Development of bacterially expressed recombinant HA1 based vaccine and its immunogenic evaluation in mice model against H1N1 influenza virus

Abstract

Influenza viruses are contagious evolved as major respiratory pathogen. The present research work was carried out with the objectives of in silico study, cloning and newlineexpression of HA1 gene of central Indian H1N1pdm 09 virus and inoculation of expressed protein mice model. In silico analysis of already submitted HA sequences revealed five genogroups (2, 3, 4, 6 (6A, 6B, 6C) and 7 genogroups) evolved between 2009 to 2019 in central India. In this study 100 throat swab samples were collected, out of that 48 tested positive for Inf A (H1N1)pdm 09. 14 samples HA1 gene amplified by 2 Step PCR. 5 samples were successfully sequenced. The sequences of 5 samples after analysis revealed clustering in genogroup 6B. There were many mutations noted after alignment with Michigan Vaccine newlinestrain. A Central Indian H1N1 sample with HA gene sequence accession number MZ735580.1 Influenza A virus (A/Bhopal/AS_1008/2019(H1N1)) was used for cloning into prokaryotic expression vector, transformed into E.coli Bl-21 (DE3), and protein was expressed and purified. The purified protein along with adjuvant was inoculated in Swiss Albino mice and serological testing was done by ELISA. The antibody titre enhanced after second booster dose of rHA1 protein. The results indicated successful antigen preparation and antibody generation in mice. newline