INTEGRATED MICROFLUIDIC SELEX USING FREE SOLUTION ELECTROKINETICS.
INTEGRATED MICROFLUIDIC SELEX USING FREE SOLUTION ELECTROKINETICS.
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DOI：
10.1149/2.0191705jes
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发表时间：
2017
影响因子：
3.9
通讯作者：
Lin Q
中科院分区：
文献类型：
--
作者：
Olsen TR;Tapia-Alveal C;Yang KA;Zhang X;Pereira LJ;Farmakidis N;Pei R;Stojanovic MN;Lin Q
通过指数富集的配体系统进化（SELEX）提供了分离称为适体的亲和寡核苷酸的强有力的方法，所述亲和寡核苷酸然后可以用于从药物递送到生物传感的广泛应用中。然而，传统的SELEX方法依赖于劳动密集型和耗时的台式操作。提出了一种简化的微流体方法，其允许通过将基于珠的生化反应与自由溶液电动寡核苷酸转移相结合来整合SELEX的亲和选择和扩增阶段以用于分离靶结合寡核苷酸。自由溶液电动力学允许亲和选择和扩增的偶联用于闭环寡核苷酸富集，而不需要离线过程、流动处理组件或凝胶组件，而基于珠的选择和扩增允许试剂和反应产物的有效操作，从而实现芯片上环闭合和整个SELEX过程的整合。因此，该方法能够使用简单的转移过程进行多轮寡核苷酸富集，同时保持高水平的设备集成，正如在大约4小时的处理时间内分离针对蛋白质靶点（伊加）的适体池所证明的那样，其结合亲和力显着高于起始库。
Systematic evolution of ligands by exponential enrichment (SELEX) offers a powerful method to isolate affinity oligonucleotides known as aptamers, which can then be used in a wide range of applications from drug delivery to biosensing. However, conventional SELEX methods rely on labor intensive and time consuming benchtop operations. A simplified microfluidic approach is presented which allows integration of the affinity selection and amplification stages of SELEX for the isolation of target-binding oligonucleotides by combining bead-based biochemical reactions with free solution electrokinetic oligonucleotide transfer. Free solution electrokinetics allows coupling of affinity selection and amplification for closed loop oligonucleotide enrichment without the need for offline processes, flow handling components or gel components, while bead based selection and amplification allow efficient manipulation of reagents and reaction products thereby realizing on-chip loop closure and integration of the entire SELEX process. Thus the approach is capable of multi-round enrichment of oligonucleotides using simple transfer processes while maintaining a high level of device integration, as demonstrated by the isolation of an aptamer pool against a protein target (IgA) with significantly higher binding affinity than the starting library in approximately 4 hours of processing time.
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影响因子：
4.6
作者：
Birch CM;Hou HW;Han J;Niles JC
通讯作者：
Niles JC
影响因子：
14.9
作者：
Geiger, A;Burgstaller, P;Famulok, M
通讯作者：
Famulok, M
影响因子：
4.6
作者：
Kim, Jinho;Olsen, Timothy R.;Lin, Qiao
通讯作者：
Lin, Qiao
影响因子：
15.9
作者：
McNamara, James O., II;Kolonias, Despina;Gilboa, Eli
通讯作者：
Gilboa, Eli
DOI：
10.1073/pnas.1009331107
发表时间：
2010-08-31
影响因子：
11.1
作者：
Cho, Minseon;Xiao, Yi;Soh, H. Tom
通讯作者：
Soh, H. Tom