Preparation of Rabies Vaccine for Human Use with Vero Cells in Basket Bioreactor

Objective To prepare rabies vaccine for human use with Vero cells in basket bioreactor.Methods Rabies vaccine virus strain aGV was cultured in Vero cells in 14 L bioreactor(500 g of disk carriers in basket) by perfused culture.The continuously harvested virus liquid was concentrated by ultrafiltration,inactivated,and purified by column chromatography,then added with stabilizer and prepared into freeze-dried vaccine.The prepared vaccine was determined for titers before and after lyophilization and after storage at 37℃ for 28 d.Results The cell density reached 1.0 × 107 cells / ml at most,and the titer of harvested virus liquid was 7.5 × 106 ～ 3.4 × 108 FFU / ml.After purification,the total protein content,residual Vero cell protein content,residual Vero cell DNA content and vaccine titer were less than 80 μg / 0.5 ml,less than 3.5 μg / 0.5 ml,less than 100 pg / 0.5 ml and more than 5.0 IU / 0.5 ml.Conclusion Basket bioreactor may be used for preparation of rabies vaccine for human use with high quality.