Improved Affinity: A Customized Fluorescent Probe for the Rapid Detection of Butyrylcholinesterase

This article presents the distinctive butyrylcholinesterase (BChE) fluorescent probe P5, designed via a targeting-site method. This method was proposed to enhance the affinity of the probe for BChE by targeting the peripheral anionic site (PAS) of BChE. By mimicking the natural substrate butyrylcholine, the structure of the probe was optimized by introducing a positive charge. Fluorescent probe P5, selected from a series of designed fluorescent probes P1–P6, exhibited excellent affinity and specificity towards BChE, enabling rapid detection within 5 min with a low detection limit of 16.7 ng/mL. Furthermore, this probe can distinguish between normal cells and Alzheimer’s disease (AD) model cells, and demonstrated good imaging results in a P12 cell AD model. The results of this study indicate that this novel fluorescent probe could serve as a promising tool for the rapid detection of BChE and accurate AD diagnosis.