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The current live vaccine strain (LVS) vaccine poses safety and virulence concerns, advocating the need for a safer alternative. In this study, we demonstrated a combination of antigens FopA, Tul4, SucB, and DnaK conjugated to biopolymer particles (BPs) as a multivalent subunit vaccine that not only induces LVS and SchuS4-specific antibody titers but also confers protective immunity with differential cytokine profiles upon LVS challenge. This multivalent vaccine conferred 80% survival in mice, with antibodies of opsonizing ability and rapid recovery from weight loss. The production of these biopolymer conjugate vaccines was achieved in one step inside engineered Escherichia coli and is not only cost-effective but also leads to thermostable formulations, eliminating the need for cold-chain transport and storage. E. coli fermentation-based production avoids the need for high-containment facilities. Collectively, the FopA-BP-Tul4, SucB-BP, and Dnak-BP combined formulations hold the promise for the development of a robust subunit vaccine for the prevention of tularemia.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00694","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T16:01:45Z","timestamp":1789488105000},"page":"3543-3558","source":"Crossref","is-referenced-by-count":1,"title":["Biopolymer-Antigen\nConjugate Vaccines for Prevention\nof Tularemia"],"prefix":"10.1021","volume":"12","author":[{"given":"Gayathri","family":"Sam","sequence":"first","affiliation":[{"name":"Griffith University , , , ,","place":["Brisbane, Queensland, Australia, 4111"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8121-8463","authenticated-orcid":true,"given":"Charles T.","family":"Spencer","sequence":"additional","affiliation":[{"name":"University of Texas at El Paso , , , ,","place":["El Paso, Texas, United States, 79968"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3908-8903","authenticated-orcid":true,"given":"Bernd H. A.","family":"Rehm","sequence":"additional","affiliation":[{"name":"Griffith University , , , ,","place":["Brisbane, Queensland, Australia, 4111"]},{"name":"Griffith University , , , ,","place":["Gold Coast, Queensland, Australia, 4222"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,15]]},"reference":[{"key":"2026100909104530400_cit1","doi-asserted-by":"publisher","first-page":"1549343","DOI":"10.3389\/fmicb.2025.1549343","article-title":"The Immune Response to Francisella\ntularensis","volume":"16","author":"Barthels","year":"2025","journal-title":"Front. Microbiol."},{"issue":"1","key":"2026100909104530400_cit2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/01652176.2023.2277753","article-title":"Tularemia - A Re-emerging Disease with Growing Concern","volume":"43","author":"Sharma","year":"2023","journal-title":"Vet. Q."},{"issue":"Supplement_1","key":"2026100909104530400_cit3","doi-asserted-by":"publisher","first-page":"S1","DOI":"10.1093\/cid\/ciad681","article-title":"Tularemia: A Storied History, an Ongoing Threat","volume":"78","author":"Nelson","year":"2024","journal-title":"Clin. Infect. Dis."},{"issue":"Supplement_1","key":"2026100909104530400_cit4","doi-asserted-by":"publisher","first-page":"S29","DOI":"10.1093\/cid\/ciad689","article-title":"Tularemia Clinical Manifestations, Antimicrobial Treatment,\nand Outcomes: An Analysis of US Surveillance Data, 2006-2021","volume":"78","author":"Wu","year":"2024","journal-title":"Clin. Infect. 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Immunol."},{"key":"2026100909104530400_cit8","doi-asserted-by":"publisher","first-page":"154","DOI":"10.3389\/fcimb.2018.00154","article-title":"Live Attenuated\nTularemia Vaccines for Protection Against\nRespiratory Challenge With Virulent F. tularensis subsp. Tularensis","volume":"8","author":"Jia","year":"2018","journal-title":"Front. Cell. Infect. Microbiol."},{"issue":"8","key":"2026100909104530400_cit9","doi-asserted-by":"publisher","first-page":"3664","DOI":"10.1128\/IAI.00374-08","article-title":"Native Outer Membrane\nProteins Protect Mice Against\nPulmonary Challenge with Virulent Type A Francisella tularensis","volume":"76","author":"Huntley","year":"2008","journal-title":"Infect. Immun."},{"issue":"1","key":"2026100909104530400_cit10","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1093\/infdis\/135.1.55","article-title":"Immunization\nAgainst Tularemia: Analysis of the Effectiveness of Live Francisella\ntularensis Vaccine in Prevention of Laboratory-Acquired Tularemia","volume":"135","author":"Burke","year":"1977","journal-title":"J. Infect. Dis."},{"issue":"1","key":"2026100909104530400_cit11","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1093\/ajcp\/2.1.7","article-title":"Prophylactic\nVaccination Against Tularemia","volume":"2","author":"Foshay","year":"1932","journal-title":"Am. J. Clin.\nPathol."},{"issue":"10","key":"2026100909104530400_cit12","doi-asserted-by":"publisher","first-page":"1131","DOI":"10.2105\/AJPH.32.10.1131","article-title":"Vaccine Prophylaxis\nAgainst Tularemia in Man","volume":"32","author":"Foshay","year":"1942","journal-title":"Am. J. 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Microbiol."},{"issue":"10","key":"2026100909104530400_cit35","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0200213","article-title":"Protection Induced by a Francisella\ntularensis Subunit Vaccine Delivered by Glucan Particles","volume":"13","author":"Whelan","year":"2018","journal-title":"PLoS One"},{"issue":"9","key":"2026100909104530400_cit36","doi-asserted-by":"publisher","first-page":"1613","DOI":"10.4014\/jmb.1602.02022","article-title":"A Synthetic\nTul4 and FopA Peptide Cocktail of Francisella tularensis Induces Humoral\nand Cell-Mediated Immune Responses in Mice","volume":"26","author":"Oh","year":"2016","journal-title":"J. Microbiol. Biotechnol."},{"issue":"6","key":"2026100909104530400_cit37","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0130858","article-title":"Development of a\nMultivalent Subunit Vaccine against Tularemia Using Tobacco Mosaic\nVirus (TMV) Based Delivery System","volume":"10","author":"Banik","year":"2015","journal-title":"PLoS One"},{"issue":"Supplement_1","key":"2026100909104530400_cit38","doi-asserted-by":"publisher","first-page":"147.17","DOI":"10.4049\/jimmunol.198.Supp.147.17","article-title":"Development of a Multivalent Subunit\nVaccine against\nRespiratory Tularemia","volume":"198","author":"Mansour","year":"2017","journal-title":"J. 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