Separation，Purification and Identification of Structural Proteins of Hepatitis A Virus

Lei-Yin-Beng; LIN Yu-Lin; TAO Hua-Jun; JIAN Ai-Zhen; LI Hui; ZHANG Wei-Yang

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June 1999

Lei-Yin-Beng, LIN Yu-Lin, TAO Hua-Jun, JIAN Ai-Zhen, LI Hui and ZHANG Wei-Yang. Separation，Purification and Identification of Structural Proteins of Hepatitis A Virus[J]. Virologica Sinica, 1999, 14(3): 210-216.

Citation: Lei-Yin-Beng, LIN Yu-Lin, TAO Hua-Jun, JIAN Ai-Zhen, LI Hui, ZHANG Wei-Yang. Separation，Purification and Identification of Structural Proteins of Hepatitis A Virus .VIROLOGICA SINICA, 1999, 14(3) : 210-216.

甲型肝炎病毒结构蛋白的分离纯化及鉴定

1.
同济医科大学附属协和医院病毒室,武汉430022
2.
湖北医科大学病毒研究所，武汉430071

摘要

利用高效持续带毒增殖的细胞株传代增殖获取甲型肝炎病毒（ＨＡＶ）。采用不连续蔗糖／甘油密度梯度超速离心分离高度纯化的ＨＡＶ颗粒。温和病毒裂解液裂解病毒后，运用高效液相色谱分离出三种蛋白，ＳＤＳＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ证实所分离蛋白为ＨＡＶ三种主要结构蛋白ＶＰ１、ＶＰ２和ＶＰ３，蛋白含量较高。
- 甲型肝炎病毒
- , 结构蛋白
- , 分离
- , 纯化
- , 鉴定

Separation，Purification and Identification of Structural Proteins of Hepatitis A Virus

1.
Department Virology，Union Hospital Tongfi Medical University，Wuhan 430022
2.
Virtu Rts~rch Instituy, Hubel Medical University, Wuhann 430071

Abstract

A cell line persistently infected by Hepatitis A virus (HAV) was ultilized to produce high yield viral particles of HAV. HAV particles were recovered in a single ultracentrifugation step through discontinuous sucrose/glycerol density gradients. Three kinds of major structural protein of HAV were separated and purified by high performance liquid chormatography. SDS PAGE demonstrated the molecular weight of three structural proteins of HAV being 33, 29 and 27?kD. The special immune reactions were shown in Western blot test, suggesting the proteins with 33, 29 and 27?kD are VP\-1, VP\-2 and VP\-3 of the virus, respectively. The total contents of the proteins, VP\-1, VP\-2 and VP\-3 were respectively 687, 220.8, 68 and 213.6?g from 11?200?cm\+2 cells infected with HAV. The highly purified particles and sturctural proteins of HAV may be used in research for the development of vaccine, clinical diagnosis and pathogenesis of HAV.
- Hepatitis A virus
- , Structural proteins
- , Separation
- , Purifica tmn
- , Identification

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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

Separation，Purification and Identification of Structural Proteins of Hepatitis A Virus

1. Department Virology，Union Hospital Tongfi Medical University，Wuhan 430022
2. Virtu Rts~rch Instituy, Hubel Medical University, Wuhann 430071

Abstract: A cell line persistently infected by Hepatitis A virus (HAV) was ultilized to produce high yield viral particles of HAV. HAV particles were recovered in a single ultracentrifugation step through discontinuous sucrose/glycerol density gradients. Three kinds of major structural protein of HAV were separated and purified by high performance liquid chormatography. SDS PAGE demonstrated the molecular weight of three structural proteins of HAV being 33, 29 and 27?kD. The special immune reactions were shown in Western blot test, suggesting the proteins with 33, 29 and 27?kD are VP\-1, VP\-2 and VP\-3 of the virus, respectively. The total contents of the proteins, VP\-1, VP\-2 and VP\-3 were respectively 687, 220.8, 68 and 213.6?g from 11?200?cm\+2 cells infected with HAV. The highly purified particles and sturctural proteins of HAV may be used in research for the development of vaccine, clinical diagnosis and pathogenesis of HAV.