Giuseppe
Emeritus
Biochem Biophys Res Commun. 2009 Mar 23. [Epub ahead of print]
Immunological Study of HA1 Domain of Heamagglutinin of Influenza H5N1 Virus.
Chiu FF, Venkatesan N, Wu CR, Chou AH, Chen HW, Lian SP, Liu SJ, Huang CC, Lian WC, Chong P, Leng CH. - Vaccine Research and Development Center, National Health Research Institutes; Zhunan, Miaoli 350, Taiwan, ROC.
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.
PMID: 19324009 [PubMed - as supplied by publishe
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Immunological Study of HA1 Domain of Heamagglutinin of Influenza H5N1 Virus.
Chiu FF, Venkatesan N, Wu CR, Chou AH, Chen HW, Lian SP, Liu SJ, Huang CC, Lian WC, Chong P, Leng CH. - Vaccine Research and Development Center, National Health Research Institutes; Zhunan, Miaoli 350, Taiwan, ROC.
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.
PMID: 19324009 [PubMed - as supplied by publishe
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