Sourabh Suran, University of Saskatchewan

Zahed Khatooni, Bryce Warner

Fighting & Responding

Objectives The study aims to identify B and T cell epitopes from new world hantaviruses, design and develop a structurally validated, multi-epitope vaccine candidate using an integrated structural biology approach. Methods: A stepwise immunoinformatic pipeline where the complete Andes virus glycoprotein 3D conformation was simulated and evaluated. The potential B-cell and T-cell epitopes were evaluated using BediPred and Discotope tools. Over 1,000 overlapping nine-mer peptides from the predicted conformation were generated to cover possible T-cell epitopes comprehensively. The conformations of nine-mer peptides were individually modelled and compared to ensure structural integrity. Globally prevalent HLA class I and II allele sets were generated. To simulate the interaction of each peptide with each HLA allele, a large-scale cross-docking analysis was performed, and broad population coverage and immunodominant candidates were predicted. The top peptides, selected based on docking scores and binding energies, were incorporated into a multi-epitope construct integrating both B and T cell targets. Molecular dynamics simulations were carried out to evaluate the construct's structural stability, flexibility, and physicochemical properties followed by recombinational expression and structural-functional validation. Results and conclusion: Several conserved and highly immunogenic epitopes were identified throughout the glycoprotein complex and nucleocapsid from new world hantaviruses. Out of over 1000 modelled peptides, many demonstrated strong binding affinities with multiple HLA alleles, supported by stable and complementary docking poses. This study demonstrates a high-throughput, structure-based computational pipeline for rational epitope vaccine design. The approach effectively identified multiple promising B and T cell epitopes from the glycoprotein and nucleocapsid, to be assembled