期刊
BIOSENSORS & BIOELECTRONICS
卷 63, 期 -, 页码 311-316出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2014.07.064
关键词
Aptasensor; ATP; Fluorescence; Label-free; Enzyme catalysis
类别
资金
- National Natural Science Foundation of China [21347008, 21175086]
- Natural Science Foundation of Shanxi Province [2010021010-2]
- China Postdoctoral Science Foundation [201104651, 20100471581]
- The Faculty Research and Professional Development (FRPD) Award
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.
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