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August 09, 2009
Functional Polymers: Fluorescent Enzyme-Coupled Approach for Enzyme Detection Using Conjugated Polymers
Fude Feng, Libing Liu,* and Shu Wang*
Sensitive enzyme detections based on conjugated polymers (CPs) is of great interest due to its importance in clinical diagnosis, drug discovery and enzyme engineering. Shu Wang and co-workers present a simple, rapid, continuous and homogeneous fluorescent assay for β-galactosidase which combines an enzyme-coupled reaction and the signal amplification property of 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 the conversion to quinone (secondary product) with fluorescence quenching ability by the tyrosinase. The fluorescence of PFP-SO3- is efficiently quenched by the quinone via an electron transfer process. The detection limit of this assay is less than 0.0005U x mL-1, which is better than that of the electrochemical method and comparable to that of the most sensitive chemiluminescent techniques. In principle, this sensor mechanism will extend the application window of CPs for wide-spectrum enzyme detections and could be expanded to a high-throughput manner.
Macromol. Chem. Phys., DOI: 10.1002/macp.200900264
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