Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, the Netherlands.
Biomedical Microscale Analytics, Leiden Academic Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, the Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jun 15;1118-1119:116-136. doi: 10.1016/j.jchromb.2019.04.020. Epub 2019 Apr 12.
Capillary electrophoresis (CE) played an important role in developments in the life sciences. The technique is nowadays used for the analysis of both large and small molecules in applications where it performs better than or is complementary to liquid chromatographic techniques. In this review, principles of different electromigration techniques, especially capillary isoelectric focusing (CIEF), capillary gel (CGE) and capillary zone electrophoresis (CZE), are described and recent developments in instrumentation, with an emphasis on mass spectrometry (MS) coupling and microchip CE, are discussed. The role of CE in the life sciences is shown with applications in which it had a high impact over the past few decades. In this context, current practice for the characterization of biopharmaceuticals (therapeutic proteins) is shown with CIEF, CGE and CZE using different detection techniques, including MS. Subsequently, the application of CGE and CZE, in combination with laser induced fluorescence detection and CZE-MS are demonstrated for the analysis of protein-released glycans in the characterization of biopharmaceuticals and glycan biomarker discovery in biological samples. Special attention is paid to developments in capillary coatings and derivatization strategies for glycans. Finally, routine CE analysis in clinical chemistry and latest developments in metabolomics approaches for the profiling of small molecules in biological samples are discussed. The large number of CE applications published for these topics in recent years clearly demonstrates the established role of CE in life sciences.
毛细管电泳 (CE) 在生命科学的发展中发挥了重要作用。如今,该技术用于分析大分子和小分子,在某些应用中,其性能优于或互补于液相色谱技术。在这篇综述中,描述了不同电泳技术的原理,特别是毛细管等电聚焦 (CIEF)、毛细管凝胶电泳 (CGE) 和毛细管区带电泳 (CZE),并讨论了仪器的最新进展,重点是质谱 (MS) 联用和微流控芯片 CE。通过展示 CE 在过去几十年中具有重大影响的应用,说明了它在生命科学中的作用。在这方面,展示了 CIEF、CGE 和 CZE 用于生物制药(治疗蛋白)特性分析的当前实践,包括使用不同的检测技术,包括 MS。随后,演示了 CGE 和 CZE 与激光诱导荧光检测和 CZE-MS 的联用,用于分析生物制药中蛋白质释放的聚糖以及生物样品中聚糖生物标志物的发现。特别关注了用于聚糖衍生化的毛细管涂层和衍生化策略的发展。最后,讨论了临床化学中的常规 CE 分析以及生物样品中小分子分析的代谢组学方法的最新进展。近年来,针对这些主题发表的大量 CE 应用清楚地表明了 CE 在生命科学中的既定作用。