Preparation of mouse anti-human rotavirus VP7 monoclonal antibody and its protective effect on rotavirus infection

摘要

The aim of the current study was to prepare and identify mouse anti-human rotavirus (RV) VP7 monoclonal antibodies and explore their protective effects on RV infection. The mouse anti-human RV VP7 monoclonal antibody was produced using the ascites method and identified via western blot analysis. In vitro neutralization of mouse anti-human RV VP7 monoclonal antibodies was detected by performing an MTT assay. The TCID50 value was calculated to obtain antibody neutralization titers. A mouse RV infection model was generated to assess the protective effect of the mouse anti-human RV VP7 monoclonal antibody in experimental animals. Monoclonal antibodies were successfully prepared and their purity reached ≥90%. Western blotting demonstrated that monoclonal antibodies specifically bound to the purified Wa RV strain, with a specific reaction band at ~40 kDa. Monoclonal antibody in vitro neutralization results demonstrated that cell survival rate in the virus + monoclonal antibody group was higher than that in virus + maintenance fluid group (P<0.05). Monoclonal antibody neutralization titer detection revealed that the cytopathic effect did not extend beyond 4 days. In addition, the calculated monoclonal antibody neutralization titer was 1:446. The results revealed that the positive rate of colloidal gold RV in the 100 µl monoclonal antibody group was significantly lower than that in the control group (P<0.05). Furthermore, the protection rate of the 100 µl monoclonal antibody group was 71.4%, whereas the 50 µl monoclonal antibody group was 42.9% and the ribavirin group was 57.1%. In conclusion, the results of the current study demonstrated that mouse anti-human RV VP7 monoclonal antibodies can be successfully prepared using ascites method. These antibodies also effectively neutralize the cytotoxic effects of the human RV Wa strain in vitro and mouse anti-human RV VP7 monoclonal antibodies also exhibited a good protective role in mice. Furthermore, greater protective effects were observed at a higher dose and the protective effects of these high dose treatments were superior to that of ribavirin.