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The present study was conducted to purify and detect the immunogenicity of the fusion protein HtsA\u2009+\u2009FtsB of the iron transport lipoproteins HtsA and FtsB of <jats:italic>S. pyogenes<\/jats:italic>. The recombinant expression vector pBAD\u2010<jats:italic>htsA<\/jats:italic>\u2009+\u2009<jats:italic>ftsB<\/jats:italic> was successfully constructed, and the fusion protein HtsA\u2009+\u2009FtsB with a purity above 95% was successfully obtained. Western blot analysis confirmed that the HtsA\u2009+\u2009FtsB fusion protein had good antigenicity and could be specifically recognized by both HtsA antiserum and FtsB antisera, and the antibody specificity of the HtsA\u2009+\u2009FtsB fusion protein was good. ELISA results showed that IgG antibody levels were significantly increased in the HtsA\u2009+\u2009FtsB fusion protein immunization group compared to the PBS control group. Additionally, cytokine levels, including IL\u20102, IFN\u2010\u03b3, IL\u20104, IL\u20106, and IL\u201017A, were significantly elevated. Furthermore, the antiserum from HtsA\u2009+\u2009FtsB\u2010immunized mice enhanced the opsonophagocytic activity against <jats:italic>S. pyogenes<\/jats:italic>. The HtsA\u2009+\u2009FtsB fusion protein has strong immunogenicity and potential as a candidate vaccine for <jats:italic>S. pyogenes<\/jats:italic>.<\/jats:p>","DOI":"10.1111\/1348-0421.13217","type":"journal-article","created":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T10:34:44Z","timestamp":1744367684000},"page":"317-325","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Expression Purification and Immunogenicity Detection of HtsA\u2009+\u2009FtsB Fusion Protein From <i>Streptococcus pyogenes<\/i>"],"prefix":"10.1111","volume":"69","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-4547-5539","authenticated-orcid":false,"given":"Jing","family":"Li","sequence":"first","affiliation":[{"name":"College of Bioengineering Zhuhai Campus of Zunyi Medical University Zhuhai Guangdong 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