Shrimp White Spot Syndrome Virus (wssv) Monoclonal Antibodies And Characteristics

White Spot Syndrome Virus (WSSV) is the most harmful and the most influencing shrimp virus, but no medicament and vaccine against WSSV are produced up to the present. The neutralizing monoclonal antibody was reported close relative with the protective antigen of WSSV, this study planed to produce monoclonal antibodies against WSSV and selected the neutralizing monoclonal antibodies against WSSV which will accelerate the advancement of basal theory of WSSV and offer theory foundation and practice guidance to develop idiotypic vaccine, synthetic peptide vaccine and genetically engineered vaccine.Neat and high density of WSSV could be purified and gained by use of sucrose grads' centrifuge, and the viral structure of WSSV was analysed by electron microscope and SDS-PAGE gel electrophoresis. 4 weeks Balb/c mouse was immunized with WSSV of shrimp; polyethylene glycol (PEG) was used as solution for cell fusion of WSSV-immune spleen cells with P3-X63-Ag8Ul myeloma cells, and hybridomas were selected by means of Immunofluorescence assays (IFA). 22 strains of hybridomas producing positive antibodies against WSSV of shrimp were gained and eight of them were cloned .A total of 16 monoclonal antibodies, 8 were produced in this experiment and the other 8 monoclonal antibodies were produced previously in the laboratory, were used in this trial of selecting neutralizing monoclonal antibodies. Crayfish (Cambarus proclarkii) as a host instead of penaeid shrimp was infected with WSSV to research monoclonal antibody neutralizing activity in vivo. Meantime, the neutralizing activity of the mixture of monoclonal antibodies and the effect of mixing to the neutralizing activity were studied. The neutralizing trial of monoclonal antibodies in vivo was proved by observing the mortality after injected with the mixture of monoclonal antibody and WSSV solution, and by detecting WSSV with the method of immunofluorescence antibody technique. The results of the neutralizing trial in vivo showed obviously: every monoclonal antibody possessed different intension neutralizing activity against WSSV; the neutralizing activities of the monoclonal antibodies 2B2, 2D2 and 4F3 were the best among the 16 monoclonal antibodies; the neutralizing activity declined obviously after mixing the monoclonal antibodies.