Immunological study of HA1 domain of hemagglutinin of influenza H5N1 virus

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...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2009-05, Vol.383 (1), p.27-31
Main Authors: Chiu, Fang-Feng, Venkatesan, Nandini, Wu, Chia-Rong, Chou, Ai-Hsiang, Chen, Hsin-Wei, Lian, Shu-Pei, Liu, Shih-Jen, Huang, Chin-Cheng, Lian, Wei-Cheng, Chong, Pele, Leng, Chih-Hsiang
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Antibodies, Monoclonal - immunology
Cloning, Molecular
Escherichia
Escherichia coli - genetics
H5N1 neutralizing antibody
Hemagglutinin Glycoproteins, Influenza Virus - chemistry
Hemagglutinin Glycoproteins, Influenza Virus - genetics
Hemagglutinin Glycoproteins, Influenza Virus - immunology
Humans
Influenza A Virus, H5N1 Subtype - chemistry
Influenza A Virus, H5N1 Subtype - genetics
Influenza A Virus, H5N1 Subtype - immunology
Influenza hemagglutinin
Influenza Vaccines - chemistry
Influenza Vaccines - genetics
Influenza Vaccines - immunology
Influenza virus
Molecular Sequence Data
Neutralization Tests
Oxidative folding
Protein Folding
Protein Structure, Tertiary
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Summary: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.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2009.03.106