大规模区带离心纯化Vero细胞乙脑疫苗  

Large Scale Purification of Inactivated Japanese Encephalitis Vaccine from Vero Cells by Zonal Centrifugation

作　　者：石慧颖[1] 丁志芬[1] 赵敏[1] 庞成华[1] 杨抗抗[1] 李荆[1] 

机构地区：[1]卫生部北京生物制品研究所

出　　处：《中国病毒学》1998年第3期208-213,共6页Virologica Sinica

基　　金：国家"八五"科技攻关项目

摘　　要：本文报道一种适合疫苗生产的大规模纯化Ｖｅｒｏ细胞乙脑疫苗的方法。原疫苗经适当浓缩和去除ＤＮＡ处理后，用不连续蔗糖梯度（３６％和６０％）。３２６００ｇ，速率区带离心４ｈ。纯化后疫苗的效力比中国参考疫苗高出６倍以上，补体结合抗原比中国参考疫苗高４～８倍。总蛋白含量低于３０μｇ／ｍＬ，牛血清含量降至０．５μｇ／ｍＬ以下，细胞残余ＤＮＡ低于１００ｐｇ／０．５ｍＬ。用此法连续制备三批纯化疫苗，其纯度和效力均高于日本鼠脑纯化疫苗。此法对于制备其它纯化Ｖｅｒｏ细胞疫苗也具有一定的参考意义。Japanese Encephalitis (JE) Vaccine in Vero cell can be easily purified by zonal centrifugation at non continuous sucrose gradients (36% and 60%), 32 600g for 4h. The calf serum protein and other nonviral proteins in the vaccine were almost separated from the JE virus. The residual calf serum protein was less than 0.5μg/mL, and the total protein was less than 30μg/mL. The residual Vero cell DNA in the vaccine was less than 100 pg/0.5mL. The titer of purified Japanese Encephalitis vaccine is six times higher than China control vaccine. This method is recommended as an available method to purify JE vaccine from Vero cell in large-scale because it is simple, rapid and inexpensive.

关 键 词：乙脑疫苗 区带离心 VERO细胞 纯化乙脑疫苗 

分 类 号：R512.320.3[医药卫生—内科学]