Integrative reverse vaccinology and computational modeling for the rational design of a broadly immunogenic multi-epitope vaccine against Marburg virus infection.

Marburg virus (MARV), a member of the Filoviridae family, causes severe hemorrhagic fever in humans with case fatality rates exceeding 90%, and currently, no approved vaccines or therapeutics are available. To address this urgent need, we employed comprehensive immunoinformatics and computational approaches to design a multi-epitope subunit vaccine (MESV) capable of eliciting robust immune responses. Highly antigenic, non-allergenic, and non-toxic cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and B-cell epitopes derived from MARV glycoprotein (GP) and nucleoprotein (NP) were selected and assembled using appropriate linkers and adjuvant sequences. The designed vaccine construct exhibited favorable physicochemical characteristics, structural stability, and strong immunogenic potential. Solubility analysis predicted a score of 0.835, while structural validation revealed an ERRAT score of 94.11%, with 89.4% of residues located in the most favored regions of the Ramachandran plot. The ProSA analysis yielded a Z-score of - 5.31, confirming the reliability of the modeled structure. Molecular docking studies demonstrated strong interactions between the vaccine construct and Toll-like receptor 7, while molecular dynamics simulations confirmed the stability of the docked complex. Codon optimization and in silico cloning indicated efficient expression potential in Escherichia coli, with a codon adaptation index (CAI) of 0.9805 and GC content of 55.39%. Furthermore, immune simulations predicted a robust and sustained immune response. These computational findings suggest that the designed MESV is a promising vaccine candidate for MARV and warrants further experimental validation through in vitro and in vivo studies.