{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:42:00Z","timestamp":1785436920964,"version":"3.56.0"},"posted":{"date-parts":[[2025,10,31]]},"group-title":"Immunology","reference-count":43,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T00:00:00Z","timestamp":1761868800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["#R01-AI125421-01A1"],"award-info":[{"award-number":["#R01-AI125421-01A1"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["#1RO1AI141229"],"award-info":[{"award-number":["#1RO1AI141229"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/0517h6h17","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,31]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Transferrin binding protein B (TbpB), an iron acquisition protein, has long been recognized as a promising vaccine candidate targeting the pathogenic Neisseria species, including\n                  <jats:italic>Neisseria meningitidis<\/jats:italic>\n                  , the cause of meningococcal disease, and\n                  <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                  , the cause of gonorrhea. A challenge to the development of this protein as a vaccine immunogen is the extent of antigenic variability it exhibits, which complicates the selection of a single variant to elicit a broadly cross-protective immune response. We have utilized structure-informed antigen engineering to develop a minimized version of TbpB consisting of the protein\u2019s carboxy-terminal lobe with its variable surface loops removed. Here, we reveal the effectiveness of this \u201cloopless C-lobe\u201d as an independent immunogen, with structural characterization and stability studies to demonstrate its integrity, and murine immunization and challenge studies that establish its ability to elicit robust protective efficacy by using\n                  <jats:italic>N. meningitidis<\/jats:italic>\n                  invasive infection and nasopharyngeal colonization models. The breadth of protection provided, as measured by both in vitro analysis and cross-protection mouse challenge studies, indicate that a single loopless C-lobe elicits a broadly cross-protective immune response against the diverse panel of meningococcal strains tested, and that the cross-reactivity is superior to that offered by the intact TbpB or the native C-lobe. Together, this study demonstrates the utility of structure-informed antigen engineering towards the development of broadly efficacious protein-based vaccines.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    Surface-exposed proteins on bacterial pathogens are enticing candidate vaccine targets, however their exposure to the immune system frequently leads to high levels of antigenic variation, a factor that complicates the development of broadly protective vaccines. Here, we undertake an antigen engineering approach to develop a minimized version of a surface lipoprotein, transferrin binding protein B, where variable regions of the protein have been removed to focus the immune response to conserved regions of this antigen. We combine structural studies and mouse infection models of\n                    <jats:italic>Neisseria meningitidis<\/jats:italic>\n                    , the cause of meningococcal disease, and\n                    <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                    , the causative agent of gonorrhea, to reveal that our strategic minimizing of the protein immunogen focuses the immune response to extend the resulting breadth of cross-reactivity and cross-protection.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.10.30.685603","type":"posted-content","created":{"date-parts":[[2025,11,1]],"date-time":"2025-11-01T01:45:16Z","timestamp":1761961516000},"source":"Crossref","is-referenced-by-count":0,"title":["Rationally designed minimized TbpB confers broad protection against meningococcal infection"],"prefix":"10.64898","author":[{"given":"Epshita A.","family":"Islam","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamie E.","family":"Fegan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory B.","family":"Cole","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles","family":"Calmettes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dixon","family":"Ng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalie Y. T.","family":"Au","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolyn M.","family":"Buckwalter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sang K.","family":"Ahn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2749-869X","authenticated-orcid":false,"given":"David