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荧光法测定丁酰胆碱酯酶活性及其在生物成像和农药残留检测中的应用

Fluorescent Determination of Butyrylcholinesterase Activity and Its Application in Biological Imaging and Pesticide Residue Detection.

作者信息

Zhang Qian, Fu Caixia, Guo Xinjie, Gao Jian, Zhang Peng, Ding Caifeng

机构信息

Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE; Shandong Key Laboratory of Biochemical Analysis; Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong; College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, P.R. China.

出版信息

ACS Sens. 2021 Mar 26;6(3):1138-1146. doi: 10.1021/acssensors.0c02398. Epub 2021 Jan 27.

Abstract

Butyrylcholinesterase (BChE) is an essential human cholinesterase relevant to liver conditions and neurodegenerative diseases, which makes it a pivotal biomarker of health. It therefore remains challenging and highly desired to elaborate efficient chemical tools for BChE with simple operations and satisfactory working performance. In this work, a background-free detection strategy was built by virtue of the judicious coupling of a specific BChE-enzymatic reaction and in situ cyclization. High sensitivity with a low limit of detection (LOD) of 0.075 μg/mL could be readily achieved from the blank background and the as-produced emissive indicators, and the specific reaction site contributed to the high selectivity over other bio-species even acetylcholinesterase (AChE). In addition to the multifaceted spectral experiments to verify the sensing mechanism, this work assumed comprehensive studies on the application. The bio-investigation ranged from cells to an organism, declaring a noteworthy prospect in disease diagnosis, especially for Alzheimer's disease (AD), a common neurodegenerative disease with over-expressed BChE. Moreover, its excellent work for inhibition efficacy elucidation was also proved with the accuracy IC of tacrine for BChE (8.6 nM), giving rise to an expanded application for trace pesticide determination.

摘要

丁酰胆碱酯酶(BChE)是一种与肝脏疾病和神经退行性疾病相关的重要人体胆碱酯酶,这使其成为健康的关键生物标志物。因此,开发操作简单且性能良好的高效BChE化学工具仍然具有挑战性且备受期待。在这项工作中,通过将特定的BChE酶促反应与原位环化巧妙结合,构建了一种无背景检测策略。从空白背景和产生的发光指示剂中很容易实现低检测限(LOD)为0.075μg/mL的高灵敏度,并且特定的反应位点使得对其他生物物种甚至乙酰胆碱酯酶(AChE)具有高选择性。除了通过多方面的光谱实验验证传感机制外,这项工作还对其应用进行了全面研究。生物研究范围从细胞到生物体,在疾病诊断中展现出值得关注的前景,特别是对于阿尔茨海默病(AD)这种BChE过表达的常见神经退行性疾病。此外,它在阐明抑制效果方面的出色工作也通过他克林对BChE的准确半数抑制浓度(IC)(8.6 nM)得到证明,从而在痕量农药测定方面有了更广泛的应用。

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