White Spot Syndrome Virus Monoclonal Antibody Production And Application

The white spot syndrome virus (WSSV) is the most dangerous virus to shrimp aquaculture worldwide in recent years. Because of lacking the susceptible cell lines, the mechanism of virus infection, replication and assembly remains unknown, and this in turn hamper the effective control to this viral disease. VP28 is the most abundant envelope protein and was found to play an important role in WSSV infection. In this thesis, the purified WSSV particles were used to immunize the BALB/c mice, and the expressed VP28 and purified WSSV were used to screen the monoclonal antibody (Mab) by ELISA. Six Mabs against VP28 and one Mab against an unknown protein of WSSV were identified. The western blot and dot blot showed that all the six Mabs is against conformational epitope of VP28. A sandwich ELISA was developed using Mabs 3A10 and 1E7 which could detect at least 8.5×106 purified virus, and 17.8ng purified VP28. An in vitro neutralization experiment to WSSV with the 6 Mabs showed that 4 of them could delay the initial mortality of crayfish infected by WSSV, suggesting that these Mabs can be used for further function study of VP28.The Mab 7B4 was labeled with colloidal gold particles and used to localize the VP28 on virus envelope by immunogold labelling. Besides, three WSSV protein VP36A,VP36B and VP31 which contain the cell attachment motif were confirmed to localize on virions by immunogold labeling.