色谱

色谱 ›› 2025, Vol. 43 ›› Issue (5): 424-433.DOI: 10.3724/SP.J.1123.2024.10029

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外泌体的核酸适配体筛选技术研究进展

郑李婷1, 杨歌2,*(), 屈锋1,*()   

  1. 1.北京理工大学生命学院, 北京 100081
    2.中国医学科学院北京协和医学院医药生物技术研究所, 北京 100050
  • 收稿日期:2024-10-26 出版日期:2025-05-08 发布日期:2025-05-07
  • 通讯作者: *Tel:(010)68918015,E-mail:qufengqu@bit.edu.cn(屈锋);Tel:(010)63125681,E-mail:yangge@imb.cams.cn(杨歌).
  • 基金资助:
    北京市自然科学基金(7222310);国家自然科学基金项目(82204263);国家自然科学基金项目(22274009);国家自然科学基金项目(9921874010)

Advances in exosome-targeting aptamer-screening techniques

ZHENG Liting1, YANG Ge2,*(), QU Feng1,*()   

  1. 1. School of Life Science, Beijing Institute of Technology, Beijing 100081, China
    2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
  • Received:2024-10-26 Online:2025-05-08 Published:2025-05-07
  • Supported by:
    Beijing Municipal Natural Science Foundation(7222310);National Natural Science Foundation of China(82204263);National Natural Science Foundation of China(22274009);National Natural Science Foundation of China(9921874010)

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摘要：

外泌体密切参与细胞间通讯,调控细胞生理过程,是目前癌症和其他疾病早期诊断的潜在生物标志物之一。因此,具有特定功能的外泌体的检测与分离具有重要临床意义。与此同时,开发低成本、高灵敏度的外泌体特异性识别元件,将在疾病早期诊断和治疗过程中发挥关键作用。核酸适配体是通过指数富集配体系统进化技术(systematic evolution of ligands by exponential enrichment, SELEX)筛选获得的、可与靶标特异性结合的单链脱氧核糖核酸或核糖核酸,具备稳定性高、可化学合成、高亲和力和特异性以及靶标范围广等独特优势。目前,靶向外泌体的适配体已应用于细胞成像、药物递送、疾病诊疗等众多研究领域。然而,外泌体的异质性以及复杂结构,为精准识别特定外泌体的适配体筛选带来巨大挑战,高效的筛选技术是获得优良性能适配体的关键。本文总结了外泌体适配体筛选过程中关键靶点的功能及其选择策略,概述了目前应用于外泌体适配体筛选的主要方法与应用,包括磁珠-SELEX、微流控-SELEX、硝酸纤维素膜-SELEX、细胞-SELEX和毛细管电泳-SELEX等5种筛选技术,并对其分离原理、优势与限制以及最新应用进行阐述与分析。最后,对外泌体适配体筛选所面临的问题与挑战进行了总结与展望。

关键词: 适配体, 指数富集系统进化技术, 外泌体

Abstract:

Exosomes play crucial intercellular-communication roles and regulate various cellular physiological processes. They are considered potential biomarkers for the early diagnosis of cancers and other diseases. Therefore, detecting and isolating exosomes with specific functions has significant clinical implications. Moreover, the development of low-cost, highly sensitive recognition elements for identifying exosomes is essential for advancing early disease diagnosis and treatment. Nucleic acid aptamers are single-stranded DNA or RNA molecules capable of specifically binding to targets and are produced through the systematic evolution of ligands by exponential enrichment (SELEX) technique. Such aptamers are highly stable, chemically synthesizable, exhibit high affinities and specificities, and are applicable to a broad range of targets, which endow them with unique advantages. Currently, aptamers that target exosomes have been used in a variety of research fields, including cell imaging, drug delivery, and disease diagnosis and treatment. However, selecting aptamers that precisely identify specific exosomes is significantly challenging owing to the complex structures of exosome and their heterogeneity. Consequently, obtaining high-performance aptamers requires efficient screening techniques. This review first summarizes the functions and selection strategies of key targets for exosome-aptamer screening. Furthermore, it outlines the main methods and techniques currently used to screen exosome aptamers, which includes five screening techniques: magnetic bead-SELEX, microfluidic-SELEX, nitrocellulose-SELEX, cell-SELEX, and capillary electrophoresis-SELEX. The separation principles, advantages, limitations, and the latest applications of these techniques are discussed in detail. The review finally addresses current challenges associated with selecting exosome aptamers and provides insight into future research directions.

Key words: aptamer, systematic evolution of ligands by exponential enrichment (SELEX), exosome

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