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Alternative, non-electrochemistry-based technologies for continuous glucose monitoring are needed for eventual use in diabetes mellitus. As part of a programme investigating fluorescent glucose sensors, we have developed fibre-optic biosensors using glucose/galactose binding protein (GBP) labelled with the environmentally sensitive fluorophore, Badan. GBP-Badan was attached via an oligohistidine-tag to the surface of Ni-nitrilotriacetic acid (NTA)-functionalized agarose or polystyrene beads. Fluorescence lifetime increased in response to glucose, observed by fluorescence lifetime imaging microscopy of the GBP-Badan-beads. Either GBP-Badan agarose or polystyrene beads were loaded into a porous chamber at the end of a multimode optical fibre. Fluorescence lifetime responses were recorded using pulsed laser excitation, high speed photodiode detection and time-correlated single-photon counting. The maximal response was at 100 mM glucose with an apparent K(d) of 13 mM (agarose) and 20 mM (polystyrene), and good working-day stability was demonstrated. We conclude that fluorescence lifetime fibre-optic glucose sensors based on GBP-Badan are suitable for development as clinical glucose monitors.

Original publication

DOI

10.1039/c0an00430h

Type

Journal article

Journal

The Analyst

Publication Date

03/2011

Volume

136

Pages

968 - 972

Addresses

Diabetes Research Group, King's College London School of Medicine, Guy's Hospital, London, SE1 1UL, UK.

Keywords

Nitrilotriacetic Acid, Polystyrenes, Glucose, Sepharose, Periplasmic Binding Proteins, Calcium-Binding Proteins, Monosaccharide Transport Proteins, Microscopy, Fluorescence, Biosensing Techniques, Microspheres, Fluorescence, Time Factors, Optical Fibers