Purification and Identification of Inactivated Enterovirus 71 Vaccine Derived from Vero Cells

OBJECTIVE To develop a purification procedure of inactivated enterovirus 71 (EV71) vaccine derived from Vero cells. METHODS The harvested liquid was pretreated by means of centrifugation, ultrafiltration, inactivated with formmdehyde, and then the inactivated EV71 vaccine was purified by Sephacryl S-400 HR gel filtration chromatography and Source 30Q ion exchange chromatography. The vaccine was subjected to control tests and analysis then sterilized by filtration. RESULTS A1l test results of three lots vaccine after purification accorded with the requirements of Ch.P(2010). All the removal rates of protein in vaccine were >99.9%. Typical morphology of EV71 was observed in purified EV71 samples under transmission electron microscope．SDS-PAGE analysis showed three protein straps with relative molecular weight which was according with the VP1, VP2 and VP3 of EV71 and could be confirmed by Western blot analysis. The purified vaccine induced neutralizing antibody titer in rats. CONCLUSION A purification procedure of EV71 vaccine derived from Vero cells is developed, and the vaccine demonstrates high immunogenicity.