冻干人用狂犬病疫苗(地鼠肾细胞)生产毒种的筛选  

Screening of virus seeds for production of freeze-dried rabies vaccine(primary hamster kidney cells) for human use

作　　者：任红卫[1] 郑学刚[1] 李守丽[1] 金学敏[1] 杨淼[1] 李国晏[1] 何彦林[1] 

机构地区：[1]兰州兰生血液制品有限公司兰州生物制品研究所有限责任公司甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《中国生物制品学杂志》2017年第7期702-705,共4页Chinese Journal of Biologicals

摘　　要：目的筛选冻干人用狂犬病疫苗(地鼠肾细胞)生产毒种。方法将原代地鼠肾细胞传毒种(北京固定毒7aG)和豚鼠脑传毒种(北京固定毒6aG4)平行接种于10层细胞工厂培养的原代单层地鼠肾细胞,收获病毒液,共收获8次,1、3、5、7、9号细胞工厂接种脑毒,2、4、6、8、10号细胞工厂接种细胞毒。1次收获(简称1收)培养72h,从2次收获(简称2收)开始,培养24h,收获液经超滤浓缩、柱层析纯化、除菌过滤、再浓缩配制成人用狂犬病疫苗半成品,检测各项指标,筛选最佳毒种。结果6aG4株毒种制备的病毒收获液、浓缩液、纯化液、纯化除菌过滤液、狂犬病疫苗半成品的各项检定指标都优于7aG毒种。结论豚鼠脑传毒种(6aG4)明显优于原代地鼠肾细胞传毒种(7aG),选择豚鼠脑传毒种(6aG4)作为人用狂犬病疫苗(地鼠肾细胞)生产毒种。Objective To screen the virus seeds for production of freeze-dried rabies vaccine（primary hamster kidney cells）. Methods Primary hamster kidney cells cultured in 10 layers of cell factory were inoculated with Beijing fixed virus 7aG subcultured in primary hamster kidney cells and Beijing fixed virus 6aG4 subcultured in guinea pig brain cells respectively. The virus liquids were harvested for 8 times, of which those subcultured in guinea pig brain cells were inoculated into cell factories of No. 1, 3, 5, 7 and 9, while those subcultured in primary hamster kidney cells into cell factories of No. 2, 4, 6, 8 and 10. The first virus harvest was obtained 72 h after culture, while the second to the eighth virus harvests were obtained each at an interval of 24 h. The harvested liquids were concentrated by ultrafiltration, purified by column chromatography, sterilized by filtration, and further concentrated to prepare the final bulk of rabies vaccine for adult use, which was subjected to control tests for screening of optimal virus seeds. Results All the indexes of harvested liquid, concentrated liquid, purified liquid, filtrate and final bulk of rabies vaccine prepared with strain 6aG4 were superior to those of 7aG strain. Conclusion The virus seeds subcultured in guinea pig brain cells was superior to that in primary hamster kidney cells for production of rabies vaccine（primary hamster kidney cells）for human use.

关 键 词：冻干人用狂犬病疫苗(地鼠肾细胞) 原代地鼠肾细胞传毒种(7aG) 豚鼠脑传毒种(6aG4) 超滤浓缩 柱层析纯化 

分 类 号：R373.9[医药卫生—病原生物学] R392-33[医药卫生—基础医学]