A Peptide-Functionalized Quantum Dots/MOF Nanosheets Fluorescence Biosensor for Glutathione Sensing and Cellular Imaging

Background Glutathione (GSH), the most abundant cellular non-protein thiol and a central antioxidant, plays essential roles in regulating redox homeostasis, signal transduction, and detoxification processes. Altered GSH levels are closely associated with various pathological conditions, including neurodegenerative disorders, and liver injury . Notably, tumor tissues exhibit markedly elevated GSH concentrations (up to 4-10 times higher than in normal cells), where it promotes tumor proliferation, invasion, and chemoresistance by sustaining a reduced microenvironment, these characteristics make GSH an important biomarker and therapeutic target in oncology. Results We developed a novel "switch-on" near-infrared (NIR) fluorescence nanoprobe, Ag 2 S-CH@Cu-TCPP, for the highly sensitive and selective detection and imaging of GSH. The nanoprobe was constructed by conjugating histidine-rich CH peptides to carboxyl-capped Ag 2 S quantum dots (Ag 2 S QDs), which were then self-assembled onto two-dimensional Cu-TCPP metal-organic framework (MOF) nanosheets via coordination between Cu 2+ and imidazole groups. In this design, Cu-TCPP acts as an efficient quencher for the Ag 2 S-CH fluorescence via a fluorescence resonance energy transfer (FRET) mechanism. Upon encountering GSH, the reduction of Cu 2+ to Cu + disrupts the FRET process, leading to the recovery of the NIR fluorescence signal. This nanoprobe demonstrated a low detection limit of 5.33 μM and a wide linear range from 0.625 to 10 mM for GSH quantification, with excellent selectivity, high reproducibility, and favorable biocompatibility. We successfully applied the platform for monitoring intracellular GSH levels and distinguishing between cancer cells and normal cells based on their differential GSH expression. Significance and Novelty The "switch-on" probe demonstrates excellent biocompatibility and is applicable for cellular imaging. It enables discrimination between normal and cancer cells while supporting semi-quantitative analysis of both endogenous and exogenous GSH. This work provides a novel and reliable strategy for tumor diagnosis and advanced investigations into GSH-related biological processes.