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Recombinant haemagglutinin protein of highly pathogenic avian influenza A (H5N1) virus expressed in Pichia pastoris elicits a neutralizing antibody response in mice

Authors :
Sunil K. Lal
Dev Chandran
Chandrani Chakravarty
Lakshmi Narasu Mangamoori
Santhakumar Ponsekaran
Subathra Murugan
Lingaraja Kannivel
Srinivasan Villuppanoor Alwar
Source :
Journal of Virological Methods. 187:20-25
Publication Year :
2013
Publisher :
Elsevier BV, 2013.

Abstract

Recombinant avian influenza vaccines offer several advantages over the conventional vaccines. In this study, the haemagglutinin (HA) gene of highly pathogenic avian influenza H5N1 was cloned and expressed as His tagged protein in methylotropic yeast Pichia pastoris. The expression of recombinant HA (rHA) protein was confirmed by SDS-PAGE and western blot analysis. The rHA protein was purified using Ni-NTA affinity chromatography under denaturing conditions and the functions of the protein was assessed by the haemagglutinin assay after refolding. The immunogenicity of the rHA was evaluated by immunizing four groups of mice with different payloads (2.5, 5.0, 10 and 25 μg) of purified rHA and the production of rHA specific antibodies were analysed by haemagglutinin inhibition assay (HI) and enzyme-linked immunosorbent assay (ELISA). An antigen specific immune response was observed against rHA indicating that the rHA antigen could be used as a vaccine candidate against avian influenza. These results suggest that this strategy would pave the way for the development of rapid and cost effective method for the production of an avian influenza vaccine.

Details

ISSN :
01660934
Volume :
187
Database :
OpenAIRE
Journal :
Journal of Virological Methods
Accession number :
edsair.doi.dedup.....363181ccfe9d8d3924062368e77beed4
Full Text :
https://doi.org/10.1016/j.jviromet.2012.07.026