immunogen after purification by Ni-NTA.The 4 to 6 weeks-old BALB/c mice were intraperitoneally immunized with the VP3 protein for three times,then myeloma cells SP2/0 were fused with the spleen cells of the immunized BALB/c mice on day 3 post-last-immunization.Four hybridoma cell lines against the VP3 protein were obtained by screening with the indirect ELISA,named 1F1,2A9,3B11 and 4A2,respectively.The first monoclonal antibody(McAb) were identified to be IgG2b,and the others to be IgG1 with κ light chain.The ELISA titer of the 4 McAbs ascites were 1∶819 200,1∶1 638 400,1∶409 600 and 1∶819 200,respectively.Western-blot analysis showed that the four McAbs could react with the GPV recombinant VP3 protein specifically.IFA showed that the four McAbs could combine with the natural GPV.