抗WSSV囊膜蛋白(VP28和VP19)单抗的体内中和效果研究

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

通过酶联免疫吸附法(ELISA)测定不同稀释度对虾白斑综合征病毒(WSSV)与已制备的WSSV囊膜蛋白单克隆抗体结合的OD值。利用克氏原螯虾Cambarus proclarkii动物模型,将不同稀释度病毒与单抗1∶1混合孵育2h后,肌肉注射克氏原螯虾(50μl/只),观察记录螯虾的死亡情况。ELISA结果显示,在1×10-3病毒稀释度下两种单抗均足量。在螯虾体内中和实验中,当病毒浓度为1×10-3、1×10-4、1×10-5和1×10-6稀释度时,MAb1D6(VP28)螯虾组最终死亡率分别为100%、90%、16.7%和6.7%,而MAb2E9(VP19)螯虾组最终死亡率分别为100%、100%、100%和93.3%。这表明随病毒浓度的降低,MAb1D6(VP28)的中和效果越明显。而MAb2E9(VP19)并无明显的中和效果。

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^-5and 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.