Preparation and preliminary application of monoclonal antibodies against VP2 COE protein of infectious pancreatic necrosis virus

The recombinant protein IPNV VP2 was used as immunogen after purification by Ni-NTA.The 8-week-old BALB/c mice were intraperitoneally immunized with the VP2 protein for three times,then myeloma cells SP2/0 were fused with the spleen cells of the immunized BALB/c mice.Two hybridoma cell lines against the VP2 protein were obtained by screening with the indirect ELISA and limiting dilution assay,which were identified to be IgG1 subtype and named 5G10,5F3.The numbers of chromosomes of the two hybridomas were in the range of 75 to 120.Their antibody titers of cells culture supernatant were 1∶ 105,1∶ 102 respectively.Their titers of ascites were 1∶ 108,1∶ 104 respectively.Western-blot analysis and indirect immunofluorescence showed that the two McAbs could react with IPNV specificity.The 2 McAbs had no reactive capability with IHNV,VHSV,SVCV and HRV by the indirect ELISA.Antibodies additivity assay demonstrated that 5G10 and 5F3 recognized the different epitopes of IPNV VP2 nucleoprotein.We detected the clinical suffering from IPN rainbow trout liver tissue material,and the results confirmed that the 2 McAbs can be used for follow-up testing.