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基于新筛选的适体的用于高度特异性检测 ATP 的荧光适体传感器。

Fluorescent Aptasensor for Highly Specific Detection of ATP Using a Newly Screened Aptamer.

机构信息

The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.

Medical College, Nantong University, Nantong 226007, China.

出版信息

Sensors (Basel). 2022 Mar 22;22(7):2425. doi: 10.3390/s22072425.

Abstract

Owing to the significant roles of adenosine triphosphate (ATP) in diverse biological processes, ATP level is used to research and evaluate the physiological processes of organisms. Aptamer-based biosensors have been widely reported to achieve this purpose, which are superior in their flexible biosensing mechanism, with a high sensitivity and good biocompatibility; however, the aptamers currently used for ATP detection have a poor ability to discriminate ATP from adenosine diphosphate (ADP) and adenosine monophosphate (AMP). Herein, an ATP-specific aptamer was screened and applied to construct a fluorescent aptasensor for ATP by using graphene oxide (GO) and strand displacement amplification (SDA). The fluorescence intensity of the sensor is linearly related to the concentration of ATP within 0.1 μM to 25 μM under optimal experimental conditions, and the detection limit is 33.85 nM. The biosensor exhibits a satisfactory specificity for ATP. Moreover, the experimental results indicate that the biosensor can be applied to determine the ATP in human serum. In conclusion, the screened aptamer and the biosensor have promising applications in the determination of the real energy charge level and ATP content in a complex biological system.

摘要

由于三磷酸腺苷(ATP)在多种生物过程中具有重要作用,因此 ATP 水平被用于研究和评估生物体的生理过程。基于适配体的生物传感器已被广泛报道可实现这一目的,其具有灵活的生物传感机制,灵敏度高,生物相容性好;然而,目前用于检测 ATP 的适配体对 ATP 与二磷酸腺苷(ADP)和一磷酸腺苷(AMP)的区分能力较差。在此,筛选出一种对 ATP 具有特异性的适配体,并通过氧化石墨烯(GO)和链置换扩增(SDA)将其应用于构建用于检测 ATP 的荧光适配体传感器。在最佳实验条件下,传感器的荧光强度与 0.1 μM 至 25 μM 范围内 ATP 浓度呈线性关系,检测限为 33.85 nM。该生物传感器对 ATP 具有令人满意的特异性。此外,实验结果表明,该生物传感器可用于测定人血清中的 ATP。总之,筛选出的适配体和生物传感器有望应用于复杂生物体系中真实能量电荷水平和 ATP 含量的测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0921/9003339/a62d56fee564/sensors-22-02425-g001.jpg

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