Work

Protocol for designing a peptide-based multi-epitope vaccine targeting monkeypox using reverse vaccine technology

Year: 2025

Type: article

Abstract: Reverse vaccine technology, supported by advancements in immunoinformatics, facilitates the development of multi-epitope vaccines for rapidly evolving pathogens, thereby strengthening the immune defense. Here, we present a protocol for a peptide-based multi-epitope vaccine targeting monkeypox virus (MPXV) using an open-source approach. We describe steps for evaluating physicochemical properties and allergenicity. We then detail procedures for validating pattern recognition receptor (PRR)-binding affinity and stable major histocompatibility complex (MHC) I/II presentation. Molecular dynamics (MD) simulations confirm immune receptor interactions, enhancing vaccine stability. For complete details on the use and execution of this protocol, please refer to Kaur et al. 1 • Steps for predicting specific epitopes for B cells, T cells, and IFN-gamma • Instructions for the construction of 3D vaccine structure and its validation • Steps for molecular docking and MD simulation with PRRs • Guidelines on in silico cloning and immune simulation Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Reverse vaccine technology, supported by advancements in immunoinformatics, facilitates the development of multi-epitope vaccines for rapidly evolving pathogens, thereby strengthening the immune defense. Here, we present a protocol for a peptide-based multi-epitope vaccine targeting monkeypox virus (MPXV) using an open-source approach. We describe steps for evaluating physicochemical properties and allergenicity. We then detail procedures for validating pattern recognition receptor (PRR)-binding affinity and stable major histocompatibility complex (MHC) I/II presentation. Molecular dynamics (MD) simulations confirm immune receptor interactions, enhancing vaccine stability.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103671