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基于蛋白-无机杂化纳米花-琼脂糖水凝胶平台的即时检测乙酰胆碱的方法。

Protein-Inorganic Hybrid Nanoflower-Rooted Agarose Hydrogel Platform for Point-of-Care Detection of Acetylcholine.

机构信息

State Key Laboratory on Integrated Optoelectronics, College of Electronic Science and Engineering , Jilin University , Changchun 130012 , People's Republic of China.

School of Mechanical and Materials Engineering , Washington State University , Pullman , Washington 99164 , United States.

出版信息

ACS Appl Mater Interfaces. 2019 Mar 27;11(12):11857-11864. doi: 10.1021/acsami.8b21571. Epub 2019 Mar 14.

Abstract

Rapid and precise profiling of acetylcholine (ACh) has become important for diagnosing diseases and safeguarding health care because of its pivotal role in the central nervous system. Herein, we developed a new colorimetric sensor based on protein-inorganic hybrid nanoflowers as artificial peroxidase, comprising a test kit and a smartphone reader, which sensitively quantifies ACh in human serum. In this sensor, ACh indirectly triggered the substrate reaction with the help of a multienzyme system including acetylcholinesterase, choline oxidase, and mimic peroxidase (nanoflowers), accompanying the enhancement of absorbance intensity at 652 nm. Therefore, the multienzyme platform can be used to detect ACh via monitoring the change of the absorbance in a range from 0.0005 to 6.0 mmol L. It is worth mentioning that the platform was used to prepare a portable agarose gel-based kit for rapid qualitative monitoring of ACh. Coupling with ImageJ program, the image information of test kits can be transduced into the hue parameter, which provides a directly quantitative tool to identify ACh. Based on the advantages of simple operation, good selectivity, and low cost, the availability of a portable kit for point-of-care testing will achieve the needs of frequent screening and diagnostic tracking.

摘要

由于乙酰胆碱(ACh)在中枢神经系统中起着关键作用,因此快速准确地分析其结构对于疾病诊断和保健变得尤为重要。在此,我们开发了一种基于蛋白质-无机杂化纳米花的新型比色传感器作为人工过氧化物酶,包括试剂盒和智能手机读取器,可灵敏地定量人血清中的 ACh。在该传感器中,ACh 在包括乙酰胆碱酯酶、胆碱氧化酶和模拟过氧化物酶(纳米花)在内的多酶体系的帮助下间接触发底物反应,伴随着 652nm 处吸光度强度的增强。因此,该多酶平台可通过监测从 0.0005 到 6.0mmol/L 范围内吸光度的变化来检测 ACh。值得一提的是,该平台用于制备一种基于琼脂糖凝胶的便携式试剂盒,用于快速定性监测 ACh。与 ImageJ 程序耦合后,试剂盒的图像信息可以转化为色调参数,为识别 ACh 提供了一种直接定量工具。基于操作简单、选择性好、成本低的优点,该便携式即时检测试剂盒的可用性将满足频繁筛查和诊断跟踪的需求。

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