{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T02:27:11Z","timestamp":1777516031041,"version":"3.51.4"},"reference-count":60,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2008,9,5]],"date-time":"2008-09-05T00:00:00Z","timestamp":1220572800000},"content-version":"vor","delay-in-days":1343,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2005,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A two\u2010component recombinant fusion protein antigen was re\u2010engineered and tested as a medical counter measure against the possible biological threat of aerosolized <jats:italic>Yersinia pestis<\/jats:italic>. The active component of the proposed subunit vaccine combines the F1 capsular protein and V virulence antigen of <jats:italic>Y. pestis<\/jats:italic> and improves upon the design of an earlier histidine\u2010tagged fusion protein. In the current study, different production strains were screened for suitable expression and a purification process was optimized to isolate an F1\u2010V fusion protein absent extraneous coding sequences. Soluble F1\u2010V protein was isolated to 99% purity by sequential liquid chromatography including capture and refolding of urea\u2010denatured protein via anion exchange, followed by hydrophobic interaction, concentration, and then transfer into buffered saline for direct use after frozen storage. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing, confirming a purified product of 477 amino acids and removal of the N\u2010terminal methionine. Purity, quality, and higher\u2010order structure were compared between lots using RP\u2010HPLC, intrinsic fluorescence, CD spectroscopy, and multi\u2010angle light scattering spectroscopy, all of which indicated a consistent and properly folded product. As formulated with aluminum hydroxide adjuvant and administered in a single subcutaneous dose, this new F1\u2010V protein also protected mice from wild\u2010type and non\u2010encapsulated <jats:italic>Y. pestis<\/jats:italic> challenge strains, modeling prophylaxis against pneumonic and bubonic plague. These findings confirm that the fusion protein architecture provides superior protection over the former licensed product, establish a foundation from which to create a robust production process, and set forth assays for the development of F1\u2010V as the active pharmaceutical ingredient of the next plague vaccine.<\/jats:p>","DOI":"10.1021\/bp050098r","type":"journal-article","created":{"date-parts":[[2005,10,7]],"date-time":"2005-10-07T00:20:03Z","timestamp":1128644403000},"page":"1490-1510","source":"Crossref","is-referenced-by-count":82,"title":["Design and Testing for a Nontagged F1\u2010V Fusion Protein as Vaccine Antigen against Bubonic and Pneumonic Plague"],"prefix":"10.1002","volume":"21","author":[{"given":"Bradford S.","family":"Powell","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerard P.","family":"Andrews","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey T.","family":"Enama","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Scott","family":"Jendrek","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chris","family":"Bolt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia","family":"Worsham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey K.","family":"Pullen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wilson","family":"Ribot","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harry","family":"Hines","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leonard","family":"Smith","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David G.","family":"Heath","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey J.","family":"Adamovicz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2008,9,5]]},"reference":[{"key":"e_1_2_7_2_2","unstructured":"Abbreviations used: Ab antibody; AEX anion exchange chromatography; CEX cation exchange chromatography; CIEF capillary isoelectric focusing; des\u2010Met without the N\u2010terminal translation initiator methionine; DO2 dissolved oxygen; DPBS Dulbecco's phosphate\u2010buffered saline without Ca2+or Mg2+; F1\u2010V fusion protein comprisingY. pestisfraction 1 capsule joined to V antigen; FDA Food and Drug Administration; GMP good manufacturing practices; GMT geometric mean titer; GSD geometric standard deviation; HPLC high\u2010pressure liquid chromatography; IEP isoelectric point; IPTG isopropyl\u2010\u03b2\u2010d\u2010thiogalactoside; KWC killed whole cell; LD50 50% lethal dose; MALDI\u2010TOF\u2010MS matrix\u2010assisted laser desorption ionization time\u2010of\u2010flight mass spectrometry; Met methionine;Mr relative molecular weight; MST mean survival time as measured in days;m\/z mass\u2010to\u2010charge; OD405 optical density at 405 nm wavelength; PCR polymerase chain reaction; RFLP restriction fragment length polymorphism; RP\u2010HPLC reversed\u2010phase high\u2010pressure liquid chromatography; SC subcutaneous; SE standard error; SEC size exclusion chromatography; SEC\u2010MALS size exclusion chromatography coupled to multi\u2010angle light scattering spectroscopy; TetR tetracycline resistance; USAMRIID United States Army Medical Research Institute of Infectious Diseases; YOP Yersiniaouter protein."