细胞工厂在乙型脑炎减毒活疫苗生产中的应用

Application of cell factory in production of live attenuated Japanese encephalitis vaccine

目的采用40层细胞工厂(CF40)替代15 L转瓶工艺制备乙型脑炎(乙脑)减毒活疫苗。方法分别采用CF40和15 L转瓶工艺制备乙脑减毒活疫苗.通过显微镜观察和细胞计数来考察细胞质量。病毒收获时观察细胞病变情况,并对2种工艺制备的原液、半成品以及成品进行病毒滴度检测对比,成品37℃放置7d考察成品热稳定性。结果两种工艺相比,CF40细胞密度高于15L转瓶,差异有统计学意义(F=10.60,P<0.05),生长一致性和单只地鼠制备的细胞数量也均优于15L转瓶;CF40和15 L转瓶制备的单一病毒收获液平均滴度分别为7.59和7.56 lg噬斑形成单位/ml,差异无统计学意义(F=0.63,P>0.05),但CF40单一病毒收获液收率一致性更好;两种工艺制备的原液、半成品、成品滴度以及7 d后37℃热稳定性均符合企业注册标准规定。结论使用CF40制备乙脑减毒活疫苗是可行的,并且在产品收率和一致性方面优于15 L转瓶工艺。

Objective To prepare live attenuated Japanese encephalitis vaccine(JEV)by 40-layer cell factory(CF40)instead of 15 L rolling bottle.Methods JEV was prepared by CF40 and 15 L rolling bottle separately.Cell quality was checked by microscope and cell counting.Growth of cells was observed when virus was harvested.Virus titers of the bulk,final bulk and final product prepared by two processes were compared.The thermal stability of the final product was investigated after being placed at 37℃for 7 days.Results Between two processes,the cell density in CF40 was significantly higher than that in 15 L rolling bottle(F=10.60,P<0.05).The growth consistency and the number of cells prepared per hamster were better in CF40.The average virus titers of single harvest solutions from CF40 and 15 L rolling bottle were 7.59 and 7.56 lg plaque-forming unit/ml,respectively,with no significant difference(F=0.63,P>0.05),but the yield of CF40 single virus harvested solution was more consistent.The titers of the bulk,final bulk and final product prepared by the two processes,and the thermal stability at 37℃after 7 days of final product all met the provisions of enterprise registration standards.Conclusion It is feasible to prepare live attenuated JEV by CF40,and its yield and consistency of product are better than those of 15 L rolling bottle.