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Proteins were detected through their intrinsic fluorescence at 350\u00a0nm following excitation at 280\u00a0nm, enabling real\u2010time observation of migration behavior without additional labeling. Time\u2010resolved imaging allowed direct visualization of both migration and band broadening of mAbs under reducing and non\u2010reducing conditions throughout the separation process. Analysis of electrophoretic mobilities suggests that mAb separation in the gel matrix is largely governed by the Ogston sieving mechanism. Quantification of band broadening revealed that peak dispersion increased with applied electric field strength, as evidenced by the marked reduction in broadening when the separation voltage was removed. Qualitative evaluation of the dispersion behavior suggests that mAb band broadening arises from the combined effects of electromigration dispersion (EMD) and gel\u2010sieving\u2010induced dispersion, with glycosylation\u2010related heterogeneity providing an additional contribution for glycosylated species such as the heavy chain (HC) and intact mAb. These results demonstrate the utility of real\u2010time imaging for elucidating separation mechanisms and dispersion phenomena in CGE\u2013SDS and provide new insights into factors governing the electrophoretic behavior of therapeutic antibodies.<\/jats:p>","DOI":"10.1002\/elps.70131","type":"journal-article","created":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T12:36:32Z","timestamp":1783773392000},"page":"826-839","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Time\u2010Resolved Native Fluorescence Imaging of the Migration and Band Broadening of Monoclonal Antibody During Capillary Gel Electrophoresis With Sodium Dodecyl Sulfate"],"prefix":"10.1002","volume":"47","author":[{"given":"Hui","family":"Xu","sequence":"first","affiliation":[{"name":"R&amp;D Systems, a Bio\u2010Techne Brand  San Jose California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uyen","family":"Nguyen","sequence":"additional","affiliation":[{"name":"R&amp;D Systems, a Bio\u2010Techne Brand  San Jose California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom","family":"Yang","sequence":"additional","affiliation":[{"name":"R&amp;D Systems, a Bio\u2010Techne Brand  San Jose California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,11]]},"reference":[{"key":"e_1_2_9_2_1","volume-title":"Therapeutic Monoclonal Antibodies: From Bench to Clinic","author":"An Z.","year":"2011"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-61779-821-4_2"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1039\/C7AN00198C"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2019.04.020"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2024.116386"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202000199"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202100068"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1039\/tf9585401754"},{"key":"e_1_2_9_10_1","volume-title":"Protides of the Biological Fluids, 14th Colloquium","author":"Morris C. 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