Enzyme-Nanozyme Cascade Flow Reactor Synergy with Deep Learning for Differentiation and Point-of-Care Testing of Multiple Organophosphorus Pesticides

Abstract

Combining nanotechnology with biocatalysts, the construction of a cascade continuous flow reactor is a cutting-edge strategy to enhance the stability and efficiency of catalysis. In this study, C60@MOF-545-Fe nanozyme is synthesized by utilizing fullerene (C60) as a guest encapsulated inside a metal-organic framework (MOF-545-Fe), the unique host-guest interaction optimizes the oxidase (OXD)- and peroxidase (POD)-like activities, based on which a cascading catalytic strategy is proposed without external energy input. Simultaneously, C60@MOF-545-Fe offers the potential to effectively flow at the nanoscale through its unique nanostructure cavity and spatially confined environment. Therefore, the AChE/C60@MOF-545-Fe enzyme-nanozyme continuous flow reactor is constructed by combining C60@MOF-545-Fe with acetylcholinesterase (AChE) through supramolecular interactions. Notably, the reactor not only achieves the simultaneous detection of glyphosate, omethoate, and paraoxon but also efficiently differentiates these three organophosphorus pesticides (OPs) by applying the differences in the responses of the three array channels. Subsequently, a portable platform is developed utilizing the YOLO v5-OPs model based on deep learning, enabling the direct output fitting equation through terminals to achieve rapid point-of-care testing (POCT) of OPs. This work not only provides a promising strategy for hazard detection systems but also opens up new avenues for the design of technologies based on flow reactors.