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适配体筛选:多功能微流控平台及其多样应用

Aptamer selection versatile microfluidic platforms and their diverse applications.

作者信息

Chung Yi-Da, Tsai Yi-Cheng, Wang Chi-Hung, Lee Gwo-Bin

机构信息

Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.

Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan.

出版信息

Lab Chip. 2025 Feb 25;25(5):1047-1080. doi: 10.1039/d4lc00859f.

Abstract

Aptamers are synthetic oligonucleotides that bind with high affinity and specificity to various targets, making them invaluable for diagnostics, therapeutics, and biosensing. Microfluidic platforms can improve the efficiency and scalability of aptamer selection, especially through advancements in systematic evolution of ligands by exponential enrichment (SELEX) methods. Microfluidic SELEX methods are less time-consuming and labor-intensive and include critical steps like library preparation, binding, partitioning, and amplification. This review examines the contributions of microfluidic technology to SELEX-based aptamer identification, with alternative methods like conditional SELEX, -like SELEX and Non-SELEX for selecting aptamers and also discusses critical SELEX steps over the past decade. This work also examined the integrated microfluidic systems for SELEX, highlighting innovations such as conditional SELEX and -like SELEX. These advancements provide potential solutions to existing challenges in aptamer selection using conventional SELEX, especially concerning biological samples. A trend toward non-SELEX methods was also reviewed and discussed, wherein nucleic acid amplification was eliminated to improve aptamer selection. Microfluidic platforms have demonstrated versatility not only in aptamer selection but also in various detection applications; they allow for precise control of liquid flow and have been essential in the advancement of therapeutic aptamers, facilitating accurate screening, enhancing drug delivery systems, and enabling targeted therapeutic interventions. Although advances in microfluidic technology are expected to enhance aptamer-based diagnostics, therapeutics, and biosensing, challenges still persist, especially in up-scaling microfluidic systems for various clinical applications. The advantages and limitations of integrating microfluidic platforms with aptamer development are further addressed, emphasizing areas for future research. We also present a perspective on the future of microfluidic systems and aptamer technologies, highlighting their increasing significance in healthcare and diagnostics.

摘要

适配体是一类合成寡核苷酸,能以高亲和力和特异性与各种靶标结合,这使得它们在诊断、治疗和生物传感领域具有极高价值。微流控平台可以提高适配体筛选的效率和可扩展性,特别是通过指数富集配体系统进化(SELEX)方法的改进。微流控SELEX方法耗时更少、劳动强度更低,包括文库制备、结合、分区和扩增等关键步骤。本综述探讨了微流控技术对基于SELEX的适配体鉴定的贡献,以及用于筛选适配体的条件性SELEX、类SELEX和非SELEX等替代方法,并讨论了过去十年中的关键SELEX步骤。这项工作还研究了用于SELEX的集成微流控系统,重点介绍了条件性SELEX和类SELEX等创新技术。这些进展为使用传统SELEX进行适配体筛选时存在的现有挑战提供了潜在解决方案,尤其是在生物样品方面。还对非SELEX方法的趋势进行了综述和讨论,其中消除了核酸扩增以改进适配体筛选。微流控平台不仅在适配体筛选中展现出多功能性,在各种检测应用中也是如此;它们能够精确控制液体流动,对治疗性适配体的发展至关重要,有助于精确筛选、增强药物递送系统并实现靶向治疗干预。尽管微流控技术的进步有望提升基于适配体的诊断、治疗和生物传感水平,但挑战依然存在,尤其是在扩大微流控系统用于各种临床应用方面。进一步阐述了将微流控平台与适配体开发相结合的优点和局限性,强调了未来研究的方向。我们还对微流控系统和适配体技术的未来发展进行了展望,突出了它们在医疗保健和诊断领域日益重要的地位。

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