Aptamers, as novel recognition molecules, hold immense potential across various domains such as biosensing, nucleic acid drugs, medical diagnostics, as well as environmental and food analysis. The majority of aptamer selection processes targeting small molecules and protein commonly employ magnetic bead-based methodologies, wherein the target is initially immobilized on magnetic beads, followed by magnetic separation. The Evolutionary Systematic Evolution of Ligands by Exponential Enrichment technique based on capillary electrophoresis (CE-SELEX) is acknowledged as one of the most efficient screening methods. Our research group has achieved breakthroughs in employing CE-SELEX for the selection of aptamers targeting small molecules. This paper outlines specific methodologies utilized from 2005 to 2023 for CE-SELEX screening for small-molecule targets. It summarizes the methods for the separation of small molecules and oligonucleotide complexes, as well as the identification of candidate aptamers. Drawing upon our research group's extensive experience in CE-SELEX for selecting aptamers targeting multi-scale targets, we offer strategic guidance specifically tailored to the screening of aptamers for small-molecule targets using CE-SELEX. This includes systematic insights into each technical aspect of the screening process: analysis of the structure of small-molecule targets and characteristics of ssDNA libraries, patterns of CE separation and collection of complexes, screening strategies, and CE-based methods for the affinity and specificity characterization of aptamers. This comprehensive review aims to contribute to the widespread adoption of CE-SELEX technology, enhancing the efficiency and success rate of selecting aptamers for small-molecule targets.

中文翻译：

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毛细管电泳靶向小分子的适配子的选择：进展、挑战与展望


适配子作为新型识别分子，在生物传感、核酸药物、医学诊断以及环境和食品分析等各个领域都具有巨大的潜力。大多数靶向小分子和蛋白质的适配体选择过程通常采用基于磁珠的方法，其中靶标最初固定在磁珠上，然后进行磁性分离。基于毛细管电泳的指数富集技术 （CE-SELEX） 的配体进化系统进化被认为是最有效的筛选方法之一。我们的研究小组在使用 CE-SELEX 选择靶向小分子的适配子方面取得了突破。本文概述了 2005 年至 2023 年用于小分子靶标 CE-SELEX 筛选的具体方法。它总结了小分子和寡核苷酸复合物的分离方法，以及候选适配子的鉴定方法。利用我们研究小组在 CE-SELEX 选择靶向多尺度靶点的适配子方面的丰富经验，我们提供专门针对使用 CE-SELEX 筛选小分子靶点的适配子的战略指导。这包括对筛选过程每个技术方面的系统见解：小分子靶标结构和 ssDNA 文库特性分析、CE 分离和复合物收集模式、筛选策略以及基于 CE 的适配子亲和力和特异性表征方法。本综述旨在为 CE-SELEX 技术的广泛采用做出贡献，提高为小分子靶标选择适配子的效率和成功率。