基于氧化还原反应界面可视化定量检测辣根过氧化物酶

引用本文: 孔昊, 张强, 张薇, 刘伟文, 曹成喜, 樊柳荫. 基于氧化还原反应界面可视化定量检测辣根过氧化物酶[J]. 色谱, 2020, 38(2): 177-182. doi: 10.3724/SP.J.1123.2019.04030 shu

Citation: KONG Hao, ZHANG Qiang, ZHANG Wei, LIU Weiwen, CAO Chengxi, FAN Liuyin. Visualized and quantitative detection of horseradish peroxidase based on redox reaction boundary[J]. Chinese Journal of Chromatography, 2020, 38(2): 177-182. doi: 10.3724/SP.J.1123.2019.04030 shu

基于氧化还原反应界面可视化定量检测辣根过氧化物酶

孔昊 ^1,2, ,
张强 ^2,3, ,
张薇 ^2, ,
刘伟文 ^3, ,
曹成喜 ^3, ,
樊柳荫 ^1,

1.
上海交通大学学生创新中心, 上海 200240;
2.
上海交通大学生命科学技术学院, 上海 200240;
3.
上海交通大学电子信息与电气工程学院, 上海 200240
基金项目:
国家自然科学基金项目（21675067，31727801，21605101）；上海市科学技术委员会项目（15142200300）.

摘要: 该文建立了一种可视化的、基于氧化还原反应界面移动距离定量检测辣根过氧化物酶（HRP）的方法。比较了隐色结晶紫显色体系和3，3'，5，5'-四甲基联苯胺显色体系对HRP的显色效率，并构建了基于3，3'，5，5'-四甲基联苯胺显色体系的氧化还原反应电泳滴定模型。同时，文中还设计了适用于该模型的小型化、便携式滴定检测芯片，并对滴定通道凝胶中组分进行了优化。结果表明，界面移动距离与HRP浓度存在对数线性关系，检测灵敏度可达0.002 mg/L，且可在10 min内完成HRP的裸眼检测。该方法不需要配备信号读取装置，用户只需要读取有色界面移动的距离即可实现对待测物的可视化定量检测，对于即时检测具有潜在的应用价值。

关键词:

English

Visualized and quantitative detection of horseradish peroxidase based on redox reaction boundary

1.
Student Innovation Center, Shanghai Jiao Tong University, Shanghai 200240, China;
2.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;
3.
School of Electronic Information & Electrical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
Received Date: 16 April 2019
Fund Project:
National Natural Science Foundation of China (Nos. 21675067, 31727801, 21605101)

Abstract: A new portable method for the detection of horseradish peroxidase (HRP) is reported in this paper. Visualized and quantitative detection of HRP was achieved based on the theory of electrophoresis titration (ET) and redox reaction boundary (RB). The ET-RB model was built on the 3,3',5,5'-tetramethyl benzidine chromogenic system because it significantly increases the efficiency of the chromogenic reaction and can react with L-ascorbic acid. A series of experiments were performed on a tiny and portable titration chip based on the material of polymethyl methacrylate designed for the developed model. The composition of the titration gel was optimized to ensure higher detection sensitivity and stable detection performance. The experimental results manifested a log-linear relationship between the moving distance of the RB and the HRP concentration. The dynamic range of the developed method ranged from 0.002 to 0.073 mg/L, and the detection could be accomplished within 10 min. Visualized and quantitative detection could be realized by reading the moving distance of the colored boundary with the naked eyes instead of using any signal-reading device or analyzing instrument. Thus, this method has great potential for further application to the development of a real-time detection method for various target substances based on the detection of peroxidase activity.

Key words:

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基于氧化还原反应界面可视化定量检测辣根过氧化物酶

English

Visualized and quantitative detection of horseradish peroxidase based on redox reaction boundary

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