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用于检测人雌激素受体α的基于无标记金纳米粒子的比色适体传感器的开发。

Development of a label-free gold nanoparticle-based colorimetric aptasensor for detection of human estrogen receptor alpha.

作者信息

Ahirwar Rajesh, Nahar Pradip

机构信息

Department of Chemical and Systems Biology, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, 110007, India.

CSIR-Institute of Genomics and Integrative Biology, Academy of Scientific and Innovative Research, Delhi, 110007, India.

出版信息

Anal Bioanal Chem. 2016 Jan;408(1):327-32. doi: 10.1007/s00216-015-9090-7. Epub 2015 Oct 17.

Abstract

The increasing demand for easily available and low-cost diagnostics has fuelled the development of aptasensors as platforms for rapid, sensitive, and point-of-care testing of target analytes. Recently, gold nanoparticle (AuNP)-based aptasensors have attracted wide recognition owing to their color transition properties which allow real-time rapid sensing of targets. In this study, we utilized the color transition property of aptamer-functionalized AuNPs to detect and quantify estrogen receptor alpha (ERα), a key biomarker protein in breast cancer. We found that the coating of AuNPs with unmodified ERα-RNA aptamer (GGGGUCAAGGUGACCCC) makes them resistant to salt-induced aggregation. However, addition of ERα to the aptamer-protected AuNPs results in their spontaneous aggregation as evident from a color transition from wine red to deep blue. On the basis of this, we developed an ERα aptasensor, with limits of detection and quantification of 0.64 and 2.16 ng/mL, respectively; the aptasensor can efficiently detect and quantify ERα in a working range of 10 ng/mL-5μg/mL protein. Validation of the aptasensor on cellular extracts of ERα-positive MCF-7 and ERα-deficient MDA-MB-231 breast cancer cells showed a target-selective response in ERα-positive samples but not in cellular extracts of ERα-deficient breast cancer cells. Further, the small size and simple fabrication chemistry of aptamers provide an additional benefit to make the ERα aptasensor a potentially useful and cost-effective tool in point-of-care analyses of ERα.

摘要

对易于获取且低成本诊断方法的需求不断增加,推动了适体传感器的发展,使其成为用于快速、灵敏和即时检测目标分析物的平台。最近,基于金纳米颗粒(AuNP)的适体传感器因其颜色转变特性而受到广泛认可,该特性可实现对目标的实时快速传感。在本研究中,我们利用适体功能化金纳米颗粒的颜色转变特性来检测和定量雌激素受体α(ERα),这是乳腺癌中的一种关键生物标志物蛋白。我们发现,用未修饰的ERα - RNA适体(GGGGUCAAGGUGACCCC)包覆金纳米颗粒可使其抵抗盐诱导的聚集。然而,向适体保护的金纳米颗粒中添加ERα会导致它们自发聚集,从酒红色变为深蓝色的颜色转变就很明显。基于此,我们开发了一种ERα适体传感器,其检测限和定量限分别为0.64和2.16 ng/mL;该适体传感器能够在10 ng/mL - 5μg/mL蛋白质的工作范围内有效检测和定量ERα。在ERα阳性的MCF - 7和ERα缺陷的MDA - MB - 231乳腺癌细胞的细胞提取物上对该适体传感器进行验证,结果显示在ERα阳性样品中呈现目标选择性响应,而在ERα缺陷的乳腺癌细胞的细胞提取物中则没有。此外,适体的小尺寸和简单的制备化学为使ERα适体传感器成为ERα即时分析中潜在有用且具有成本效益的工具提供了额外优势。

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