A supramolecular gel‐based protocol for the detection of α‑glycosidases for screening potential drugs

α‐Glycosidases are carbohydrate‐digesting enzymes that catalyze the hydrolysis of α‑1,4‑glycopyranoside bonds from oligosaccharides and disaccharides. α‑Glucosidase is an important biomarker for the diagnosis of type‑II diabetes, Azoospermia and Pompe diseases. Additionally, mutations in α‑galactosidase lead to Fabry disease. Inhibitors targeting these enzymes are prescribed as anti‐diabetic medications and as effective chaperones for Fabry disease. Comprehending the function and regulation of α‑glycosidases requires accurate quantification methods. In this work, we highlight the design of a simple luminescent ‘turn‐on’ assay for sensing these two α‐glycosidases in a supramolecular TbCh hydrogel matrix using 1‐α‐glycosides as pro‑sensitizers. Our protocol offers a simpler and cost‐effective method for selectively sensing the detection limit of the subnanomolar range of α‐glycosidases. Importantly, the developed enzyme sensors functioned as a platform for rapid screening of drug molecules based on their inhibition potency. Therefore, the protocol is useful for facilitating the development of therapeutics and diagnostics for this important class of enzymes.