Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
Electrophoresis. 2019 Mar;40(5):625-642. doi: 10.1002/elps.201800367. Epub 2019 Jan 11.
Nearly all processes in living organisms are controlled and regulated by the synergy of many biomolecule interactions involving proteins, peptides, nucleic acids, nucleotides, saccharides, and small molecular weight ligands. There is growing interest in understanding them, not only for the purposes of interactomics as an essential part of system biology, but also in their further elucidation in disease pathology, diagnostics, and treatment. The necessity of detailed investigation of these interactions leads to the requirement of laboratory methods characterized by high efficiency and sensitivity. As a result, many instrumental approaches differing in their fundamental principles have been developed, including those based on capillary electrophoresis. Although capillary electrophoresis offers numerous advantages for such studies, it still has one serious limitation, its poor concentration sensitivity with the most commonly used detection method-ultraviolet-visible spectrometry. However, coupling capillary electrophoresis with a more sensitive detector fulfils the above-mentioned requirement. In this review, capillary electrophoresis combined with fluorescence, mass spectrometry, and several nontraditional detection techniques in affinity interaction studies are summarized and discussed, together with the possibility of conducting these measurements in microchip format.
几乎所有生物体的过程都是通过涉及蛋白质、肽、核酸、核苷酸、糖和小分子配体的许多生物分子相互作用的协同作用来控制和调节的。人们越来越感兴趣地了解这些相互作用，不仅是为了作为系统生物学的一个重要组成部分的相互作用组学，而且还为了进一步阐明它们在疾病病理学、诊断和治疗中的作用。对这些相互作用进行详细研究的必要性导致需要具有高效率和灵敏度的实验室方法。因此，已经开发了许多基于不同基本原理的仪器方法，包括基于毛细管电泳的方法。尽管毛细管电泳在这些研究中具有许多优点，但它仍然存在一个严重的限制，即其最常用的检测方法-紫外-可见光谱法的浓度灵敏度较差。然而，将毛细管电泳与更灵敏的检测器结合使用可以满足上述要求。在本文中，总结和讨论了毛细管电泳与荧光、质谱和几种非传统检测技术在亲和相互作用研究中的结合应用，以及在微芯片格式中进行这些测量的可能性。
