title: Immunological study of HA1 domain of hemagglutinin of influenza H5N1 virus.
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.
authors.affiliation: Vaccine Research and Development Center, National Health Research Institutes, Zhunan, Miaoli, Taiwan, ROC.
authors.forename: Fang-Feng
authors.initials: FF
authors.lastname: Chiu
authors.forename: Nandini
authors.initials: N
authors.lastname: Venkatesan
authors.forename: Chia-Rong
authors.initials: CR
authors.lastname: Wu
authors.forename: Ai-Hsiang
authors.initials: AH
authors.lastname: Chou
authors.forename: Hsin-Wei
authors.initials: HW
authors.lastname: Chen
authors.forename: Shu-Pei
authors.initials: SP
authors.lastname: Lian
authors.forename: Shih-Jen
authors.initials: SJ
authors.lastname: Liu
authors.forename: Chin-Cheng
authors.initials: CC
authors.lastname: Huang
authors.forename: Wei-Cheng
authors.initials: WC
authors.lastname: Lian
authors.forename: Pele
authors.initials: P
authors.lastname: Chong
authors.forename: Chih-Hsiang
authors.initials: CH
authors.lastname: Leng