Integrated Microfluidic Isolation of Aptamers Using Electrophoretic Oligonucleotide Manipulation
Integrated Microfluidic Isolation of Aptamers Using Electrophoretic Oligonucleotide Manipulation
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DOI：
10.1038/srep26139
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发表时间：
2016-05-24
影响因子：
4.6
通讯作者：
Lin, Qiao
中科院分区：
文献类型：
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作者：
Kim, Jinho;Olsen, Timothy R.;Lin, Qiao
我们提出了一种微流控的方法，通过指数富集配体的系统进化（SELEX）集成分离的DNA适体。该方法采用了基于微珠的协议的亲和选择和扩增的目标结合寡核苷酸的过程中，和电泳DNA操作方案的耦合这些过程中，需要发生在不同的缓冲液。这实现了SELEX的完全微流体整合，从而能够在大幅减少的时间内高效分离适体，并最大限度地减少生物材料的消耗。该方法本身还通过允许相对于表面固定的或溶液携带的靶标选择适体来提供广泛的靶标适用性，潜在地允许适体被开发为用于广泛靶标的容易获得的亲和试剂。我们证明了这种方法在两种不同的程序上的实用性，分别用于分离针对表面固定化蛋白（免疫球蛋白E）和溶液相小分子（在葡萄糖存在下的双硼酸）的适体。在两种情况下，在约10小时的总处理时间内，在三轮SELEX中分离适体候选物。
We present a microfluidic approach to integrated isolation of DNA aptamers via systematic evolution of ligands by exponential enrichment (SELEX). The approach employs a microbead-based protocol for the processes of affinity selection and amplification of target-binding oligonucleotides, and an electrophoretic DNA manipulation scheme for the coupling of these processes, which are required to occur in different buffers. This achieves the full microfluidic integration of SELEX, thereby enabling highly efficient isolation of aptamers in drastically reduced times and with minimized consumption of biological material. The approach as such also offers broad target applicability by allowing selection of aptamers with respect to targets that are either surface-immobilized or solution-borne, potentially allowing aptamers to be developed as readily available affinity reagents for a wide range of targets. We demonstrate the utility of this approach on two different procedures, respectively for isolating aptamers against a surface-immobilized protein (immunoglobulin E) and a solution-phase small molecule (bisboronic acid in the presence of glucose). In both cases aptamer candidates were isolated in three rounds of SELEX within a total process time of approximately 10 hours.