An exonuclease I-based label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection with a wide concentration range
An exonuclease I-based label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection with a wide concentration range
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基于核酸外切酶 I 的无标记荧光适体传感器，用于宽浓度范围的三磷酸腺苷 (ATP) 检测
DOI：
10.1016/j.bios.2014.07.064
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发表时间：
2015-01-15
影响因子：
12.6
通讯作者：
Wang, Gufeng
中科院分区：
文献类型：
--
作者：
Wei, Yanli;Chen, Yanxia;Wang, Gufeng
建立了一种新的基于核酸适体的ATP无标记检测方法。该检测采用了一种新的适体/荧光探针系统，该系统在与ATP分子结合时显示出对核酸外切酶I（Exo I）消化的抗性。在不存在ATP的情况下，ATP结合适体（ATP-适体）和DNA结合染料小檗碱之间的复合物在添加核酸外切酶I时被消化，导致小檗碱释放到溶液中，并因此淬灭小檗碱荧光。在ATP存在下，ATP结合适体折叠成G-四链体结构，其对Exo I消化具有抗性。因此，小檗碱在G-四链体结构中被保护，并且观察到高荧光强度。因此，基于荧光信号变化，开发了ATP的无标记荧光测定。考察了ATP结合适体浓度、反应时间、温度和Exo I浓度等因素对ATP分析的影响。在最佳条件下，该传感系统的荧光强度显示出响应ATP在宽范围内高达17.5 mM，检测限为140 nM。(C)2014爱思唯尔有限公司版权所有。
A novel aptamer-based label-free assay for sensitive and selective detection of ATP was developed. This assay employs a new aptamer/fluorescent probe system that shows resistance to exonuclease I (Exo I) digestion upon binding to ATP molecules. In the absence of ATP, the complex between the ATP-binding aptamer (ATP-aptamer) and a DNA binding dye, berberine, is digested upon the addition of exonuclease I, leading to the release of berberine into solution and consequently, quenched berberine fluorescence. In the presence of ATP, the ATP-binding aptamer folds into a G-quadruplex structure that is resistant to Exo I digestion. Accordingly, berberine is protected in the G-quadruplex structure and high fluorescence intensity is observed. As such, based on the fluorescence signal change, a label-free fluorescence assay for ATP was developed. Factors affecting the analysis of ATP including the concentration of ATP-binding aptamer, reaction time, temperature and the concentration of Exo I were comprehensively investigated. Under optimal conditions, the fluorescence intensity of the sensing system displayed a response for ATP in a wide range up to 17.5 mM with a detection limit of 140 nM. (C) 2014 Elsevier B.V. All rights reserved.