Dextran-assisted ultrasonic exfoliation of two-dimensional metal-organic frameworks to evaluate acetylcholinesterase activity and inhibitor screening

Acetylcholinesterase (AChE) is regarded as a biomarker of Alzheimer's disease (AD), and its inhibitors show great potential in AD therapy as AChE can increase the neurotoxicity of the amyloid component that induces AD. Because of this, it is crucial and significant to develop a simple and highly sensitive strategy to monitor AChE levels and screen highly efficient AChE inhibitors. Herein, we synthesize an ultrathin two-dimensional (2D) metal-organic framework (MOF) based on copper-catecholate (Cu-CAT) via dextran assisted ultrasound exfoliation, followed by construction of a sensitive sensor for the monitoring AChE and screening of its inhibitors. By adding AChE, the acetylthiocholine (ATCh) substrate is hydrolyzed to be thiocholine (TCh), which decreases the peroxidase-like activity of Cu-CAT nanosheets (Cu-CAT NSs), impairing the signal reaction of 3,3',5,5'-tetramethylbenzidine (TMB) to oxidized-TMB (ox-TMB). In the presence of an AChE inhibitor, the signal can be gradually restored. The newly developed sensor shows high sensitivity and selectivity for AChE and huperzine A (HA, an effective drug for AD, an acetylcholine receptor antagonist), as well as for AD drug discovery from traditional Chinese herbs. The limit of detection of the sensor for AChE is 0.01 mU mL-1 and the average IC50 value of HA is 30.81 nM under the optimal of catalysis conditions. Compared with the 3D bulk Cu-CAT, the current 2D Cu-CAT NSs exhibit higher peroxidase activity due to more catalytic active site exposure. This study provides a strategy to prepare an ultrathin 2D MOF with high catalytic activity and new insights for the construction of a biosensor to monitor AChE and new AD drugs.