},{"key":"e_1_2_7_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0408026101"},{"key":"e_1_2_7_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.4.1516-1519.2000"},{"key":"e_1_2_7_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/0-306-48416-1_11"},{"key":"e_1_2_7_6_2","doi-asserted-by":"publisher","DOI":"10.1007\/s001090050148"},{"key":"e_1_2_7_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.10.1.35"},{"key":"e_1_2_7_8_2","unstructured":"Plague WHO Fact Sheet No. 267 Revised February 2005;http:\/\/www.who.int\/mediacentre\/factsheets\/fs267\/en\/."},{"key":"e_1_2_7_9_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199709043371004"},{"key":"e_1_2_7_10_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0701.010106"},{"key":"e_1_2_7_11_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.283.17.2281"},{"key":"e_1_2_7_12_2","first-page":"479","volume-title":"Medical Aspects of Chemical & Biological Warfare, Part I","author":"McGovern T.","year":"1997"},{"key":"e_1_2_7_13_2","doi-asserted-by":"publisher","DOI":"10.1001\/archderm.135.3.311"},{"key":"e_1_2_7_14_2","unstructured":"Davis J.;Johnson\u2010Winegar A.The anthrax terror: DOD's number\u2010one biological threat.Aerospace Power J.2000 xiv 15\u201329."},{"key":"e_1_2_7_15_2","volume-title":"Biohazard: The Chilling True Story of the Largest Covert Biological Weapons Program in the World","author":"Alibek K.","year":"1999"},{"key":"e_1_2_7_16_2","doi-asserted-by":"crossref","first-page":"2180","DOI":"10.1128\/iai.64.6.2180-2187.1996","article-title":"Fraction 1 capsular antigen (F1) purification from Yersinia pestis CO92 and from an Escherichia coli recombinant strain and efficacy against lethal plague challenge","volume":"64","author":"Andrews G. P.","year":"1996","journal-title":"Infect. Immun."},{"key":"e_1_2_7_17_2","doi-asserted-by":"crossref","first-page":"793","DOI":"10.4269\/ajtmh.1998.58.793","article-title":"Short\u2010 and long\u2010term efficacy of single\u2010dose subunit vaccines against Yersinia pestis in mice","volume":"58","author":"Anderson G. W.","year":"1998","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"e_1_2_7_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(98)80110-2"},{"key":"e_1_2_7_19_2","doi-asserted-by":"crossref","first-page":"4580","DOI":"10.1128\/iai.64.11.4580-4585.1996","article-title":"Recombinant V antigen protects mice against pneumonic and bubonic plague caused by F1\u2010capsule\u2010positive and \u2010negative strains of Yersinia pestis","volume":"64","author":"Anderson G. W.","year":"1996","journal-title":"Infect. Immun."},{"key":"e_1_2_7_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.4.1922-1928.1999"},{"key":"e_1_2_7_21_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2000.tb09086.x"},{"key":"e_1_2_7_22_2","first-page":"127","article-title":"TheV antigen of yersiniae: an overview","volume":"12","author":"Brubaker R. R.","year":"1991","journal-title":"Contrib. Microbiol. Immunol."},{"key":"e_1_2_7_23_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20020908"},{"key":"e_1_2_7_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(96)00303-9"},{"key":"e_1_2_7_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(88)90394-0"},{"key":"e_1_2_7_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.171.10.5646-5653.1989"},{"key":"e_1_2_7_27_2","doi-asserted-by":"publisher","DOI":"10.1006\/prep.1998.0910"},{"key":"e_1_2_7_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(80)90165-7"},{"key":"e_1_2_7_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(99)01540-6"},{"key":"e_1_2_7_30_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1026376403468"},{"key":"e_1_2_7_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2004.11.001"},{"key":"e_1_2_7_32_2","first-page":"116","volume-title":"Guide for the Care and Use of Laboratory Animals","year":"1996"},{"key":"e_1_2_7_33_2","unstructured":"Bovine Spongiform Encephalopathy (BSE) U.S. Food and Drug Administration Center for Biologics Evaluation and Research and references therein http:\/\/www.fda.gov\/cber\/bse\/bse.htm accessed 31 March 2005."},{"key":"e_1_2_7_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/0734-9750(94)90292-5"},{"key":"e_1_2_7_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)36503-2"},{"key":"e_1_2_7_36_2","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1128\/AEM.64.3.982-991.1998","article-title":"Fermentation, purification, and characterization of protective antigen from a recombinant, avirulent strain of Bacillus anthracis","volume":"64","author":"Farchaus J. W.","year":"1998","journal-title":"Appl. Environ. Microbiol."