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Capillaries are commonly coated with neutral or cationic polymers to reduce protein adsorption in CZE\u2010MS. We recently introduced a cationic polymer coating [poly(acrylamide\u2010co\u2010(3\u2010acrylamidopropyl) trimethylammonium chloride (PAMAPTAC)] to CZE\u2010MS\u2013based TDP. Here, we further improved the coating procedure to enhance the ease and repeatability of the cationic coating preparation by employing 2,2\u2032\u2010azobisisobutyronitrile (AIBN) as the radical initiator and methanol as the solvent. The new coating method not only simplifies the coating process but is also user\u2010friendly. Different laboratory members were able to reproduce this type of cationic coating with consistent CZE\u2010MS performance. We evaluated the performance of CZE\u2010MS with the cationic coating in analyzing various complex biological samples (nanoparticle protein corona, histone, and an\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    cell lysate) and assessed its long\u2010term reproducibility with an\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>E. coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    cell lysate for 35 runs (over 40\u2009h) of continuous measurement. The CZE\u2010MS system produced reproducible measurements for all the samples regarding separation profiles, proteoform migration time (MT), and proteoform intensity. For example, the system enabled the detection of large proteoforms (26\u201335\u2009kDa) from an\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>E. coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    sample with consistent MT, relative standard deviations (RSDs) &lt;7% without MT correction and 2.1% after MT alignment. We expect that the APTAC coating will further advance CZE\u2010MS\u2013based TDP for broad and large\u2010scale applications.\n                  <\/jats:p>","DOI":"10.1002\/jms.70005","type":"journal-article","created":{"date-parts":[[2025,12,27]],"date-time":"2025-12-27T03:03:51Z","timestamp":1766804631000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Reproducible Capillary Electrophoresis\u2013Mass Spectrometry\u2013Based Top\u2010Down Proteomics of Complex Proteomes Enabled by an Advanced Cationic Polymer Coating"],"prefix":"10.1002","volume":"61","author":[{"given":"Guangyao","family":"Gao","sequence":"first","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Fang","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yifan","family":"Yue","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex\u00a0T.","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qianjie","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qianyi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guijie","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8939-5042","authenticated-orcid":false,"given":"Liangliang","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Chemistry Michigan State University  East Lansing Michigan USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,26]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2222081120"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-018-0085-7"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.8b00693"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-025-01866-8"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M114.047480"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abq6348"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.202400336"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1200729"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.1c00893"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac503964j"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2023.2229102"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2015.06.003"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(01)95485-8"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(01)90205-5"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202200144"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.4c01119"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2024.343162"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1006\/mchj.1999.1718"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200305714"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2007.10.114"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac960432l"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202000204"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.201501349"},{"key":"e_1_2_8_25_1","unstructured":"M. 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