Kawai Takayuki
Department of Chemistry, Faculty of Science, Kyushu University.
Nihon Yakurigaku Zasshi. 2024;159(5):321-326. doi: 10.1254/fpj.24036.
In recent years, various trace bioanalysis methods have been developed, including single-cell transcriptome analysis methods. As the sample volume and amount of biomolecules contained therein are extremely limited, development of new single-cell analysis methods require extremely high-level techniques. It is necessary to design an appropriate analysis system that integrates a highly sensitive detection system and a pretreatment protocol for minimizing sample loss, where separation method is especially important for analyzing diverse mixtures of biomolecules. Among them, capillary electrophoresis (CE) can separate biomolecules in nanoliter-scale solutions with high resolution, making it highly compatible with trace samples such as single cells. By combining with highly sensitive nano-electrospray ionization-mass spectrometry (MS), it is possible to detect nanomolar to sub-nanomolar biomolecules, which can be further improved by using online sample preconcentration methods. These highly sensitive analytical techniques have made it possible to analyze trace amounts of metabolites, proteins, lipids, etc. This review paper summarizes the research on CE-MS trace bioanalysis that has been reported to date, with a focus on single-cell analysis.
近年来，已经开发了各种痕量生物分析方法，包括单细胞转录组分析方法。由于样品体积及其所含生物分子的量极其有限，新的单细胞分析方法的开发需要极高水平的技术。有必要设计一个合适的分析系统，该系统集成了高灵敏度检测系统和用于最小化样品损失的预处理方案，其中分离方法对于分析生物分子的各种混合物尤为重要。其中，毛细管电泳（CE）可以在纳升规模的溶液中高分辨率地分离生物分子，使其与单细胞等痕量样品高度兼容。通过与高灵敏度的纳米电喷雾电离质谱（MS）相结合，可以检测纳摩尔至亚纳摩尔的生物分子，通过使用在线样品预浓缩方法可以进一步提高检测能力。这些高灵敏度分析技术使得分析痕量的代谢物、蛋白质、脂质等成为可能。本文综述了迄今为止报道的关于CE-MS痕量生物分析的研究，重点是单细胞分析。
