Bai Cheng, Meng Xin, Wen Kechun, Citartan Marimuthu, Wang Chaohui, Yu Shifeng, Lin Qiao
Department of Mechanical Engineering, Columbia University, New York, NY 10027, USA.
State Key Laboratory for Manufacturing Systems Engineering, Xi'an Jiaotong University, Xi'an 710049, Shaanxi, People's Republic of China.
Microfluid Nanofluidics. 2022 Jun;26(6). doi: 10.1007/s10404-022-02548-w. Epub 2022 May 17.
Aptamers are synthetic single-stranded nucleic acid molecules that bind to biochemical targets with high affinity and specificity. The method of systematic evolution of ligands by exponential enrichment (SELEX) is widely used to isolate aptamers from randomized oligonucleotides. Recently, microfluidic technology has been applied to improve the efficiency and reduce the cost in SELEX processes. In this work, we present an approach that exploits surface acoustic waves to improve the affinity selection process in microfluidic SELEX. Acoustic streaming is used to enhance the interactions of the solution-based oligonucleotide molecules with microbead-immobilized target molecules, allowing the identification of high-affinity aptamer candidates in a more efficient manner. For demonstration, a DNA aptamer is isolated within three rounds of selection in 5 h to specifically bind to immunoglobulin E, a representative target protein, with an equilibrium dissociation constant of approximately 22.6 nM.
适配体是一种合成的单链核酸分子,能以高亲和力和特异性结合生化靶标。指数富集配体系统进化技术(SELEX)被广泛用于从随机寡核苷酸中分离适配体。最近,微流控技术已被应用于提高SELEX过程的效率并降低成本。在这项工作中,我们提出了一种利用表面声波来改进微流控SELEX中亲和力选择过程的方法。声流被用于增强基于溶液的寡核苷酸分子与固定在微珠上的靶标分子之间的相互作用,从而能够更高效地鉴定高亲和力适配体候选物。为进行演示,在5小时内经过三轮筛选分离出一种DNA适配体,它能特异性结合免疫球蛋白E(一种代表性靶标蛋白),平衡解离常数约为22.6 nM。