Integrated microfluidic system for rapid screening of CRP aptamers utilizing systematic evolution of ligands by exponential enrichment (SELEX)
Integrated microfluidic system for rapid screening of CRP aptamers utilizing systematic evolution of ligands by exponential enrichment (SELEX)
复制标题
DOI：
10.1016/j.bios.2009.12.029
复制
发表时间：
2010-03-15
影响因子：
12.6
通讯作者：
Lee, Gwo-Bin
中科院分区：
文献类型：
--
作者：
Huang, Chao-June;Lin, Hsin-I;Lee, Gwo-Bin
通过指数富集的配体系统进化 (SELEX) 是一种实验程序，允许通过所需的结合亲和力从寡核苷酸和寡聚物的初始随机池中筛选给定的分子靶标。 SELEX 的最终产品通常称为适体，被认为是多种生物医学应用的有前途的分子。然而，SELEX 是一个迭代过程，需要多轮提取和放大，需要大量时间和劳动力。因此，本研究提出了一种新颖的、自动化的、微型的SELEX平台。作为示范，我们对 C 反应蛋白 (CRP) 适体进行了快速筛选。通过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。这些适体通过片上聚合酶链式反应（PCR）过程进一步扩增。经过五个连续的提取和扩增循环后，自动筛选出具有最高亲和力的特定适配体。筛选出的适配体作为识别分子用于CRP的检测。开发的微系统证明了 CRP 适体的快速筛选，并且可以用作选择用于生物医学应用的分析物特异性适体的强大工具。 (C) 2009 Elsevier B.V. 保留所有权利。
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. (C) 2009 Elsevier B.V. All rights reserved.