Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive europium probe

被引：67

作者：

Wu, M ^{[1
]}

Lin, ZH ^{[1
]}

Schäferling, M ^{[1
]}

Dürkop, A ^{[1
]}

Wolfbeis, OS ^{[1
]}

机构：

[1] Univ Regensburg, Inst Analyt Chem Chemo & Biosensors, D-93040 Regensburg, Germany

来源：

ANALYTICAL BIOCHEMISTRY | 2005年 / 340卷 / 01期

关键词：

glucose oxidase; enzyme imaging; fluorescent probe; time-resolved imaging; enzyme activity assay;

D O I：

10.1016/j.ab.2005.01.050

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for optical imaging of the activity of glucose oxidase (GOx) using a fluorescent europium(III) tetracycline probe for hydrogen peroxide is presented. A decay time in the microsecond range and the large Stokes shift of 210 nm of the probe facilitate intensity-based, time-resolved, and decay-time-based imaging of glucose oxidase. Four methods for imaging the activity of GOx were compared, and rapid lifetime determination imaging was found to be the best in giving a linear range from 0.32 to 2.7 mUnit/mL. The detection limit is 0.32 mUnit/mL (1.7 ng mL(-1)) which is similar to that of the time-resolved (gated) imaging using a microtiterplate reader. Fluorescent imaging of the activity of GOx is considered to be a useful toot for GOx-based immunoassays with potential for high-throughput screening, immobilization studies, and biosensor array technologies. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：66 / 73

页数：8

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