| Literature DB >> 19324009 |
Fang-Feng Chiu1, Nandini Venkatesan, Chia-Rong Wu, Ai-Hsiang Chou, Hsin-Wei Chen, Shu-Pei Lian, Shih-Jen Liu, Chin-Cheng Huang, Wei-Cheng Lian, Pele Chong, Chih-Hsiang Leng.
Abstract
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 6M 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.Entities:
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Year: 2009 PMID： 19324009 DOI： 10.1016/j.bbrc.2009.03.106
Source DB: PubMed Journal: Biochem Biophys Res Commun ISSN： 0006-291X Impact factor: 3.575