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In view of the potential biological and practical benefits of a recombinant protein vaccine composed of a single fusion protein containing flagellin and antigen, we have evaluated the efficacy of a fusion protein composed of flagellin and two protective antigens of<jats:italic>Yersinia pestis<\/jats:italic>(F1 and V) in eliciting protection against respiratory challenge with<jats:italic>Y. pestis<\/jats:italic>. Flagellin-F1-V was produced and purified in high yield under good manufacturing practices conditions. The fusion protein retains full Toll-like receptor 5-stimulating activity in vitro. Using a prime-boost immunization protocol, we found that flagellin-F1-V elicits robust antigen-specific humoral immunity in mice and two species of nonhuman primates. Immune mice were fully protected against intranasal challenge with 150 mean tolerated doses of<jats:italic>Y. pestis<\/jats:italic>CO92. In immune mice, the bacteria were completely cleared within 3 days after challenge. Flagellin-F1-V exhibited full stability for at least 297 days at 4\u00b0C and at least 168 days at 25\u00b0C. At between 29 and 84 days at 37\u00b0C, the protein exhibited a loss of biological activity that appeared to be associated with a substantial change in protein diameter, possibly due to oligomerization. On the basis of our results, we believe that flagellin-F1-V is an outstanding candidate for evaluation in studies with humans.<\/jats:p>","DOI":"10.1128\/cvi.00333-08","type":"journal-article","created":{"date-parts":[[2008,11,6]],"date-time":"2008-11-06T01:54:22Z","timestamp":1225936462000},"page":"21-28","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":128,"title":["Flagellin-F1-V Fusion Protein Is an Effective Plague Vaccine in Mice and Two Species of Nonhuman Primates"],"prefix":"10.1128","volume":"16","author":[{"given":"Steven B.","family":"Mizel","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology"},{"name":"Department of Pediatrics"},{"name":"Section on Comparative Medicine"},{"name":"Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157"},{"name":"Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland 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