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基于纳流控电化学平台结合 G-四链体适体对钾离子和三磷酸腺苷的定量检测。

Quantitative detection of potassium ions and adenosine triphosphate via a nanochannel-based electrochemical platform coupled with G-quadruplex aptamers.

机构信息

Jiangsu Province Hi-Tech Key Laboratory for Bio-medical Research, Suzhou Research Institute of Southeast University, School of Chemistry and Chemical Engineering, Southeast University , Nanjing 210096, P. R. China.

出版信息

Anal Chem. 2014 Nov 4;86(21):10741-8. doi: 10.1021/ac502752g. Epub 2014 Oct 21.

Abstract

The development of synthetic nanopores and nanochannels that mimick ion channels in living organisms for biosensing applications has been, and still remains, a great challenge. Although the biological applications of nanopores and nanochannels have achieved considerable development as a result of nanotechnology advancements, there are few reports of a facile way to realize those applications. Herein, a nanochannel-based electrochemical platform was developed for the quantitative detection of biorelated small molecules such as potassium ions (K(+)) and adenosine triphosphate (ATP) in a facile way. For this purpose, K(+) or ATP G-quadruplex aptamers were covalently assembled onto the inner wall of porous anodic alumina (PAA) nanochannels through a Schiff reaction between -CHO groups in the aptamer and amino groups on the inner wall of the PAA nanochannels under mild reaction conditions. Conformational switching of the aptamers confined in the nanochannels occurs in the presence of the target molecules, resulting in increased steric hindrance in the nanochannels. Changes in steric hindrance in the nanochannels were monitored by the anodic current of indicator molecules transported through the nanochannels. As a result, quantitative detection of K(+) and ATP was realized with a concentration ranging from 0.005 to 1.0 mM for K(+) and 0.05 to 10.0 mM for ATP. The proposed platform displayed significant selectivity, good reproducibility, and universality. Moreover, this platform showed its potential for use in the detection of other aptamer-based analytes, which could promote its development for use in biological detection and clinical diagnosis.

摘要

用于生物传感应用的模拟生物体内离子通道的合成纳米孔和纳米通道的发展一直是一个巨大的挑战。尽管纳米技术的进步使得纳米孔和纳米通道的生物应用取得了相当大的发展,但实现这些应用的简便方法却很少有报道。在此,我们开发了一种基于纳米通道的电化学平台,用于简便地定量检测生物相关小分子,如钾离子 (K(+)) 和三磷酸腺苷 (ATP)。为此,通过 K(+) 或 ATP G-四链体适体与 PAA 纳米通道内壁上的氨基之间的席夫反应,将 -CHO 基团共价组装到多孔阳极氧化铝 (PAA) 纳米通道的内壁上。在温和的反应条件下,在存在靶分子的情况下,适体在纳米通道中发生构象转换,导致纳米通道中的空间位阻增加。纳米通道中空间位阻的变化通过通过纳米通道传输的指示剂分子的阳极电流进行监测。结果,实现了 K(+) 和 ATP 的定量检测,K(+) 的浓度范围为 0.005 至 1.0 mM,ATP 的浓度范围为 0.05 至 10.0 mM。所提出的平台表现出显著的选择性、良好的重现性和通用性。此外,该平台在检测其他基于适体的分析物方面显示出了潜力,这将促进其在生物检测和临床诊断中的应用。

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