Fan Yao-Yao, Deng Xu, Wang Man, Li Jun, Zhang Zhi-Qi
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, China.
Talanta. 2020 Nov 1;219:121349. doi: 10.1016/j.talanta.2020.121349. Epub 2020 Jul 7.
Adenosine triphosphate (ATP) is the main energy currency of life that plays a vital role in supporting physiological activities in living organisms, including humans. Therefore, accurate and sensitive detection of ATP concentration is necessary in biochemical research and clinical diagnosis. Herein, a ratiometric fluorescence aptasensor was developed for ATP detection. A dual-function DNA strand comprising an ATP-binding aptamer (ABA) and berberine-binding aptamer (BBA) was designed and optimized, in which ABA can capture ATP and thioflavin T (ThT), whereas BBA can capture berberine. Interestingly, the fluorescence intensity of both berberine and ThT were enhanced as they were captured by this dual-function DNA strand. In the presence of ATP, the ABA on the 3'-end of the DNA bound specifically to its target, causing ThT release and a significant drop in ThT fluorescence. However, ATP had no significant effect on the interaction between berberine and DNA, remaining the enhanced fluorescence intensity of berberine stable. Based on this interesting phenomenon, a ratiometric fluorescence sensor was constructed that used the enhanced fluorescence intensity of berberine as reference to measure the fluorescence intensity of ThT for ATP detection. This ratiometric fluorescence strategy had excellent selectivity and high sensitivity towards ATP with a detection limit (3σ) as low as 24.8 nM. The feasibility of application of this method in biological samples was evaluated in human serum and urine samples, where it exhibited a good detection performance.
三磷酸腺苷（ATP）是生命的主要能量货币，在支持包括人类在内的生物体内的生理活动中起着至关重要的作用。因此，在生化研究和临床诊断中，准确、灵敏地检测ATP浓度是必要的。在此，开发了一种用于ATP检测的比率荧光适配体传感器。设计并优化了一种包含ATP结合适配体（ABA）和黄连素结合适配体（BBA）的双功能DNA链，其中ABA可以捕获ATP和硫黄素T（ThT），而BBA可以捕获黄连素。有趣的是，黄连素和ThT被这种双功能DNA链捕获时，它们的荧光强度都会增强。在存在ATP的情况下，DNA 3'端的ABA特异性结合其靶标，导致ThT释放，ThT荧光显著下降。然而，ATP对黄连素与DNA之间的相互作用没有显著影响，黄连素的荧光强度仍保持增强且稳定。基于这一有趣现象，构建了一种比率荧光传感器，该传感器以黄连素增强的荧光强度为参考，测量ThT的荧光强度以检测ATP。这种比率荧光策略对ATP具有出色的选择性和高灵敏度，检测限（3σ）低至24.8 nM。在人血清和尿液样本中评估了该方法在生物样品中应用的可行性，其表现出良好的检测性能。
