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

Shuo Li, Zhongyu Wei, Li Xiong, Qi Xu, Long Yu*, and Yuxiu Xiao*

Anal. Chem. 2022, 94, 49, 17263–17271

https://doi.org/10.1021/acs.analchem.2c04218

Abstract

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 (oPD) 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 oPD cascade polymerization strategy was proposed. That is, oPD was oxidized by Cu-BTC to oPD oligomers with strong yellow fluorescence, and oPD oligomers were further catalyzed to generate J-aggregation, which promotes the formation of oPD 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 oPD 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 oPD polymers and the fluorescence–photothermal dual-mode assay of enzyme activity.