Time-Resolved Levodopa Cascade Polymerization Tuned by Bimetallic MOF Fluorescent Nanozyme and Boric Acid for Butyrylcholinesterase Activity Dual-Mode Assay

Abstract

A ratiometric fluorescence-photothermal dual-mode assay method is constructed for the detection of butyrylcholinesterase (BChE) activity based on time-resolved levodopa (L-DOPA) cascade polymerization. First, a newly designed bimetallic metal–organic framework (MOF), Eu/Co-DPA (DPA: pyridine-2,6-dicarboxylic acid), is screened out as a fluorescent nanozyme with high catalytic activity and superior luminescence properties. In the presence of boric acid (BA), L-DOPA forms BA-esterified L-DOPA, which is catalyzed by Eu/Co-DPA to form the oligomers with strong blue fluorescence. Meanwhile, the red fluorescence of Eu/Co-DPA is quenched by the oligomers, generating a sensitive turn-on/off ratiometric fluorescence response. As polymerization time increases, Eu/Co-DPA cleaves the borate ester bonds to expose the catechol structures of the oligomers, which facilitates the further oxidation and polymerization of the oligomers, promoting the formation of poly(L-DOPA) nanoparticles with a high photothermal conversion efficiency (30.33%). Then, by using thiocholine (butyrylthiocholine enzymolysis product) to inhibit the catalytic activity of Eu/Co-DPA, BChE activity is detected through the change in fluorescence and photothermal dual signals. Both assay modes have low detection limits (0.021 and 0.024 U L –1 ) and high accuracy (93.3–105.3% recovery). The detection results of real human serum indicate that both assay modes show 100.0% agreement with the standard method. To our knowledge, this work first combines bimetallic MOFs and a BA regulator to tune the structure of L-DOPA polymers, providing a pathbreaking paradigm for preparing catecholamine-based fluorescence-photothermal organic polymers.