Water-Soluble Conjugated Polyelectrolyte-Based Fluorescence Enzyme Coupling Protocol for Continuous and Sensitive β-Galactosidase Detection
✍ Scribed by Fude Feng; Libing Liu; Shu Wang
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 331 KB
- Volume
- 210
- Category
- Article
- ISSN
- 1022-1352
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✦ Synopsis
Abstract
A simple, rapid, continuous and homogeneous fluorescent assay for β‐galactosidase was developed, combining an enzyme‐coupled reaction and signal amplification property of conjugated polyelectrolytes (CPs). The procedure is based on a sequence of two coupled biocatalytic steps in which the β‐galactosidase hydrolyzes its substrate to a phenol derivative followed by conversion to quinone (secondary product) with fluorescence quenching ability by the tyrosinase. The fluorescence of PFP−SO is efficiently quenched by the quinone via an electron transfer process. The limit of detection (LOD) of this assay is less than 0.0005 U · mL^−1^, which is better than that of electrochemical method and is comparable to that of most sensitive chemiluminescent techniques. In principle, this sensor mechanism will extend the application window of CPs for wide‐spectrum enzyme detections. This “mix‐and‐detect” approach could be expanded to a high‐throughput manner.
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