Immunological study of HA1 domain of hemagglutinin of influenza H5N1 virus.
Biochem Biophys Res Commun
; 383(1): 27-31, 2009 May 22.
Article
em En
| MEDLINE
| ID: mdl-19324009
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.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Vacinas contra Influenza
/
Glicoproteínas de Hemaglutininação de Vírus da Influenza
/
Virus da Influenza A Subtipo H5N1
Limite:
Humans
Idioma:
En
Revista:
Biochem Biophys Res Commun
Ano de publicação:
2009
Tipo de documento:
Article