Large-scale purification of inactivated Japanese encephalitis vaccine from Veto cells by zonal centrifugation

DAN Hui-Ying; DING Zhi-Fen; DIAO Min; LONG Cheng-Hua; YANG Kang-Kang; LI Jing

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June 1998

DAN Hui-Ying, DING Zhi-Fen, DIAO Min, LONG Cheng-Hua, YANG Kang-Kang and LI Jing. Large-scale purification of inactivated Japanese encephalitis vaccine from Veto cells by zonal centrifugation[J]. Virologica Sinica, 1998, 13(3).

Citation: DAN Hui-Ying, DING Zhi-Fen, DIAO Min, LONG Cheng-Hua, YANG Kang-Kang, LI Jing. Large-scale purification of inactivated Japanese encephalitis vaccine from Veto cells by zonal centrifugation .VIROLOGICA SINICA, 1998, 13(3) : 213.

大规模区带离心纯化Vero细胞乙脑疫苗

1.
卫生部北京生物制品研究所

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３０ｇ／ｍＬ，牛血清含量降至０．５ｇ／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。
- 区带离心 Vero细胞纯化乙脑疫苗

Large-scale purification of inactivated Japanese encephalitis vaccine from Veto cells by zonal centrifugation

1.
National Vaccine and Serum Institute，Beijing 100024

Abstract

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 0.5g/mL, and the total protein was less than 30 g/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.
- Zonal centrifugation Vero cell Purified Japanese Encephalities vaccine

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Large-scale purification of inactivated Japanese encephalitis vaccine from Veto cells by zonal centrifugation

1. National Vaccine and Serum Institute，Beijing 100024

Abstract: 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 0.5g/mL, and the total protein was less than 30 g/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.