Preparation and preliminary application of a neutralizing monoclonal antibody against rabies virus

Abstract

Objective To prepare a neutralizing monoclonal antibody against rabies virus. Methods BALB/c mice were immunized with the inactivated rabies virus CTN strains on day 0, 7, 14 and 28. Spleen cell samples were collected and then fused with SP2/0 cells to prepare the hybridoma cell line. Positive hybridoma cells that were screened out with RFFIT technique were injected into BALB/c mice intraperitoneally. Ascites samples were collected from the mice to separate neutralizing monoclonal antibodies. Affinity chromatography was used for the purification of neutralizing monoclonal antibodies. Subtype identification and sequencing analysis were performed for further identification. A colloidal gold strip based method for rapid detection of rabies vaccine was established with the prepared monoclonal antibodies. Results The hybridoma cell line, CTN-McAb1, was prepared successfully with stable secretion of neutralizing monoclonal antibodies against rabies virus. The purity of those antibodies was more than 95% after purification and the subtype of them was IgG1. The colloidal gold strip for raid detection of rabies vaccine was successful prepared. Conclusion The neutralizing monoclonal antibody against rabies virus was successfully prepared and could be used for preliminary application. This study will be of great significance for the quality control of rabies vaccine. Key words: Rabies virus; Monoclonal antibody; Neutralizing activity