[Preparation and characterization of monoclonal antibodies against viral hemorrhagic septicemia virus].

OBJECTIVE

To prepare the monoclonal antibody (mAb) against viral hemorrhagic septicemia virus (VHSV) and analyze the biological properties.

METHODS

The BALB/c mice were immunized with the VHSV which was purified by differential centrifugation. Spleen cells of the immune mice were collected and fused with Sp2/0 myeloma cells by hybridoma technology. The indirect ELISA was used to screen hyridoma cells and identify the specificity of mAb. The Western blotting was used to identify antigen site.

RESULTS

After three cycle of subcloning with limited dilution method, we obtained a cell strain that secreted mAb against VHSV and was named 4A5. The indirect ELISA showed that the titer of mAb ascites was 10⁻⁴;. It belonged to IgG3 subclass κ chain and couldn't react with other fish viruses or fish cell lines in the ELISA except VHSV. Western blotting revealed that mAb 4A5 bound to a 70 kDa of protein band of VHSV as antigen.

CONCLUSION

The obtained mAb has a high specificity, which could be used for rapid diagnosis and detection of VHSV in aquaculture.