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

共 50 条

[41]

TSUJI A, 1987, J STEROID BIOCHEM, V27, P33

[42] Glucose/O2 biofuel cell operating at physiological conditions [J].

Tsujimura, S ;

Kano, K ;

Ikeda, T .

ELECTROCHEMISTRY, 2002, 70 (12) :940-942

[43] Two-photon excitation fluorometric measurement of homogeneous microparticle immunoassay for C-reactive protein [J].

Waris, ME ;

Meltola, NJ ;

Soini, JT ;

Soini, E ;

Peltola, OJ ;

Hänninen, PE .

ANALYTICAL BIOCHEMISTRY, 2002, 309 (01) :67-74

[44]

Wolfbeis OS, 2002, ANGEW CHEM INT EDIT, V41, P4495, DOI 10.1002/1521-3773(20021202)41:23<4495::AID-ANIE4495>3.0.CO

[45]

2-I

[46] Reversible optical sensor membrane for hydrogen peroxide using an immobilized fluorescent probe, and its application to a glucose biosensor [J].

Wolfbeis, OS ;

Schäferling, M ;

Dürkop, A .

MICROCHIMICA ACTA, 2003, 143 (04) :221-227

[47] TRANSIENT DIGITIZER FOR THE DETERMINATION OF MICROSECOND LUMINESCENCE LIFETIMES [J].

WOODS, RJ ;

SCYPINSKI, S ;

LOVE, LJC ;

ASHWORTH, HA .

ANALYTICAL CHEMISTRY, 1984, 56 (08) :1395-1400

[48] Time-resolved enzymatic determination of glucose using a fluorescent europium probe for hydrogen peroxide [J].

Wu, M ;

Lin, ZH ;

Dürkop, A ;

Wolfbeis, OS .

ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2004, 380 (04) :619-626

[49] Plugging into enzymes: Nanowiring of redox enzymes by a gold nanoparticle [J].

Xiao, Y ;

Patolsky, F ;

Katz, E ;

Hainfeld, JF ;

Willner, I .

SCIENCE, 2003, 299 (5614) :1877-1881

[50] An SECM detection scheme with improved sensitivity and lateral resolution: Detection of galactosidase activity with signal amplification by glucose dehydrogenase [J].

Zhao, C ;

Wittstock, G .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2004, 43 (32) :4170-4172

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