Evaluation of antibody response in mice against avian influenza A (H5N1) strain neuraminidase expressed in yeast Pichia pastoris.
Avian influenza has raised many apprehension in the recent years because of its potential transmitability to humans. With the increasing emergence of drug-resistant avian influenza strains, development of potential vaccines are imperative to manage this disease. Two structural antigens, haemagglutinin and neuraminidase, have been the target candidates for the development of subunit vaccine against influenza. In an effort to develop a faster and economically beneficial vaccine, the neuraminidase gene of a highly pathogenic avian influenza isolate was cloned and expressed in the methylotrophic yeast Pichia pastoris. The recombinant neuraminidase (rNA) antigen was purified, and its bioactivity was analysed. The rNA was found to be functional, as determined by the neuraminidase assay. Four groups of mice were immunized with different concentrations of purified rNA antigen, which were adjuvanted with aluminium hydroxide. The immune response against rNA was analysed by enzyme-linked immunosorbent assay (ELISA) and neuraminidase inhibition assay. The mice groups immunized with 25 (mu) g and 10 (mu) g of antigen had a significant immune response against rNA. This method can be utilized for faster and cost-effective development of vaccines for a circulating and newer strain of avian influenza, and would aid in combating the disease in a pandemic situation, in which production time matters greatly.
Centre for Biotechnology, Institute of Science and Technology, Jawaharlal Nehru Technological University, Hyderabad, India.
Murugan
M
Subathra
Ponsekaran
P
Santhakumar
Mangamoori Lakshmi
ML
Narasu
Syed Sultan
SS
Beevi
Sunil K
SK
Lal
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D053124:Influenza A Virus, H5N1 Subtype / Q000201:enzymology / Q000276:immunology*; D007252:Influenza Vaccines; D051379:Mice; D009439:Neuraminidase / Q000235:genetics / Q000276:immunology*; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007252:Influenza Vaccines; D011994:Recombinant Proteins; D009439:Neuraminidase