M.","family":"Curran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura-lee","family":"Caruso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony B.","family":"Schryvers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trevor F.","family":"Moraes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1477-3616","authenticated-orcid":false,"given":"Scott D.","family":"Gray-Owen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"2","key":"2025110400500530000_2025.10.30.685603v1.1","first-page":"1","article-title":"Patterns of structural and sequence variation within isotype lineages of the Neisseria meningitidis transferrin receptor system","volume":"4","year":"2015","journal-title":"Microbiology Open"},{"key":"2025110400500530000_2025.10.30.685603v1.2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"2025110400500530000_2025.10.30.685603v1.3","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.7.4281-4287.2005"},{"issue":"1","key":"2025110400500530000_2025.10.30.685603v1.4","doi-asserted-by":"crossref","first-page":"10377","DOI":"10.1038\/s41598-017-10627-0","article-title":"New insights about functional and cross-reactive properties of antibodies generated against recombinant TbpBs of Haemophilus parasuis","volume":"7","year":"2017","journal-title":"Sci Rep"},{"key":"2025110400500530000_2025.10.30.685603v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2005.01.051"},{"issue":"10","key":"2025110400500530000_2025.10.30.685603v1.6","doi-asserted-by":"crossref","first-page":"1590","DOI":"10.1093\/infdis\/jix162","article-title":"Discordant Effects of Licensed Meningococcal Serogroup B Vaccination on Invasive Disease and Nasal Colonization in a Humanized Mouse Model","volume":"215","year":"2017","journal-title":"J Infect Dis"},{"key":"2025110400500530000_2025.10.30.685603v1.7","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444913022750"},{"issue":"3","key":"2025110400500530000_2025.10.30.685603v1.8","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1038\/nsmb.2251","article-title":"The structural basis of transferrin iron sequestration by transferrin binding protein B","volume":"19","year":"2012","journal-title":"Nature Structural and Molecular Biology"},{"key":"2025110400500530000_2025.10.30.685603v1.9","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1998.00710.x"},{"key":"2025110400500530000_2025.10.30.685603v1.10","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20052442"},{"key":"2025110400500530000_2025.10.30.685603v1.11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002027"},{"key":"2025110400500530000_2025.10.30.685603v1.12","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"2025110400500530000_2025.10.30.685603v1.13","doi-asserted-by":"crossref","unstructured":"Fegan, J.E. , Calmettes, C. , Islam, E.A. , Ahn, S.K. , Chaudhuri, S. , Yu, R.H. , et al. (2019). Utility of Hybrid Transferrin Binding Protein Antigens for Protection Against Pathogenic Neisseria Species. Frontiers in Immunology 10(247), eCollection 2019.","DOI":"10.3389\/fimmu.2019.00247"},{"issue":"1","key":"2025110400500530000_2025.10.30.685603v1.14","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1038\/s41541-024-01054-0","article-title":"Rational selection of TbpB variants yields a bivalent vaccine with broad coverage against Neisseria gonorrhoeae","volume":"10","year":"2025","journal-title":"NPJ Vaccines"},{"key":"2025110400500530000_2025.10.30.685603v1.15","doi-asserted-by":"crossref","first-page":"113037","DOI":"10.1016\/j.jim.2021.113037","article-title":"Development of a non-biased, high-throughput ELISA for the rapid evaluation of immunogenicity and cross-reactivity","volume":"493","year":"2021","journal-title":"Journal of Immunological Methods"},{"key":"2025110400500530000_2025.10.30.685603v1.16","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1222267"},{"key":"2025110400500530000_2025.10.30.685603v1.17","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.02572-14"},{"key":"2025110400500530000_2025.10.30.685603v1.18","doi-asserted-by":"publisher","DOI":"10.1080\/00325481.2016.1203238"},{"key":"2025110400500530000_2025.10.30.685603v1.19","doi-asserted-by":"crossref","unstructured":"Hooda, Y. , Shin, H.E. , Bateman, T.J. , and Moraes, T.F. (2017). Neisserial surface lipoproteins: structure, function and biogenesis. Pathog Dis 75(2).","DOI":"10.1093\/femspd\/ftx010"},{"key":"2025110400500530000_2025.10.30.685603v1.20","doi-asserted-by":"crossref","first-page":"126709","DOI":"10.1016\/j.vaccine.2025.126709","article-title":"Adjuvant-dependent impacts on vaccine-induced humoral responses and protection in preclinical models of nasal and genital colonization by pathogenic Neisseria","volume":"48","year":"2025","journal-title":"Vaccine"},{"key":"2025110400500530000_2025.10.30.685603v1.