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AS1411 对铜具有高亲和力;其在用于铜检测的灵敏适体传感器中的应用。

High affinity of AS1411 toward copper; its application in a sensitive aptasensor for copper detection.

机构信息

Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

School of Medicine, St. George's University, Grenada, USA.

出版信息

Anal Biochem. 2019 Jun 15;575:1-9. doi: 10.1016/j.ab.2019.03.016. Epub 2019 Mar 26.

Abstract

AS1411 is a 26-mer G-rich DNA aptamer which has been broadly employed in the field of targeted drug delivery, due to its capability to bind to nucleolin protein on the surface of cancer cells. In this work, it has been shown for the first time that in addition to nucleolin, AS1411 aptamer could bind to copper ions (Cu) with high affinity and selectivity, affecting AS1411 usage in drug delivery systems as a targeting agent. In this study, besides the evaluations of the affinity of AS1411 to different ions and the impact of Cu on targeted drug delivery employed AS1411 aptamer as a targeting agent, a simple and ultra-sensitive fluorescent aptasensor was fabricated for Cu detection through applying AS1411, its complementary strand and magnetic beads coated with streptavidin. Gel Red (GR) was also used as a fluorescent dye. The fabricated aptasensor offered the possibility of condensing samples with different volumes. The detection limit of the sensor was 0.01 μM towards Cu in serum samples. The efficacy of this sensor was further confirmed by comparing Cu levels in serums of healthy people with patients suffering from Wilson's disease, Alzheimer's disease and Diabetes Mellitus using the proposed sensing platform.

摘要

AS1411 是一种 26 个碱基组成的富含 G 的 DNA 适体,由于其能够与癌细胞表面的核仁蛋白结合,因此被广泛应用于靶向药物传递领域。在这项工作中,首次表明除了核仁蛋白外,AS1411 适体还可以与铜离子(Cu)高亲和力和选择性结合,从而影响 AS1411 在作为靶向剂的药物传递系统中的应用。在这项研究中,除了评估 AS1411 与不同离子的亲和力以及 Cu 对靶向药物传递的影响,还将 AS1411 适体用作靶向剂,构建了一种简单且超灵敏的荧光适体传感器,用于通过应用 AS1411、其互补链和链霉亲和素包被的磁性珠来检测 Cu。凝胶红(GR)也被用作荧光染料。所构建的适体传感器提供了对不同体积的样品进行浓缩的可能性。该传感器对血清样品中 Cu 的检测限为 0.01 μM。通过使用该传感平台比较健康人和患有威尔逊病、阿尔茨海默病和糖尿病患者的血清中的 Cu 水平,进一步证实了该传感器的功效。

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