Chimeric Aptamers-Based and MoS2 Nanosheet-Enhanced Label-Free Fluorescence Polarization Strategy for Adenosine Triphosphate Detection
Chimeric Aptamers-Based and MoS2 Nanosheet-Enhanced Label-Free Fluorescence Polarization Strategy for Adenosine Triphosphate Detection
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DOI：
10.1021/acs.analchem.8b04107
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发表时间：
2018-11-20
影响因子：
7.4
通讯作者：
Zhang, Zhi-Qi
中科院分区：
文献类型：
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作者：
Fan, Yao-Yao;Mou, Zhao-Li;Zhang, Zhi-Qi
三磷酸腺苷（ATP）作为一种主要的能量来源，在所有细胞事件的调节中起着独特的作用。检测ATP的必要性需要灵敏和准确的定量分析策略。在这里，我们提出了我们的研究开发一个MoS2纳米片增强的适体传感器，用于荧光偏振检测ATP。设计了一种双功能DNA链，由嵌合适体组成，可以识别和捕获ATP和黄连素（一种荧光增强剂）。在没有ATP的情况下，当引入外切酶I （Exo I）时，与小檗碱结合的DNA链将被水解，从而释放出小檗碱。相反，当ATP存在时，ATP适体折叠成g -四重体结构；因此，该复合物可以抵抗Exo I的降解，以维持小檗碱的荧光检测目的。此外，为了放大荧光偏振（FP）信号，系统中还采用了MoS2纳米片。这种纳米片增强的FP策略简单易行，不需要传统的染料标记DNA链和复杂的操作步骤。所研制的荧光偏振适体传感器对ATP的定量具有较高的灵敏度，检测限为34.4 nM，在缓冲溶液和生物样品中均表现良好。
Adenosine triphosphate (ATP) as a primary energy source plays a unique role in the regulation of all cellular events. The necessity to detect ATP requires sensitive and accurate quantitative analytical strategies. Herein, we present our study of developing a MoS2 nanosheet-enhanced aptasensor for fluorescence polarization-based ATP detection. A bifunctional DNA strand was designed to consist of chimeric aptamers that recognize and capture ATP and berberine, a fluorescence enhancer. In the absence of ATP, the DNA strand bound to berberine will be hydrolyzed when Exonuclease I (Exo I) is introduced, releasing berberine as a result. In contrast, when ATP is present, ATP aptamer folds into a G-quadruplex structure; thus, the complex can resist degradation by Exo I to maintain berberine for fluorescent detection purpose. In addition, to magnify the fluorescence polarization (FP) signal, MoS2 nanosheets were also adopted in the system. This nanosheets-enhanced FP strategy is simple and facile which does not require traditional dye-labeled DNA strands and complex operation steps. The developed fluorescence polarization aptasensor showed high sensitivity for the quantification of ATP with a detection limit of 34.4 nM, performing well both in buffer solution and in biological samples.