{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:19:23Z","timestamp":1792790363385,"version":"4.5.0"},"posted":{"date-parts":[[2026,9,16]]},"group-title":"Infectious Diseases (except HIV\/AIDS)","reference-count":83,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"name":"National Institute of Allergy and Infectious Diseases","id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,16]]},"abstract":"<jats:title>\n                  <jats:bold>Abstract<\/jats:bold>\n                <\/jats:title>\n                <jats:p>Sequential dengue virus (DENV) infection is thought to induce long-lived protection against dengue. However, little is known about the specificity, phenotype, and repertoire of the enduring B cell response due to limitations in high-throughput methods. We used stabilized DENV envelope (E) dimers as probes for in-depth memory B cell repertoire characterization using two complementary sequencing strategies. Using Rapid Assembly, Transfection, and Production of Immunoglobulins (RATP-Ig), we isolated 227 DENV2 E dimer-specific memory B cells from an immune donor, of which 87% bound at least two serotypes, 68% bound the fusion loop (FL-sensitive), and 42% were neutralizing. Among the most neutralizing, cross-binding antibodies, the FL-sensitive antibodies had weaker but broad neutralization, including to other flaviviruses, whereas FL-\u2018agnostic\u2019 antibodies mapped to E domain II and III and were potently neutralizing, but only against one or two serotypes. Competitive binding of the DENV1-4 E dimers distinguished the more potently neutralizing FL-agnostic antibodies and broad and potent anti (\u03b1)- envelope dimer epitope (EDE) antibodies from the low-quality FL-sensitive antibodies. Further, analysis of the heavy chain complementarity determining region 3 (CDRH3) showed FL-sensitive and \u03b1-EDE antibodies were more negatively charged, but \u03b1-EDE antibodies were distinguished by longer and more aromatic CDRH3s. We then demonstrated how oligonucleotide-tagged DENV1-4 dimers and proteogenomic single-cell B cell receptor sequencing (LIBRA-seq) could identify memory B cells predicted to have potent neutralization based on DENV-binding and CDRH3 physiochemical properties. This study provides insight into the secondary DENV-specific B cell repertoire and approaches for identifying protective antibodies as part of large-screening efforts.<\/jats:p>\n                <jats:sec>\n                  <jats:title>One Sentence Summary<\/jats:title>\n                  <jats:p>Approaches to define the genetic, physicochemical, and functional properties of dengue virus envelope dimer-specific memory B cells after secondary infection.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.15.26363039","type":"posted-content","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T01:57:12Z","timestamp":1789610232000},"source":"Crossref","is-referenced-by-count":0,"title":["Signatures of the neutralizing dengue virus-reactive memory B cell repertoire identified by multiplexed envelope dimer probes"],"prefix":"10.64898","author":[{"given":"Kelsey E.","family":"Lowman","sequence":"first","affiliation":[{"name":"Viral Epidemiology and Immunology Unit, 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