{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T16:14:59Z","timestamp":1787242499136,"version":"build-2736575974"},"reference-count":55,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2023YFF0614603"],"award-info":[{"award-number":["2023YFF0614603"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22374009"],"award-info":[{"award-number":["22374009"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["advanced.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Advanced Science"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>The sequencing of synthetic oligonucleotides is fundamental to life sciences and therapeutics, yet current analytical approaches present inherent technical drawbacks of varying degrees. Hybridization\u2010based techniques cannot sequence the oligonucleotide primers they rely on, while the accuracy of mass spectrometry (MS)\u2010based techniques significantly diminishes as fragment length increases. Here, we introduce a liquid\u2010phase ion trap (LPIT) coupled with electrospray ionization (ESI)\u2010MS as a solution for rapid, high\u2010fidelity oligonucleotide sequencing. The LPIT utilizes a strong electric field to efficiently dissociate confounding metal cation adducts prior to MS analysis, resulting in cleaner spectra and enhanced signal intensity. This \u201cdesalting\u2010first\u201d approach enables direct, rapid oligonucleotide analysis, achieving mass measurement accuracy comparable to benchmark liquid chromatography\u2010ESI\u2010MS (LC\u2010ESI\u2010MS) platforms. By integrating this platform with an exonuclease digestion workflow, we achieved full sequence coverage of oligonucleotides in under 30 min, a dramatic improvement in speed over existing methods. This work establishes LPIT\u2010ESI\u2010MS as a powerful platform for the direct and rapid sequencing of long oligonucleotides, addressing a critical unmet need in biotechnology and drug development.<\/jats:p>","DOI":"10.1002\/advs.77313","type":"journal-article","created":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T10:03:20Z","timestamp":1787220200000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid Exonuclease\u2010Based\n                    <i>De Novo<\/i>\n                    Sequencing Strategy for Long Oligonucleotides via Liquid\u2010Phase Ion Trap Coupled With Mass Spectrometry"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-9378-5777","authenticated-orcid":false,"given":"Song","family":"Lu","sequence":"first","affiliation":[{"name":"Center for Advanced Measurement Science National Institute of Metrology  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Hong","sequence":"additional","affiliation":[{"name":"Kunshan Nier Precision Instrumentation Inc.  Kunshan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyan","family":"Fu","sequence":"additional","affiliation":[{"name":"School of Medical Technology Beijing Institute of Technology  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2887-4995","authenticated-orcid":false,"given":"Di","family":"Wang","sequence":"additional","affiliation":[{"name":"Center for Advanced Measurement Science National Institute of Metrology  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunhua","family":"Gao","sequence":"additional","affiliation":[{"name":"Center for Advanced Measurement Science National Institute of Metrology  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5274-1546","authenticated-orcid":false,"given":"Wei","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Medical Technology Beijing 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