{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T07:22:37Z","timestamp":1777706557310,"version":"3.51.4"},"reference-count":128,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T00:00:00Z","timestamp":1753401600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Monkeypox virus (MPXV) has caused 148,892 confirmed cases and 341 deaths from 137 countries worldwide, as reported by the World Health Organization (WHO), highlighting the urgent need for effective vaccines to prevent the spread of MPXV. Traditional vaccine development is low-throughput, expensive, time consuming, and susceptible to reversion to virulence. Alternatively, a reverse vaccinology approach offers a rapid, efficient, and safer alternative for MPXV vaccine design. Here, MPXV proteins associated with viral infection were analyzed for immunogenic epitopes to design multi-epitope vaccines based on B-cell, CD4+, and CD8+ epitopes. Epitopes were selected based on allergenicity, antigenicity, and toxicity parameters. The prioritized epitopes were then combined via peptide linkers and N-terminally fused to various protein adjuvants, including PADRE, beta-defensin 3, 50S ribosomal protein L7\/12, RS-09, and the cholera toxin B subunit (CTB). All vaccine constructs were computationally validated for physicochemical properties, antigenicity, allergenicity, safety, solubility, and structural stability. The three-dimensional structure of the selected construct was also predicted. Moreover, molecular docking and molecular dynamics (MD) simulations between the vaccine and the TLR-4 immune receptor demonstrated a strong and stable interaction. The vaccine construct was codon-optimized for high expression in the E. coli and was finally cloned in silico into the pET21a (+) vector. Collectively, these results could represent innovative tools for vaccine formulation against MPXV and be transformative for other infectious diseases.<\/jats:p>","DOI":"10.3390\/ijms26157210","type":"journal-article","created":{"date-parts":[[2025,7,25]],"date-time":"2025-07-25T12:56:43Z","timestamp":1753448203000},"page":"7210","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Development of a Broad-Spectrum Pan-Mpox Vaccine via Immunoinformatic Approaches"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-2736-3007","authenticated-orcid":false,"given":"Japigorn","family":"Puagsopa","sequence":"first","affiliation":[{"name":"Department of Physiology and Aging, College of Medicine, University of Florida, Gainesville, FL 32610, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Panuwid","family":"Jumpalee","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Science, King Mongkut\u2019s Institute of Technology Ladkrabang, Bangkok 10520, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9046-3239","authenticated-orcid":false,"given":"Sittichoke","family":"Dechanun","sequence":"additional","affiliation":[{"name":"Department of Bioengineering BioScience Research Collaborative, Rice University, Houston, TX 77005, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sukanya","family":"Choengchalad","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Science, King Mongkut\u2019s Institute of Technology Ladkrabang, Bangkok 10520, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pana","family":"Lohasupthawee","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Science, King Mongkut\u2019s Institute of Technology Ladkrabang, Bangkok 10520, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9126-0649","authenticated-orcid":false,"given":"Thanawat","family":"Sutjaritvorakul","sequence":"additional","affiliation":[{"name":"Faculty of Science and Technology, Pathumwan Institute of Technology, Bangkok 10330, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0222-3713","authenticated-orcid":false,"given":"Bunyarit","family":"Meksiriporn","sequence":"additional","affiliation":[{"name":"Department of Biology, School of Science, King Mongkut\u2019s Institute of Technology Ladkrabang, Bangkok 10520, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1528","DOI":"10.1097\/JS9.0000000000000077","article-title":"Efficacy of Smallpox Approved Tecovirimat (Tpoxx) Drug against Monkeypox: Current Update and Futuristic Prospects","volume":"109","author":"Das","year":"2023","journal-title":"Int. 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