In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish(Cambarus proclarkii)

The neutralizing activities of eight monoclonal antibodies (MAbs) against white spot syndrome virus (WSSV) (2D2, 2B2, 1D2, 1D5, 1C2, 4A1, 6A4 and 6B4) were analyzed by in vivo experiments.Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS, and then incubated with MAbs (hybridoma culture supernatant), respectively.The mixture of WSSV and MAbs were injected into crayfish (Procambarus clarkii).After challenge, the death rates of crayfish were counted to determine the neutralizing activities of MAbs.At the same time, the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control, respectively.The results showed that, at each virus dilution, the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control, though they all showed 100% mortality within 25 d, and meanwhile, few crayfish died in the negative control.Among the eight MAbs, 2D2, 2B2, 1D2 and 1D5, especially the former two, delayed the mortality significantly, and 1C2, 4A1 and 6A4 delayed the mortality as well but not so efficiently, while MAb 6B4 was efficient only when the virus concentration increased.The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.