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A sensitive and rapid detection of glutathione based on a fluorescence-enhanced "turn-on" strategy
Qianru Ma1, Minning Wang1, Huahuan Cai1
1Department of Chemistry, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian 361005, P. R. China. stacyliu@xmu.edu.cn and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian 361005, P. R. China and The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen, Fujian 361005, P. R. China.
Abstract:
Glutathione (GSH) plays important roles in the human body including protecting cells from oxidative damages and maintaining cellular redox homeostasis. Thus, developing a fast and sensitive method for detecting GSH levels in living bodies is of great importance. Many methods have been developed and used for GSH detection, such as high-performance liquid chromatography, capillary electrophoresis, and fluorescence resonance energy-based methods. However, these methods often lack sensitivity as well as efficiency. Herein, a rapid and sensitive method for glutathione detection was developed based on a fluorescence-enhanced "turn-on" strategy. In this study, a unique and versatile bifunctional linker 3-[(2-aminoethyl) dithio]propionic acid (AEDP)-modified gold nanoparticle (Au@PLL-AEDP-FITC) probe was designed for the simple, highly sensitive intracellular GSH detection, combined with the FRET technique. In the presence of GSH, the disulfide bonds of AEDP on Au@PLL-AEDP-FITC were broken through competition with GSH, and FITC was separated from gold nanoparticles, making the fluorescence signal switch to the "turn on" state. A change in the fluorescence signal intensity has a great linear positive correlation with GSH concentration, in the linear range from 10 nM to 180 nM (R2 = 0.9948), and the limit of detection (LOD) of 3.07 nM, which was lower than other reported optical nanosensor-based methods. Au@PLL-AEDP-FITC also has great selectivity for GSH, making it promising for application in complex biological systems. The Au@PLL-AEDP-FITC probe was also successfully applied in intracellular GSH imaging in HeLa cells with confocal microscopy. In short, the Au@PLL-AEDP-FITC probe-based fluorescence-enhanced "turn-on" strategy is a sensitive, fast, and effective method for GSH detection as compared with other methods. It can be applied in complex biological systems such as cell systems, with promising biological-medical applications in the future.
