{"title":"Gel electrophoresis-based proteoform separation and analysis.","authors":"Paul Dowling, Kay Ohlendieck","doi":"10.1016/bs.acc.2025.04.002","DOIUrl":null,"url":null,"abstract":"<p><p>Proteomics using gel electrophoresis (GE) for efficient protein separation prior to mass spectrometry is a frequently employed and proteoform-centric analysis tool of biomedical chemistry. Isolated molecular species can be visualized by various protein staining techniques and their characterization being combined with protein interaction analyses and the determination of dynamic post-translational modifications. This chapter describes the bioanalytical usefulness of one-dimensional GE (1D-GE) approaches, including the gel electrophoresis liquid chromatography (GeLC) mass spectrometry method, as well as commonly used two-dimensional gel electrophoresis (2D-GE) techniques, including fluorescence difference GE (DIGE) for sophisticated comparative studies. The ways in which advanced protein identification and gel-based proteomics can help to discover novel biomarker candidates is discussed.</p>","PeriodicalId":101297,"journal":{"name":"Advances in clinical chemistry","volume":"127 ","pages":"119-171"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advances in clinical chemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/bs.acc.2025.04.002","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/5/15 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Proteomics using gel electrophoresis (GE) for efficient protein separation prior to mass spectrometry is a frequently employed and proteoform-centric analysis tool of biomedical chemistry. Isolated molecular species can be visualized by various protein staining techniques and their characterization being combined with protein interaction analyses and the determination of dynamic post-translational modifications. This chapter describes the bioanalytical usefulness of one-dimensional GE (1D-GE) approaches, including the gel electrophoresis liquid chromatography (GeLC) mass spectrometry method, as well as commonly used two-dimensional gel electrophoresis (2D-GE) techniques, including fluorescence difference GE (DIGE) for sophisticated comparative studies. The ways in which advanced protein identification and gel-based proteomics can help to discover novel biomarker candidates is discussed.