Abstract
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.
| Language | English |
|---|---|
| Pages | 968-972 |
| Number of pages | 5 |
| Journal | Analyst |
| Volume | 136 |
| Issue number | 5 |
| Early online date | 17 Dec 2010 |
| DOIs | |
| Publication status | Published - 2011 |
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Keywords
- resonance-energy transfer
- in-vivo
- biosensor
- micocapsules
- assay
- TIC - Bionanotechnology
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A fluorescence lifetime-based fibre-optic glucose sensor using glucose/galactose-binding protein. / Saxl, Tania; Khan, Faaizah; Ferla, Matteo; Birch, David; Pickup, John.
In: Analyst, Vol. 136, No. 5, 2011, p. 968-972.Research output: Contribution to journal › Article
TY - JOUR
T1 - A fluorescence lifetime-based fibre-optic glucose sensor using glucose/galactose-binding protein
AU - Saxl, Tania
AU - Khan, Faaizah
AU - Ferla, Matteo
AU - Birch, David
AU - Pickup, John
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - resonance-energy transfer
KW - in-vivo
KW - biosensor
KW - micocapsules
KW - assay
KW - TIC - Bionanotechnology
U2 - 10.1039/c0an00430h
DO - 10.1039/c0an00430h
M3 - Article
VL - 136
SP - 968
EP - 972
JO - Analyst
T2 - Analyst
JF - Analyst
SN - 0003-2654
IS - 5
ER -