Preparation And Detection Of Monoclonal Antibodies To Channel Catfish Virus And Polyclonal Antibodies To Serum Immunoglobulin

Channel catfish virus disease outbreaks can cause high mortality of channel catfish major economic losses. Currently, there is no effective drug for the disease, detect and clear pathogens is the only way to treat the disease, which requires a rapid detection method.Channel catfish virus and serum were studied in the paper. The mouse anti-virus monoclonal antibody and rabbit anti-serum polyclonal antibody were prepared for the Immunology.A large number of viruses were obtained by cell proliferation, through ultracentrifugation, and the purified virus concentration was 4mg/ml. Using purified virus as the immunogen, in accordance with the immunization program, six monoclonal antibodies against the virus were prepared, named 1F1,1B7,3E4,3G10, 4C4 and 4D4. The titer of three monoclonal antibodies can be achieved 1:256000, respectively,1F1,1B7, and 3E4, and the other three slightly lower potency. Immunofluorescence experiment showed that monoclonal antibodies prepared with good specificity. In addition to 3E4, the five blocking monoclonal antibodies can stop virus infecting cells. By immunoblotting experiments, the molecular weight of protein which were bind by monoclonal antibodies was detect, and the according to the published viral gene sequences and predicted protein mapping, the name was obtained too, which provided the basis for further research, there are ORF1, ORF43, ORF56, ORF57.Taking the channel catfish serum as immunogen, which was purified by ammonium sulfate precipitation and column chromatography, rabbit anti-serum polyclonal antibody was got, and the ELISA titer can be achieved to 1:64000.The detection of the antibody lays a foundation for the virus.