Abstract
BACKGROUND: Monkeypox virus (MPXV) causes mpox, and vaccination is the most effective method of prevention. This project utilized immunoinformatics tools to design a multiepitope subunit vaccine against Mpox. MATERIALS AND METHODS: Seven MPXV antigens (B18R, B21R, D2L, E8L, N3R, I1L, and N2R) were selected based on a literature review, and their sequences were retrieved with the signal peptides and transmembrane domains omitted. B cell, T cell, MHC-I, and MHC-II epitopes were evaluated, followed by tertiary structure and homology modeling. The physicochemical properties of the construct were assessed, molecular docking was conducted with murine Toll-like receptors, and codon optimization was performed. RESULTS: Three sequences were excluded: B18R lacked appropriate epitopes, N2R was predicted to be an allergen by AllergenFP software, and I1L lacked antigenicity. The final vaccine construct incorporated segments from four genes, along with two adjuvants and two T-helper molecules linked by short sequences. The designed protein had a pI of 7.96, a molecular weight of approximately 16 kDa, an energy value of -723.06, a codon frequency distribution of 100, and a codon adaptation index of 0.88. The construct was subsequently cloned and inserted into a pET-28a(+) vector to generate a recombinant plasmid. CONCLUSIONS: Using immunoinformatics techniques, a novel multiepitope vaccine construct for MPXV was successfully designed.