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使用氧化石墨烯包覆的磁性纳米颗粒在芯片上筛选腺苷适配体。

On-chip selection of adenosine aptamer using graphene oxide-coated magnetic nanoparticles.

作者信息

Lim Hosub, Chang Junhyuck, Kim Kyung-Il, Moon Youngkwang, Lee Saebom, Lee Byoungsang, Lee Jung Heon, Lee Jinkee

机构信息

School of Advanced Materials Science and Engineering, Sungkyunkwan University, Suwon, Republic of Korea.

School of Mechanical Engineering, Sungkyunkwan University, Suwon, Republic of Korea.

出版信息

Biomicrofluidics. 2022 Jul 28;16(4):044102. doi: 10.1063/5.0095419. eCollection 2022 Jul.

Abstract

Systematic evolution of ligands by exponential enrichment (SELEX) is a method that is generally used for developing aptamers, which have arisen the promising alternatives for antibodies. However, conventional SELEX methods have limitations, such as a limited selection of target molecules, time-consuming and complex fabrication processes, and labor-intensive processes, which result in low selection yields. Here, we used (i) graphene oxide (GO)-coated magnetic nanoparticles in the selection process for separation and label-free detection and (ii) a multilayered microfluidic device manufactured using a three-dimensionally printed mold that is equipped with automated control valves to achieve precise fluid flows. The developed on-chip aptamer selection device and GO-coated magnetic nanoparticles were used to screen aptamer candidates for adenosine in eight cycles of the selection process within approximately 2 h for each cycle. Based on results from isothermal titration calorimetry, an aptamer with a dissociation constant of 18.6 ± 1.5 M was selected. Therefore, the on-chip platform based on GO-coated magnetic nanoparticles provides a novel label-free screening technology for biosensors and micro/nanobiotechnology for achieving high-quality aptamers.

摘要

指数富集的配体系统进化技术(SELEX)是一种通常用于开发适体的方法,适体已成为抗体的有前景的替代物。然而,传统的SELEX方法存在局限性,例如目标分子的选择有限、耗时且复杂的制备过程以及劳动强度大的过程,这些导致选择产率较低。在此,我们在选择过程中使用了(i)氧化石墨烯(GO)包覆的磁性纳米颗粒进行分离和无标记检测,以及(ii)使用三维打印模具制造的多层微流控装置,该装置配备自动控制阀以实现精确的流体流动。所开发的芯片上适体选择装置和GO包覆的磁性纳米颗粒用于在大约2小时的每个选择过程周期内进行八个周期的腺苷适体候选物筛选。基于等温滴定量热法的结果,选择了一种解离常数为18.6±1.5 M的适体。因此,基于GO包覆磁性纳米颗粒的芯片上平台为生物传感器和微/纳米生物技术提供了一种新颖的无标记筛选技术,以获得高质量的适体。

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