Hu Yangyang, Jiang Guangyu, Wen Yalun, Shao Yuchen, Yang Ge, Qu Feng
School of Life Science; Key Laboratory of Molecular Medicine and Biotherapy; Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering; Beijing Institute of Technology, Beijing 100081, China.
CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Antimicrobial Agents, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Biotechnol Adv. 2025 Jan-Feb;78:108491. doi: 10.1016/j.biotechadv.2024.108491. Epub 2024 Nov 26.
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.
适体作为新型识别分子，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潜力。大多数针对小分子和蛋白质的适体筛选过程通常采用基于磁珠的方法，即先将靶标固定在磁珠上，然后进行磁分离。基于毛细管电泳的指数富集配体系统进化技术（CE-SELEX）被认为是最有效的筛选方法之一。我们的研究小组在利用CE-SELEX筛选针对小分子的适体方面取得了突破。本文概述了2005年至2023年用于CE-SELEX筛选小分子靶标的具体方法。总结了小分子与寡核苷酸复合物的分离方法以及候选适体的鉴定方法。凭借我们研究小组在CE-SELEX筛选针对多尺度靶标的适体方面的丰富经验，我们提供了专门针对使用CE-SELEX筛选小分子靶标适体的策略指导。这包括对筛选过程各个技术环节的系统见解：小分子靶标结构分析和单链DNA文库特征、CE分离和复合物收集模式、筛选策略以及基于CE的适体亲和力和特异性表征方法。这篇综述旨在促进CE-SELEX技术的广泛应用，提高筛选小分子靶标适体的效率和成功率。