},{"key":"e_1_2_7_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-4347(94)00078-6"},{"key":"e_1_2_7_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(96)01511-6"},{"key":"e_1_2_7_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2004.08.009"},{"key":"e_1_2_7_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2003.08.023"},{"key":"e_1_2_7_41_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/129.Supplement_1.S19"},{"key":"e_1_2_7_42_2","first-page":"653","article-title":"Effectiveness of live or killed plague vaccines in man","volume":"42","author":"Meyer K.","year":"1970","journal-title":"Bull. WHO"},{"key":"e_1_2_7_43_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.1995.tb00196.x"},{"key":"e_1_2_7_44_2","unstructured":"Fed. Regist. 2004 29 249 Biological products; bacterial vaccines and toxoids implementation of efficacy review"},{"key":"e_1_2_7_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(00)00159-6"},{"key":"e_1_2_7_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(01)00453-4"},{"key":"e_1_2_7_47_2","doi-asserted-by":"crossref","unstructured":"Fed. Regist. 2002 29 105","DOI":"10.7146\/nys.v29i29.13427"},{"key":"e_1_2_7_48_2","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1128\/IAI.67.3.1533-1537.1999","article-title":"Protective efficacy of recombinant Yersinia outer proteins against bubonic plague caused by encapsulated and non\u2010encapsulated Yersinia pestis","volume":"67","author":"Andrews G. P.","year":"1999","journal-title":"Infect. Immun."},{"key":"e_1_2_7_49_2","first-page":"29","volume-title":"Protein Purification Protocols","author":"Cutler P.","year":"2004"},{"key":"e_1_2_7_50_2","unstructured":"Protein structure initiative protein production workshop. National Institute of General Medical Sciences Mar 7\u2013;8 2002; available athttp:\/\/www.nigms.nih.gov\/news\/reports\/protein_production.html#session1."},{"key":"e_1_2_7_51_2","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/5.6.593"},{"key":"e_1_2_7_52_2","doi-asserted-by":"publisher","DOI":"10.1006\/prep.2001.1416"},{"key":"e_1_2_7_53_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.2.751-757.1987"},{"key":"e_1_2_7_54_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2249.1999.00859.x"},{"key":"e_1_2_7_55_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0808.010516"},{"key":"e_1_2_7_56_2","unstructured":"Anderson G. W.;Worsham P. L.;Andrews G. P.;Bolt C. R.;Welkos S. L.;Friedlander A. M.;Burans J. P.Passive immunization with monoclonal antibodies against the F1 antigen ofYersinia pestisprotects mice from fatal bubonic and pneumonic plague. Conference poster at the American Society of Tropical Medicine & Hygiene San Antonio TX 20 November 1995."},{"key":"e_1_2_7_57_2","unstructured":"Andrews G. P.;Bolt C. R.;Pitt L. M.;Adamovicz J. J.Passive protection in mice from fatal pneumonic plague by antibody raised against a recombinantYersinia pestisF1\u2010V fusion protein vaccine candidate. Poster at American Society for Microbiology 104th General Meeting 2004."},{"key":"e_1_2_7_58_2","doi-asserted-by":"crossref","first-page":"2226","DOI":"10.4049\/jimmunol.133.4.2226","article-title":"Roles of V antigen in promoting virulence and immunity in yersiniae","volume":"133","author":"Une T.","year":"1984","journal-title":"J. Immunol."},{"key":"e_1_2_7_59_2","first-page":"299","article-title":"Studies on the contribution of the F1 capsule\u2010associated plasmid pFra to the virulence of Yersinia pestis","volume":"13","author":"Welkos S. L.","year":"1995","journal-title":"Contrib. Microbiol. Immunol."},{"key":"e_1_2_7_60_2","first-page":"325","article-title":"Construction of defined F1 negative mutants of virulent Yersinia pestis","volume":"13","author":"Worsham P. L.","year":"1995","journal-title":"Contrib. Microbiol. Immunol."},{"key":"e_1_2_7_61_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.71.4.2234-2238.2003"}],"container-title":["Biotechnology Progress"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1021%2Fbp050098r","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/aiche.onlinelibrary.wiley.com\/doi\/pdf\/10.1021\/bp050098r","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T21:16:35Z","timestamp":1760044595000},"score":1,"resource":{"primary":{"URL":"https:\/\/aiche.onlinelibrary.wiley.com\/doi\/10.1021\/bp050098r"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,1]]},"references-count":60,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2005,1]]}},"alternative-id":["10.1021\/bp050098r"],"URL":"https:\/\/doi.org\/10.1021\/bp050098r","archive":["Portico"],"relation":{},"ISSN":["8756-7938","1520-6033"],"issn-type":[{"value":"8756-7938","type":"print"},{"value":"1520-6033","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,1]]}}}