Ultrasensitivity Detecting AChE through “Covalent Assembly” and Signal Amplification Strategic Approaches and Applied to Screen Its Inhibitor

An ultrasensitivity detecting assay for acetylcholinesterase (AChE) activity was developed based on “covalent assembly” and signal amplification strategic approaches. After hydrolyzing thioacetylcholine by AChE and participation of thiol in a self-inducing cascade accelerated by the Meldrum acid der...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-03, Vol.95 (9), p.4503-4512
Main Authors: Zhao, Yongwei, Shen, Ao, Hao, Xiaohui, Li, Mengwen, Hou, Lala, Li, Ziqi, Duan, Ruochen, Du, Man, Li, Xue, Wang, Xuebing, Zhao, Xiuqing, Yang, Yunxu
Format: Article
Language:English
Subjects:
Acetylcholinesterase
Amplification
Assembly
Chemistry
Cholinesterase Inhibitors - pharmacology
Fluorescence
Humans
Hydrogels
Sulfhydryl Compounds
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Summary:An ultrasensitivity detecting assay for acetylcholinesterase (AChE) activity was developed based on “covalent assembly” and signal amplification strategic approaches. After hydrolyzing thioacetylcholine by AChE and participation of thiol in a self-inducing cascade accelerated by the Meldrum acid derivatives of 2-[bis­(methylthio) methylene] malonitrile (CA-2), mercaptans triggered an intramolecular cyclization assembly by the probe of 2-(2,2-dicyanovinyl)-5-(diethylamino) phenyl 2,4-dinitrobenzenesulfonate (Sd-I) to produce strong fluorescence. The limit of detection for AChE activity was as low as 0.0048 mU/mL. The detection system also had a good detecting effect on AChE activity in human serum and could also be used to screen its inhibitors. By constructing a Sd-I@agarose hydrogel with a smartphone, a point-of-care detection of AChE activity was achieved again.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.2c05313