{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T20:23:16Z","timestamp":1785874996534,"version":"3.56.0"},"reference-count":17,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    This study presents a capillary gel electrophoresis\u2013laser\u2010induced fluorescence (CGE\u2013LIF) method for analyzing topoisomers of large plasmid DNA (up to 19\u00a0kbp) using an uncoated capillary. Separation of large plasmid isoforms, particularly the open circular (OC) form, poses significant challenges in gel buffer formulation. To address this, we systematically investigated the effects of multiple parameters\u2014including polymer type, fluorescent dye selection, stir bar geometry, stirring speed and duration, gel volume, container dimensions, and technique for polymer addition\u2014on separation performance. The optimized gel buffer composition was 0.9% polyethylene oxide (PEO), 0.02% SYBR Green, 50\u00a0mM\n                    <jats:italic>N<\/jats:italic>\n                    \u2010(2\u2010acetamido)\u20102\u2010aminoethanesulfonic acid (ACES), 1 mM EDTA, and 20\u00a0mM NaCl at pH 6.25. Replacing hydroxypropyl methyl cellulose (HPMC) gel with PEO gel significantly improved OC isoform separation. Furthermore, substituting SYBR Gold with SYBR Green enhanced the robustness of OC separation and yielded a 4\u2010 to 8\u2010fold increase in sensitivity. The PEO gel preparation process was successfully scaled and optimized at the 100\u00a0mL level. With consistent capillary cleaning and storage protocols, the method demonstrated excellent reproducibility over 150 consecutive runs.\n                  <\/jats:p>","DOI":"10.1002\/elps.70084","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T21:36:06Z","timestamp":1773005766000},"page":"320-327","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Sensitive Analysis of Topoisomers of Large Plasmid DNA by Capillary Gel Electrophoresis With Laser\u2010Induced Fluorescence in Uncoated Capillary"],"prefix":"10.1002","volume":"47","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8527-1982","authenticated-orcid":false,"given":"Yan","family":"He","sequence":"first","affiliation":[{"name":"Analytical Research and Development Pfizer Inc  Chesterfield Missouri USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Orlet","sequence":"additional","affiliation":[{"name":"Analytical Research and Development Pfizer Inc  Chesterfield Missouri USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivia","family":"Cunio","sequence":"additional","affiliation":[{"name":"Analytical Research and Development Pfizer Inc  Chesterfield Missouri USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas F","family":"Lerch","sequence":"additional","affiliation":[{"name":"Analytical Research and Development Pfizer Inc  Chesterfield Missouri USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan A.","family":"Lacher","sequence":"additional","affiliation":[{"name":"Analytical Research and Development Pfizer Inc  Chesterfield Missouri USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,3]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2012.11.006"},{"key":"e_1_2_8_3_1","volume-title":"The Production of Plasmid DNA Vaccine in Escherichia coli: A Novel Bacterial\u2010Based Vaccine Production Platform","author":"Chartrain M.","year":"2014"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2009.02.003"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)40095-1"},{"key":"e_1_2_8_6_1","first-page":"38","article-title":"Long\u2010Term Stability Study and Topology Analysis of Plasmid DNA by Capillary Gel Electrophoresis","volume":"4","author":"Schleef M.","year":"2006","journal-title":"Bioprocess International"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2697(03)00244-6"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.2174\/1566524020666200427110452"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202100343"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.201000061"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-021-03714-9"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.202300212"},{"key":"e_1_2_8_13_1","first-page":"1068","article-title":"Rapid Quantitative Analysis of Double\u2010Stranded Plasmid DNA With Capillary Gel Electrophoresis for Applications in Quality Control and Radiation Research","volume":"15","author":"Hahn M. B.","year":"2025","journal-title":"Nature Scientific Report Open Access"},{"key":"e_1_2_8_14_1","unstructured":"L.De MartininandR.Waser \u201cHigh\u2010Throughput Plasmid DNA Isoform Analysis Using Capillary Gel Electrophoresis \u201doral presentation at Cepharm 2025 Sep 8\u201310 Rockville MD."},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1039\/D3SM00687E"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-011-3890-1_20"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1998.3067"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.200600029"}],"container-title":["ELECTROPHORESIS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.70084","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/elps.70084","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.70084","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T10:40:06Z","timestamp":1776768006000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/elps.70084"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,3]]},"references-count":17,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026,4]]}},"alternative-id":["10.1002\/elps.70084"],"URL":"https:\/\/doi.org\/10.1002\/elps.70084","archive":["Portico"],"relation":{},"ISSN":["0173-0835","1522-2683"],"issn-type":[{"value":"0173-0835","type":"print"},{"value":"1522-2683","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,3]]},"assertion":[{"value":"2025-12-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-02-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-03","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}