腮腺炎病毒在转瓶和细胞工厂中制备的比较

Comparison of live attenuated mumps vaccine prepared in rotating bottle and cell factory

目的比较15 L转瓶及40层细胞工厂工艺制备的腮腺炎减毒活疫苗致细胞病变情况、病毒滴度和原液各项质量指标.方法分别采用15 L转瓶及40层细胞工厂培养原代鸡胚细胞1~3 d后感染S79株腮腺炎病毒,继续培养5~7 d收获病毒液.两种培养方式收获的腮腺炎病毒单次收获液分别合并后获得疫苗原液,并进行相关检定.结果转瓶收获的单次病毒液0和21 d检定的平均滴度分别为6.6、6.1 lg半数细胞培养感染量(50%cell culture infective dose,CCID50)/ml.细胞工厂收获的单次病毒液0和21 d检定的平均滴度分别为6.8、6.2 lgCCID50/ml.两种方式制备的腮腺炎减毒活疫苗原液各项质量指标均合格.结论应用40层细胞工厂工艺制备的腮腺炎减毒活疫苗致细胞病变情况及单次病毒收获液滴度均优于15 L转瓶培养工艺,此实验为40层细胞工厂制备腮腺炎减毒活疫苗的大规模生产及工艺改进奠定了基础.

Objective To compare the cytopathic conditions, virus titers and quality indicators of live attenuated mumps vaccine prepared in 15 L rotating bottle and 40-layer cell factory. Methods Primary chicken embryo cells were cultured in 15 L rotating bottle and 40-layer cell factory, respectively. Cells were infected by the S79 mumps virus strain after 1-3 d of culture. And the viruses were harvested after 5-7 d further culture. The single harvest virus liquids harvested by 2 methods were combined seperatedly to be the vaccine bulks. These vaccine bulks were subjected to control tests. Results The mean titer of virus cultured in rotating bottle was 6.6 lg 50% cell culture infective dose (CCID50) per ml on 0 d, and 6.1 lgCCID50 per ml on 21 d. The mean titer of virus cultured in cell factory was 6.8 lgCCID50/ml on 0 d, and 6.2 lgCCID50/ml on 21 d. All the quality indexes of live attenuated mumps vaccine prepared by 2 methods met the relevant requirements. Conclusions The cytopathic conditions and virus titers of live attenuated mumps vaccine prepared in cell factory are better than those in rotating bottle. The study lays foundation for large-scale production and technological improvement of live attenuated mumps vaccine prepared in cell factory.