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003509"},{"key":"2025110400500530000_2025.10.30.685603v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.3267"},{"key":"2025110400500530000_2025.10.30.685603v1.23","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2010.07289.x"},{"issue":"3","key":"2025110400500530000_2025.10.30.685603v1.24","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1128\/iai.63.3.884-890.1995","article-title":"Evaluation of transferrin-binding protein 2 within the transferrin-binding protein complex as a potential antigen for future meningococcal vaccines","volume":"63","year":"1995","journal-title":"Infection and Immunity"},{"key":"2025110400500530000_2025.10.30.685603v1.25","doi-asserted-by":"publisher","DOI":"10.1111\/febs.14696"},{"key":"2025110400500530000_2025.10.30.685603v1.26","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMa1900236"},{"issue":"3","key":"2025110400500530000_2025.10.30.685603v1.27","doi-asserted-by":"crossref","first-page":"e609","DOI":"10.1093\/cid\/ciaa1733","article-title":"Effectiveness of Meningococcal Vaccines at Reducing Invasive Meningococcal Disease and Pharyngeal Neisseria meningitidis Carriage: A Systematic Review and Meta-analysis","volume":"73","year":"2021","journal-title":"Clin Infect Dis"},{"issue":"1","key":"2025110400500530000_2025.10.30.685603v1.28","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1080\/14760584.2022.2003708","article-title":"4CMenB vaccine and its role in preventing transmission and inducing herd immunity","volume":"21","year":"2022","journal-title":"Expert Rev Vaccines"},{"key":"2025110400500530000_2025.10.30.685603v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2009.06.029"},{"key":"2025110400500530000_2025.10.30.685603v1.30","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00020-08"},{"key":"2025110400500530000_2025.10.30.685603v1.31","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(97)76066-X"},{"key":"2025110400500530000_2025.10.30.685603v1.32","doi-asserted-by":"publisher","DOI":"10.1080\/10409238.2017.1293606"},{"key":"2025110400500530000_2025.10.30.685603v1.33","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.7.3945-3953.2005"},{"key":"2025110400500530000_2025.10.30.685603v1.34","doi-asserted-by":"crossref","unstructured":"Qamsari, M.M. , Rasooli, I. , Chaudhuri, S. , Astaneh, S.D.A. , and Schryvers, A.B. (2020). Hybrid Antigens Expressing Surface Loops of ZnuD From Acinetobacter baumannii Is Capable of Inducing Protection Against Infection. Front Immunol 11(158).","DOI":"10.3389\/fimmu.2020.00158"},{"key":"2025110400500530000_2025.10.30.685603v1.35","doi-asserted-by":"publisher","DOI":"10.1006\/mpat.1998.0226"},{"issue":"1","key":"2025110400500530000_2025.10.30.685603v1.36","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1128\/iai.65.1.55-63.1997","article-title":"Evaluation of recombinant transferrin binding protein B variants from Neisseria meningitidis for their ability of induce cross reactive and bactericidal antibodies against a genetically diverse collection of serogroup B strains","volume":"65","year":"1997","journal-title":"Infection and Immunity"},{"key":"2025110400500530000_2025.10.30.685603v1.37","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.9.4938-4947.2000"},{"key":"2025110400500530000_2025.10.30.685603v1.38","doi-asserted-by":"publisher","DOI":"10.1586\/14760584.2015.1016915"},{"key":"2025110400500530000_2025.10.30.685603v1.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2012.01.033"},{"key":"2025110400500530000_2025.10.30.685603v1.40","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199807233390401"},{"key":"2025110400500530000_2025.10.30.685603v1.41","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-691-7_2"},{"key":"2025110400500530000_2025.10.30.685603v1.42","doi-asserted-by":"crossref","first-page":"2405","DOI":"10.3389\/fimmu.2018.02405","article-title":"Intranasal Vaccination With Lipoproteins Confers Protection Against Pneumococcal Colonisation","volume":"9","year":"2018","journal-title":"Frontiers in Immunology"},{"key":"2025110400500530000_2025.10.30.685603v1.43","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4582"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.10.30.685603","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:56:04Z","timestamp":1768470964000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.10.30.685603"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,31]]},"references-count":43,"URL":"https:\/\/doi.org\/10.1101\/2025.10.30.685603","relation":{},"subject":[],"published":{"date-parts":[[2025,10,31]]},"subtype":"preprint"}}