Preparation And Primary Application Of Monoclonal Antibody Against Rabies Virus

Rabies is an age-old infectious disease of the central nervous system of major importance to human and veterinary medicine. It causes a fatal encephalomyelitis for which there is no treatment once the clinical symptoms have appeared. Vaccination is the only way to combat the infection. Nowadays the pathogenic mechanisms have been the subject of many studies, but are still not well understood. We need to know the pathogenic gene in order to root out the rabies.In this study, Rabies virus cell adapted strain ERA, grown in monolayer cultures of BHK-21 cells and purified by centrifugation , Zn(AC)2 precipitation and subsequently 10~55%(w/v) sucrose density grasient . The purification have rabies virus of highly titer by RT-PCR. BALB/c mouse were immunized with the purified RV, mouse spleen cell were fused with myelomas SP2/0. Culture supernatants of hybridomas were screened by indirect ELISA and immobilization ELISA, positive colonies were cloned three times by limited dilution, five cell line while secrete McAbs against rabies virus were obtained. There five McAbs designated as E1-1, E1-2, E1-4, E1-5 and E1-6 .The hybridomas were inoculated in the mice bellies to produce McAbsand the ascites McAbs titer had been measured between 1:2 000 to 1:8 000.All McAbs could react with four rabies virus strains(119,BM,LA,F-)collected from different county in Guangxi, indicating that five McAbs are anti-rabies virus. The McAbs did not cross react with PRRSV, HCV, PCVand PRV, demonstrated good specificity. Three of five McAbs (E1-1 , E1-2 , E1-5)have highly cellular neutralizing abilities .