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A Switch‐On Fluorescence Assay for Bacterial β‐Lactamases with Amyloid Fibrils as Fluorescence Enhancer and Visual Tool
Author(s) -
Zou Lan,
Cheong WingLam,
Chung WaiHong,
Leung YunChung,
Wong KwokYin,
Wong ManKin,
Chan PakHo
Publication year - 2010
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201001633
Subject(s) - fluorescence , fluorophore , chemistry , lysozyme , fibril , biophysics , biochemistry , biology , optics , physics
Herein is described the development of a novel switch‐on fluorescence assay for detecting β‐lactamases. The fluorescence assay comprises two components: solid beads coated with a β‐lactam antibiotic, which is linked to an environment‐sensitive fluorophore (dansylaminothiophenol, DTA), and amyloid fibrils of hen lysozyme (acting as fluorescence enhancer and visual tool). In the presence of the clinically significant TEM‐1 β‐lactamase, the DTA–antibiotic complex on the solid beads is hydrolyzed, thus releasing the DTA dye into solution. The DTA dye is only weakly fluorescent in solution but gives strong green fluorescence upon binding to lysozyme fibrils. These strongly fluorescent DTA‐bound fibrils can be easily visualized by the naked eye upon illumination of the sample with a simple UV lamp. The fluorescence assay can detect TEM‐1 at low concentration (0.01 n M ). In contrast, no observable fluorescence appears when the fluorescence assay is performed on samples without the TEM‐1 β‐lactamase.