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基于适配体的皮摩尔级血小板衍生生长因子在血清中的直接电化学检测。

Aptamer-based electrochemical detection of picomolar platelet-derived growth factor directly in blood serum.

作者信息

Lai Rebecca Y, Plaxco Kevin W, Heeger Alan J

机构信息

Center for Polymers and Organic Solids, Department of Chemistry and Biochemistry, University of California, Santa Barbara, California 93106, USA.

出版信息

Anal Chem. 2007 Jan 1;79(1):229-33. doi: 10.1021/ac061592s.

Abstract

We report an electrochemical, aptamer-based (E-AB) sensor for the detection of platelet-derived growth factor (PDGF) directly in blood serum. The E-AB approach employs alternating current voltammetry to monitor target-induced folding in a methylene blue-modified, PDGF-binding aptamer. The sensor is sensitive, highly selective, and essentially reagentless: we readily detect the BB variant of PDGF at 1 nM directly in undiluted, unmodified blood serum and at 50 pM (1.25 ng/mL) in serum-diluted 2-fold with aqueous buffer. The sensitivity and selectivity achieved by this sensor match or significantly exceed those of the best analogous optical approaches. For example, the detection limit attained in 50% serum is achieved against a >25 million-fold excess of contaminating blood proteins and represents a 4 order of magnitude improvement over the most sensitive optical PDGF aptasensor reported to date. Moreover, the E-AB sensor combines these promising attributes in a platform that is reusable, label-free, and electronic. Given these advantages, E-AB sensors appear well suited for implementation in portable microdevices directed at the direct detection of proteins and small molecules in complex, largely unprocessed clinical samples.

摘要

我们报道了一种基于适体的电化学(E-AB)传感器,用于直接检测血清中的血小板衍生生长因子(PDGF)。E-AB方法采用交流伏安法来监测在亚甲蓝修饰的、与PDGF结合的适体中目标诱导的折叠。该传感器灵敏、高度选择性且基本无需试剂:我们能够直接在未稀释、未处理的血清中轻松检测到浓度为1 nM的PDGF BB变体,在用水性缓冲液2倍稀释的血清中能检测到浓度为50 pM(1.25 ng/mL)的该变体。该传感器实现的灵敏度和选择性与最佳的类似光学方法相当或显著超过它们。例如,在50%血清中达到的检测限是在超过2500万倍过量的污染血液蛋白存在下实现的,相较于迄今报道的最灵敏的光学PDGF适体传感器,提高了4个数量级。此外,E-AB传感器在一个可重复使用、无标记且电子化的平台中结合了这些有前景的特性。鉴于这些优势,E-AB传感器似乎非常适合应用于旨在直接检测复杂的、基本未处理的临床样本中的蛋白质和小分子的便携式微型设备。

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