In silico design of a multiepitope vaccine candidate targeting the monkeypox virus L1R protein via sequence-based immunoinformatics

Monkeypox virus (MPXV) is a zoonotic orthopoxvirus whose re-emergence poses a significant global health threat. The L1R protein, a highly conserved component of the intracellular mature virion (IMV) membrane, plays a critical role in viral entry and replication, making it a promising target for epitope-based vaccine design. In this study, an immunoinformatics-driven in silico approach was employed to design a structurally stable, non-toxic, and non-allergenic multi-epitope vaccine candidate derived from the MPXV L1R protein. Human protein homology screening using BLASTp confirmed the absence of significant similarity, thereby minimizing the risk of autoimmunity. Predicted cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and B-cell epitopes were linked using immunogenic linkers and adjuvanted with human β-defensin 3. The construct was evaluated for toxicity, allergenicity, three-dimensional structure, molecular docking with Toll-like receptors 2 (TLR2) and 4 (TLR4), structural stability, and immune response simulation. The optimized construct exhibited high predicted immunogenicity, safety, and structural robustness. Molecular docking analysis revealed strong binding affinity toward both TLR2 and TLR4, while immune simulations predicted robust activation of humoral and cellular immune responses. Collectively, these findings suggest that the proposed construct represents a promising next-generation vaccine candidate against orthopoxviruses and warrants further experimental validation.