Faculty of Pharmacy, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Comenius University Bratislava, Bratislava, Slovakia.
Faculty of Pharmacy, Toxicological and Antidoping Center, Comenius University Bratislava, Bratislava, Slovakia.
J Sep Sci. 2023 Aug;46(15):e2300244. doi: 10.1002/jssc.202300244. Epub 2023 May 26.
Capillary electrophoresis is recognized as a valued separation technique for its high separation efficiency, low sample consumption, good economic and ecological aspects, reproducibility, and complementarity to traditional liquid chromatography techniques. Capillary electrophoresis experiments are generally performed utilizing optical detection, such as ultraviolet or fluorescence detectors. However, in order to provide structural information, capillary electrophoresis hyphenated to highly sensitive and selective mass spectrometry has been developed to overcome the limitations of optical detections. Capillary electrophoresis-mass spectrometry is increasingly popular in protein analysis, including biopharmaceutical and biomedical research. It is frequently applied for the determination of physicochemical and biochemical parameters of proteins, offers excellent performance for in-depth characterizations of biopharmaceuticals at various levels of analysis, and has been also already proven as a promising tool in biomarker discovery. In this review, we focus on the possibilities and limitations of capillary electrophoresis-mass spectrometry for protein analysis at their intact level. Various capillary electrophoresis modes and capillary electrophoresis-mass spectrometry interfaces, as well as approaches to prevent protein adsorption and to enhance sample loading capacity, are discussed and the recent (2018-March 2023) developments and applications in the field of biopharmaceutical and biomedical analysis are summarized.
毛细管电泳因其高分离效率、低样品消耗、良好的经济和生态方面、重现性以及对传统液相色谱技术的互补性，被公认为一种有价值的分离技术。毛细管电泳实验通常使用光学检测，如紫外或荧光检测器进行。然而，为了提供结构信息，已经开发出与高灵敏度和选择性质谱联用的毛细管电泳技术，以克服光学检测的局限性。毛细管电泳-质谱联用技术在蛋白质分析中越来越受欢迎，包括生物制药和生物医学研究。它常用于测定蛋白质的物理化学和生化参数，在各种分析水平上对生物制药进行深入表征具有优异的性能，并且已经被证明是一种有前途的生物标志物发现工具。在这篇综述中，我们专注于毛细管电泳-质谱联用技术在蛋白质整体分析中的可能性和局限性。讨论了各种毛细管电泳模式和毛细管电泳-质谱接口，以及防止蛋白质吸附和提高样品加载能力的方法，并总结了该领域在生物制药和生物医学分析中的最新（2018 年 3 月至 2023 年）发展和应用。
