In vivo neutralization assay of monoclonal antibodies specific to WSSV envelope protein (VP28 and VP19)

The optical density (OD) was measured by Enzyme-Linked Immunosorbent Assay (ELISA) with the combination of different dilutions of white spot syndrome virus (WSSV) and monoclonal antibodies (MABS) specific to WSSV envelope protein. As the model animal,crayfishes Cambarus proclarkii were intramuscular injected of 50μl WSSV at different dilutions pre-incubated with MABS,then their cumulative mortalities were recorded. Results of ELISA showed that both MAb1D6 (VP28) and MAb2E9 (VP19) were sufficient at 1×10-3 virus dilution. In vivo neutralization assay showed mortalities in the MAb1D6 (VP28) group were 100%,90%,16.7% and 6.7%,at a dilution of 1×10-3,1×10-4,1×10-5 and 1×10-6,respectively;while mortalities in the MAb2E9 (VP19) group were 100%,100%,100% and 93.3%,respectively. These results demonstrated that the neutralization effect of MAb1D6 was concentration dependent. MAb2E9 has no obvious neutralization effect.