In this experimental research, a label-free sensing strategy is developed and employed to detect adenosine triphosphate with utilization of aptamers, including exonuclease I and SYBR Gold. The conformation of aptamers bonding to the specific target molecule (ATP) is transformed into an antiparallel G-quadruplex structure from a random coil. Afterwards, considering the unfolded aptamers are the preferred substrates for exonuclease I, the addition of exonuclease I is used so as to digest unfolded aptamers in the mixture in a selective manner. In the follow-up study, in order to strengthen the fluorescence intensity, SYBR Gold is applied as a fluorescent probe. The aptasensor presents the features of high selectivity against adenosine triphosphate and the low detecting limit of concentrations (39.2 nM). In order to verify the validation of experimental procedures and the practical application of the aptasensor, the detection of adenosine triphosphate for human serum samples is performed with satisfactory success. The recovery result with the range of 93.8%–108.1% is desirable and suggests that the designed approach is applicable. The outcomes of the cellular adenosine triphosphate assay manifest that the level of adenosine triphosphate concentrations in cell extracts can be monitored without the interference of other substances in the cells. Subject to its advantageous benefits (cost-effective, easiness, rapidity, and extraordinary selectivity), the designed approach has a promising implication for adenosine triphosphate detection in the research domain of bioanalytical science and biology.
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