| All around of the world, respiratory disease infected by viral has become the common disease. Human Bocavirus is one of classical perpetrators of respiratory infection, it was found later, but was paid attention by scientific research workers. The efficient title of antibody is useful for the effective diagnosis and treatment of antigen, to establish the system method, monoclonal antibody is a main tool of detection and diagnosis of the pathogen. There was few reports of the study about HBoV monoclonal antibody, so producing monoclonal antibody has significance for the diagnosed and therapy of the antigen.The recombinant prokaryotic expression vector pET-30a was constructed in this study, and the recombinant VP2protein was expressed by E. coli, and the protein was purified and denaturated using the column. Then, the antigen in serum was detedted The BALB/c mouse was immunized using purified protein as antigen. The positive hybridoma cells were selected with hybridoma technique and ELISA assay. The monoclonal antibody was induced in vivo and identified by isotype and titerThe recombinant plasmid was obtained, and the purified HBoV VP2protein was produced. The antigen was detected using respiratory tract infection patients serum antigen detection, positive rate was77.7%. According to the domestic and international reports, the detection result showed that VP2protein is a good antigen. The mouse produced ascites of monoclonal antibody, induced by the cell line. According to the detection, the monoclonal antibody was identified as IgG, and the obtained antibody has a better ability to combine antigen, and the titer of the monoclonal antibody was up to1:4×105.Monoclonal antibody was obtained in this experiment, and it is helpful for the further study of human bocavirus, and monoclonal antibodies are the main tool for pathogen detection and diagnostics. The experimental results laid the foundation for human bocavirus diagnosis and treatment. |
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