MOF-Wrapped Cu Nanocluster-Based Fluorescent Sensor Array for Phosphates High-Throughput Analysis and Their Enzymatic Hydrolysis Monitoring

Phosphate plays a crucial role in various physiological and pathological processes; however, effective strategies for analyzing diverse phosphates and monitoring phosphate hydrolysis remain limited. Herein, we develop a metal–organic framework wrapped Cu nanoclusters-based (CuNCs@MOF) fluorescent sensor array for high-throughput discrimination of phosphates and real-time monitoring of their enzymatic hydrolysis. Leveraging the confinement effect of zinc-based MOFs, the encapsulated CuNCs exhibited reduced intramolecular vibration and rotation, resulting in enhanced luminescence performance. Due to the distinct affinities of various phosphates toward zinc ions, the sensor array generates differentiated fluorescence responses for seven phosphates, including adenosine triphosphate (ATP), cytidine triphosphate, uridine triphosphate, adenosine diphosphate, adenosine monophosphate, pyrophosphate (PPi), and inorganic phosphate. Subsequently, the fluorescence response signals are processed using the pattern recognition algorithm to generate the unique fingerprint spectra of each phosphate. We demonstrate that the sensor array can not only identify seven distinct phosphates at concentrations as low as 5 μM and effectively distinguish between their mixtures but also exhibit excellent anti-interference capability and enable dynamic monitoring of ATP and PPi hydrolysis. Notably, the sensor array achieved outstanding performance in complex real-world samples and cellular-level analyses, demonstrating 100% accuracy in blind sample identification. Overall, this work develops a strategy for high-throughput discrimination of phosphates and real-time monitoring of their enzymatic hydrolysis.