In-silico analysis and design of a chimeric protein containing B-cell epitopes of proteins for the control of Mpox

Abstract

Mpox, caused by the Monkey pox virus (MPXV), remains a public health challenge due to its genetic diversity with multiple clades of varying virulence and geographic spread. The current live-attenuated vaccine, JYNNEOS (Modified Vaccinia Ankara-based), primarily targets clade II and offers partial cross-protection but carries a risk of reversion to virulence. It relies on vaccinia virus proteins like A33R, B5R, L1R, and A27L, which share homology with MPXV but do not cover the full antigenic diversity. Limitations include clade specificity, safety concerns, and replicative potential, restricting broad application. To address these gaps, this study used in silico techniques to design a multi-epitope chimeric protein vaccine incorporating immunodominant epitopes from five MPXV proteins A35R, C19L, E8L, D10L, and M1R—key in viral infection and immune response. Epitopes are the part of a protein, usually a small peptide sequence, that is recognized by the immune system and triggers an immune response. Fifteen shortlisted epitopes were mapped and combined with the APPHALS adjuvant and linkers, forming a 533-amino acid vaccine chimera with a molecular weight of 57,204.85 Da. The vaccine was predicted to be highly immunogenic, non-allergenic, non-toxic, antigenic, hydrophilic, and stable, supporting its safety. The proposed vaccine candidate leverages conserved epitopes shared across different clades, enhancing potential cross-protection and broadening the immune coverage crucial for effective global Mpox control. The study highlights the promise of in silico vaccine design in developing broad-spectrum, safe, and effective Mpox vaccines. Future work should include experimental validation through in vitro and in vivo studies in relevant models to confirm immunogenicity, efficacy, and safety toward clinical development.