Journal
ANALYTICAL BIOCHEMISTRY
Volume 340, Issue 1, Pages 66-73Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2005.01.050
Keywords
glucose oxidase; enzyme imaging; fluorescent probe; time-resolved imaging; enzyme activity assay
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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.
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