Copper(II) complex as a turn on fluorescent sensing platform for acetylcholinesterase activity with high sensitivity

Acetylcholinesterase (AChE) is an important enzyme associated with many nervous diseases, demonstrating the great need for smarter sensing platform with improved sensitivity, selectivity and simplified operation. A "turn on" fluorometric assay is described herein for AChE activity detection, according to the specific enzyme catalyzed reaction of acetylcholine (ATCh) by AChE, which generates thiocholine (TCh) as the product. The well-designed fluorescent probe HBTP possesses ESIPT (Excited State Intramolecular Proton Transfer) nature, leading to a larger Stokes shift, which could be quenched upon coordination with Cu²⁺. The fluorescence-silent HBTP-Cu²⁺ complex could be broken by TCh generated from reaction of ATCh with AChE, giving rise to HBTP release which originates from competitive coordination of TCh with Cu²⁺. This complex probe HBTP-Cu²⁺ offers a limit detection as low as 0.02 mU mL^-1, which is lower than most reported literatures. Furthermore, both HBTP-Cu²⁺ and HBTP show little toxicity to live cells and is available in visualizing cellular AChE activity.