Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China; South China Academy of Advanced Optoelectronics, South China Normal University, Guangzhou, 510006, PR China.
Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China.
Anal Chim Acta. 2022 Oct 16;1230:340414. doi: 10.1016/j.aca.2022.340414. Epub 2022 Sep 19.
Assays utilizing in situ fluorogenic reactions provide a simple and convenient alternative approach for the detection of biological molecules and activities. In this work, a novel ratiometric fluorescent probe based on in situ fluorogenic reaction is explored and developed for alkaline phosphatase (ALP) activity sensing. An intriguing fluorogenic reaction between 2,3-diaminonaphthalene (2,3-DAN) and ascorbic acid (AA) in alkaline aqueous solutions could generate the fluorescent quinoxaline derivative. The resultant quinoxaline emits intense yellow fluorescence, differing from the blue fluorescence of 2,3-DAN. Thus, a ratiometric fluorescent probe based on this fluorogenic reaction is constructed for ALP activity sensing, combining with ALP-triggered hydrolysis of AA2P into AA. Meanwhile, the addition of copper(II) acetate into the reaction system largely improves reaction rate and efficiency. This sensing strategy shows high sensitivity for ALP activity with detection limit of 0.08 U/L, and excellent selectivity towards ALP out of various interferences. This method is extended to human salivary ALP detection. The present method provides a simple and reliable alternative for the detection of ALP activity and has the potential for clinical applications. It also shows a feasible way to construct ratiometric fluorescent methods.
利用原位荧光反应的测定方法为生物分子和活性的检测提供了一种简单便捷的替代方法。在这项工作中，探索并开发了一种基于原位荧光反应的新型比率荧光探针，用于碱性磷酸酶（ALP）活性传感。在碱性水溶液中，2,3-二氨基萘（2,3-DAN）和抗坏血酸（AA）之间的有趣荧光反应可以生成荧光喹喔啉衍生物。所得喹喔啉发出强烈的黄色荧光，与 2,3-DAN 的蓝色荧光不同。因此，构建了基于该荧光反应的比率荧光探针，用于 ALP 活性传感，结合 ALP 触发的 AA2P 水解为 AA。同时，向反应体系中加入醋酸铜（II）可大大提高反应速率和效率。该传感策略对 ALP 活性具有高灵敏度，检测限为 0.08 U/L，对各种干扰物具有优异的选择性。该方法被扩展到人唾液 ALP 的检测。本方法为 ALP 活性的检测提供了一种简单可靠的替代方法，具有临床应用的潜力。它还为构建比率荧光方法提供了一种可行的途径。
