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基于指数富集的配体系统进化(SELEX)技术的快速筛选 CRP 适体的集成微流控系统。

Integrated microfluidic system for rapid screening of CRP aptamers utilizing systematic evolution of ligands by exponential enrichment (SELEX).

机构信息

Department of Engineering Science, National Cheng Kung University, 1, University Road, Tainan, Taiwan.

出版信息

Biosens Bioelectron. 2010 Mar 15;25(7):1761-6. doi: 10.1016/j.bios.2009.12.029. Epub 2009 Dec 28.

Abstract

The systematic evolution of ligands by exponential enrichment (SELEX) is an experimental procedure that allows screening of given molecular targets by desired binding affinities from an initial random pool of oligonucleotides and oligomers. The final products of SELEX are usually referred as aptamers, which are recognized as promising molecules for a variety of biomedical applications. However, SELEX is an iterative process requiring multiple rounds of extraction and amplification that demands significant time and labor. Therefore, this study presents a novel, automatic, miniature SELEX platform. As a demonstration, the rapid screening of C-reactive protein (CRP) aptamers was performed. By utilizing microfluidic technologies and magnetic beads conjugated with CRP, aptamers with a high affinity to CRP were extracted from a random single-strand deoxyribonucleic acid (ssDNA) pool. These aptamers were further amplified by an on-chip polymerase chain reaction (PCR) process. After five consecutive extraction and amplification cycles, a specific aptamer with the highest affinity was screened automatically. The screened aptamers were used as a recognition molecule for the detection of CRP. The developed microsystem demonstrated fast screening of CRP aptamers and can be used as a powerful tool to select analyte-specific aptamers for biomedical applications.

摘要

指数富集配体系统进化(SELEX)是一种实验程序,可通过初始随机寡核苷酸和低聚物库,按照所需的结合亲和力筛选特定的分子靶标。SELEX 的最终产物通常被称为适体,它们被认为是各种生物医学应用的有前途的分子。然而,SELEX 是一个需要多次提取和扩增的迭代过程,需要大量的时间和劳动力。因此,本研究提出了一种新颖的、自动的、微型的 SELEX 平台。作为一个演示,快速筛选 C 反应蛋白(CRP)适体。利用微流控技术和与 CRP 结合的磁珠,从随机单链脱氧核糖核酸(ssDNA)库中提取出与 CRP 具有高亲和力的适体。这些适体通过芯片上的聚合酶链反应(PCR)过程进一步扩增。经过连续五次提取和扩增循环,自动筛选出具有最高亲和力的特定适体。筛选出的适体被用作 CRP 检测的识别分子。所开发的微系统能够快速筛选 CRP 适体,可作为一种强大的工具,用于选择生物医学应用中分析物特异性适体。

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