{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,11,7]],"date-time":"2026-11-07T04:16:27Z","timestamp":1794024987283,"version":"4.8.0"},"posted":{"date-parts":[[2026,5,16]]},"group-title":"Biochemistry","reference-count":70,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T00:00:00Z","timestamp":1778889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH","award":["P41 GM108538"],"award-info":[{"award-number":["P41 GM108538"]}]},{"name":"NIH","award":["R35 GM118110"],"award-info":[{"award-number":["R35 GM118110"]}]},{"name":"NIH","award":["R35 GM126914"],"award-info":[{"award-number":["R35 GM126914"]}]},{"id":[{"id":"https:\/\/ror.org\/01y2jtd41","id-type":"ROR","asserted-by":"publisher"}]},{"id":[{"id":"https:\/\/ror.org\/05cb4rb43","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Peptide mapping is a critical tool for characterizing biotherapeutic proteins and is essential for the development of monoclonal antibody drugs. Here we describe a new direct infusion technology that streamlines peptide mapping data collection and analysis, accelerating the method by up to 100-fold. This method, which we term RaPiD-mAb-MS, combines high-throughput plate-based sample preparation with direct infusion mass spectrometry analysis. RaPiD-mAb-MS allows analysis of 96 samples within \u223c 1.5 to 2 hours, routinely achieves &gt;95% sequence coverage, and has been successfully applied to 28 unique antibodies and over 2,000 samples. Here we demonstrate that RaPiD-mAb-MS detects and quantifies oxidation, deamidation, isomerization, glycosylation, and sequence variants with results comparable to conventional LC-MS based methods in a fraction of the time. Further, by eliminating chromatography, data analysis is greatly streamlined and simplified. By allowing for the collection of \u223c 1,000 peptide maps per day, RaPiD-mAb-MS is positioned to accelerate all phases of antibody-based drug discovery &amp; development and sets the stage for collection of massive datasets that would allow artificial intelligent prediction of optimal antibody variants and formulations.<\/jats:p>","DOI":"10.64898\/2026.05.14.725248","type":"posted-content","created":{"date-parts":[[2026,5,17]],"date-time":"2026-05-17T04:35:14Z","timestamp":1778992514000},"source":"Crossref","is-referenced-by-count":1,"title":["Rapid Peptide Mapping of Monoclonal Antibodies with Direct Infusion Mass Spectrometry"],"prefix":"10.64898","author":[{"given":"Austin Z.","family":"Salome","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"},{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel","family":"Morgenstern","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander S.","family":"Hebert","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Craig D.","family":"Wenger","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2653-1702","authenticated-orcid":false,"given":"Pavel","family":"Sinitcyn","sequence":"additional","affiliation":[{"name":"Morgridge Institute for Research, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benton J.","family":"Anderson","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John S.","family":"Chlystek","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lia R.","family":"Serrano","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"},{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keaton L.","family":"Mertz","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"},{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian J.","family":"Miller","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erik","family":"Miller-Galow","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Chemistry, University of Wisconsin\u2013Madison, Madison, WI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malika P.","family":"Godamudunage","sequence":"additional","affiliation":[{"name":"AbbVie, Inc. 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