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Water‐Soluble Conjugated Polyelectrolyte‐Based Fluorescence Enzyme Coupling Protocol for Continuous and Sensitive β ‐Galactosidase Detection
Author(s) -
Feng Fude,
Liu Libing,
Wang Shu
Publication year - 2009
Publication title -
macromolecular chemistry and physics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.57
H-Index - 112
eISSN - 1521-3935
pISSN - 1022-1352
DOI - 10.1002/macp.200900264
Subject(s) - chemistry , fluorescence , combinatorial chemistry , detection limit , polyelectrolyte , substrate (aquarium) , quenching (fluorescence) , conjugated system , quinone , biosensor , electron transfer , photochemistry , chromatography , organic chemistry , biochemistry , polymer , physics , oceanography , quantum mechanics , geology
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 3 −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.