An immuno-informatics approach for annotation of hypothetical proteins and multi-epitope vaccine designed against the Mpox virus

Abstract

A worrying new outbreak of Monkeypox (Mpox) in humans is caused by the Mpox virus (MpoxV). The pathogen has roughly 28 hypothetical proteins of unknown structure, function, and pathogenicity. Using reliable bioinformatics tools, we attempted to analyze the MpoxV genome, identify the role of hypothetical proteins (HPs), and design a potential candidate vaccine. Out of 28, we identified seven hypothetical proteins using multi-server validation with high confidence for the occurrence of conserved domains. Their physical, chemical, and functional characterizations, including molecular weight, theoretical isoelectric point, 3D structures, GRAVY value, subcellular localization, functional motifs, antigenicity, and virulence factors, were performed. We predicted possible cytotoxic T cell (CTL), helper T cell (HTL) and linear and conformational B cell epitopes, which were combined in a 219 amino acid multiepitope vaccine with human β defensin as a linker. This multi-epitopic vaccine was structurally modelled and docked with toll-like receptor-3 (TLR-3). The dynamical stability of the vaccine-TLR-3 docked complexes exhibited stable interactions based on RMSD and RMSF tests. Additionally, the modelled vaccine was cloned in-silico in an E. coli host to check the appropriate expression of the final vaccine built. Our results might conform to an immunogenic and safe vaccine, which would require further experimental validation.

Communicated by Ramaswamy H. Sarma

Keywords:

• Mpox
• hypothetical protein
• multiepitope vaccine design
• molecular docking
• molecular dynamics simulation

Acknowledgments

The authors acknowledge the High Performance Cluster, Indian Institute of Technology, Delhi for providing the computational resources. The authors are grateful to Dr. Adnan Mannan for his suggestions and critical reviews. We are especially thankful to Md. Muzahidul Islam Ezaj, Sadia Binte Shams and Sreejoya Lala for their praiseworthy assistance.

Author contributions

Conceptualization, MHA; Data curation, AH, FA, GS, MHA, VG; Investigation, AKD, AR, MFMM; Methodology, AR, MFMM; Resources, FA, PA, SMA; Software, AH, AKD, GS, MHA, NSNS; Supervision, FA, VG; Validation, GS, VG; Visualization, FA, GS; Writing—original draft, EHF, JS, MAL, MHA, KHS; Writing—review & editing, GS, MHA, NSNS, SMA, PA, and VG.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data generated or analyzed during this study are included in this published article [and its supplementary information files]. Data used and/or analyzed during the current study is also available from the corresponding author upon reasonable request.

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.