In Situ Formation of o -Phenylenediamine Cascade Polymers Mediated by Metal-Organic Framework Nanozymes for Fluorescent and Photothermal Dual-Mode Assay of Acetylcholinesterase Activity.

A fluorescent and photothermal dual-mode assay method was established for the detection of acetylcholinesterase (AChE) activity based on in situ formation of o -phenylenediamine ( o PD) cascade polymers. First, copper metal-organic frameworks of benzenetricarboxylic acid (Cu-BTC) were screened out as nanozymes with excellent oxidase-like activity and confinement catalysis effect. Then, an ingenious o PD cascade polymerization strategy was proposed. That is, o PD was oxidized by Cu-BTC to o PD oligomers with strong yellow fluorescence, and o PD oligomers were further catalyzed to generate J-aggregation, which promotes the formation of o PD polymer nanoparticles with a high photothermal effect. By utilizing thiocholine (enzymolysis product of acetylthiocholine) to inhibit the Cu-BTC catalytic effect, AChE activity was detected through the fluorescence-photothermal dual-signal change of o PD oligomers and polymer nanoparticles. Both assay modes have low detection limitation (0.03 U L -1 for fluorescence and 0.05 U L -1 for photothermal) and can accurately detect the AChE activity of human serum (recovery 85.0-111.3%). The detection results of real serum samples by fluorescent and photothermal dual modes are consistent with each other (relative error ≤ 5.2%). It is worth emphasizing that this is the first time to report the high photothermal effect of o PD polymers and the fluorescence-photothermal dual-mode assay of enzyme activity.