1. Recombinant haemagglutinin protein of highly pathogenic avian influenza A (H5N1) virus expressed in Pichia pastoris elicits a neutralizing antibody response in mice
- Author
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Sunil K. Lal, Dev Chandran, Chandrani Chakravarty, Lakshmi Narasu Mangamoori, Santhakumar Ponsekaran, Subathra Murugan, Lingaraja Kannivel, and Srinivasan Villuppanoor Alwar
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Gene Expression ,Hemagglutinin Glycoproteins, Influenza Virus ,Biology ,Antibodies, Viral ,medicine.disease_cause ,Pichia ,Poultry ,Pichia pastoris ,law.invention ,Mice ,Affinity chromatography ,Antigen ,Western blot ,law ,Virology ,medicine ,Animals ,Neutralizing antibody ,Vaccines, Synthetic ,Influenza A Virus, H5N1 Subtype ,medicine.diagnostic_test ,Immunogenicity ,Vaccination ,virus diseases ,biology.organism_classification ,Antibodies, Neutralizing ,Recombinant Proteins ,Influenza A virus subtype H5N1 ,Influenza Vaccines ,Influenza in Birds ,biology.protein ,Recombinant DNA ,Electrophoresis, Polyacrylamide Gel ,Female - 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.
- Published
- 2013
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