Immunological study of HA1 domain of hemagglutinin of influenza H5N1 virus

被引：47

作者：

Chiu, Fang-Feng ^{[1
]}

Venkatesan, Nandini ^{[1
]}

wu, Chia-Rong ^{[1
]}

Chou, Ai-Hsiang ^{[1
]}

Chen, Hsin-Wei ^{[1
]}

Lian, Shu-Pei ^{[1
]}

Liu, Shih-Jen ^{[1
]}

Huang, Chin-cheng ^{[3
]}

Lian, Wei-Cheng ^{[1
,2
]}

Chong, Pele ^{[1
]}

Leng, Chih-Hsiang ^{[1
]}

机构：

[1] Natl Hlth Res Inst, Vaccine Res & Dev Ctr, Zhunan 350, Miaoli, Taiwan

[2] Ctr Dis Control, Taipei 105, Taiwan

[3] Council Agr, Anim Hlth Res Inst, Taipei 251, Taiwan

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2009年 / 383卷 / 01期

关键词：

Influenza hemagglutinin; Oxidative folding; H5N1 neutralizing antibody; ESCHERICHIA-COLI; A VIRUSES; EXPRESSION; BINDING; PROTEIN; CONSTRUCTION; PURIFICATION; SUBTYPES; FUSION; SITES;

D O I：

10.1016/j.bbrc.2009.03.106

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The neutralization titer of a hemagglutinin (HA)-specific neutralizing antibody against new isolates reflect both the antigenic drift and the conformation status of HA protein in these new influenza viruses. Since most antigenic sites are in the HA1 domain of HA, using HA1 domain of influenza virus as antigen is of great importance in vaccine development. In this study, we investigate different Purification processes for optimizing the immunological properties of an Escherichia coli-expressed HA1 domain (rH5HA1) of influenza H5N1 virus. rH5HA1 was expressed as inclusion bodies and extracted with 6 M guanidine hydrochloride (GnHCl)/PBS buffer. The best condition for generating HA1-specific neutralization determinants is on-column oxidative refolding procedures with GSH/GSSG and L-arginine buffer. Others refolding procedures Such as using high-pH buffer and/or different detergent solubilizations were found to be ineffective producing neutralization epitope recognized by a HA1-specific neutralizing monoclonal antibody that was raised against H5N1 virus. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：27 / 31

页数：5

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