A novel multi-epitope-based peptide recombinant influenza A vaccine prototype utilizing neuraminidase and hemagglutinin surface proteins: From in silico to preliminary study.

Neuraminidase and hemagglutinin serve as the crucial surface proteins of influenza viruses. Hemagglutinin, as a variable surface protein, is indispensable for vaccine development. Therefore, Neuraminidase must not be overlooked in the formulation of the recombinant vaccine prototype, which may serve as a candidate for designing a multi-epitope recombinant vaccine using immunoinformatics. Our study involves immunoinformatic screening and analysis to develop a recombinant multi-epitope vaccine prototype comprising immunodominant and conserved epitopes from influenza hemagglutinin and neuraminidase. Predicted B-cell and T-cell epitopes target a wide allele population. A 199-amino acid construct integrates MHCI and MHCII for both mouse and human hosts, connected by rigid and flexible linkers. Molecular docking findings suggest that this multi-epitope structure could activate TLR3, TLR7, and TLR8, thereby prompting protective immune responses. B-cell epitopes mediate adaptive immune responses by facilitating antigen recognition and memory formation Furthermore, the designed construct underwent in silico cloning of the vaccine prototype candidate in pET21a as a prokaryotic expression vector, followed by evaluation and exploration. It underwent characterization for physicochemical attributes, allergenicity, toxicity, and antigenicity. Validation through dynamic simulation confirms the stability of the construct. This pioneering immunoinformatic study proposes the potential of a recombinant protein vaccine prototype centered around neuraminidase and hemagglutinin immunodominant epitopes to elicit immune responses against a broad spectrum of viruses. Additionally, this vaccine prototype has been evaluated through both in silico and in vitro studies.