{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T06:05:52Z","timestamp":1780380352484,"version":"3.54.1"},"reference-count":34,"publisher":"Wiley","issue":"13","license":[{"start":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T00:00:00Z","timestamp":1776211200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T00:00:00Z","timestamp":1776211200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019124","name":"Asia University Hospital","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100019124","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2026,7,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Rationale<\/jats:title>\n                    <jats:p>Microwave\u2010accelerated proteolysis with optimized buffers reduces artificial N\/Q deamidation introduced during conventional peptide mapping of monoclonal antibodies. This workflow preserves CQA accuracy while significantly reducing digestion time and improving analytical throughput, offering a reliable, high\u2010efficiency strategy for biopharmaceutical quality control and deamidation assessment in therapeutic protein characterization.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Synthetic PENNY peptides, a murine mAb peptide (VNSAAFPAPIEK), and human IgG were reduced, alkylated, and trypsin\u2010digested in Tris or ammonium bicarbonate using conventional or microwave\u2010assisted protocols (900\u2009W, 1\u2009min; 100\u2009W, 30\u2009min). Deamidation was quantified by UPLC\u2010ESI\u2010MRM on a triple quadrupole MS. Peptide mapping and PTM localization were achieved by LC\u2010ESI\u2010MS\/MS, respectively, using a high\u2010resolution SCIEX TripleTOF 6600 and a low\u2010resolution Thermo LTQ XL with CID, ETD, and CID\u2009+\u2009ETD.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Buffer chemistry and exposure time strongly influenced artificial deamidation. Compared with overnight digestion (16\u201318\u2009h), microwave\u2010assisted processing reduced deamidation (4.0%\u20134.2% vs. 16.9%) while shortening preparation time. Optimized Tris conditions lowered PENNY peptide deamidation by ~60%\u201380%. For VNSAAFPAPIEK, high\u2010power microwaving reduced processing to 11\u2009min while preserving fragmentation coverage. Application to IgG decreased multisite N\/Q deamidation, and BSA validation confirmed consistently low deamidation (&lt;\u20091%), supporting robustness across sequence\u2010diverse proteins.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Microwave\u2010accelerated proteolysis under optimized Tris buffer conditions minimizes artificial N\/Q deamidation during peptide mapping while substantially reducing preparation time. Digestion time decreases from 16\u201318\u2009h to minutes, lowering deamidation by ~60%\u201380%. Consistent findings in IgG and BSA improve confidence in CQA assessment and support high\u2010throughput biopharmaceutical quality control.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/rcm.70081","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T09:29:18Z","timestamp":1776245358000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Microwave\u2010Accelerated Proteolysis Minimizes Artificial Deamidation and Enhances Speed for Peptide Mapping of Protein Pharmaceuticals"],"prefix":"10.1002","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-6925-1040","authenticated-orcid":false,"given":"Yi\u2010Feng","family":"Zheng","sequence":"first","affiliation":[{"name":"Institute of Molecular Biology National Chung Hsing University  Taichung 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