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| Naji, M. |
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| Motta, Antonella |
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| Aletan, Dirar |
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| Mohamed, Tarek |
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| Ertürk, Emre |
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| Taccardi, Nicola |
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| Kononenko, Denys |
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| Petrov, R. H. | Madrid |
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| Alshaaer, Mazen | Brussels |
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| Bih, L. |
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| Casati, R. |
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| Muller, Hermance |
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| Kočí, Jan | Prague |
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| Šuljagić, Marija |
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| Kalteremidou, Kalliopi-Artemi | Brussels |
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| Azam, Siraj |
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| Ospanova, Alyiya |
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| Blanpain, Bart |
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| Ali, M. A. |
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| Popa, V. |
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| Rančić, M. |
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| Ollier, Nadège |
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| Azevedo, Nuno Monteiro |
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| Landes, Michael |
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| Rignanese, Gian-Marco |
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Jenkins, Toby
University of Bath
in Cooperation with on an Cooperation-Score of 37%
Topics
Publications (8/8 displayed)
- 2018Development of an Infection-Responsive Fluorescent Sensor for the Early Detection of Urinary Catheter Blockagecitations
- 2016An in-situ infection detection sensor coating for urinary catheterscitations
- 2010Polymer-tethered bimolecular lipid membranescitations
- 2008Surface plasmon resonance-enhanced fluorescence implementation of a single-step competition assay: Demonstration of fatty acid measurement using an anti-fatty acid monoclonal antibody and a Cy5-labeled fatty acidcitations
- 2007Adhesion improvement of plasma-polymerized maleic anhydride films on gold using HMDSO/O-2 adhesion layerscitations
- 2007The effect of the ionophore valinomycin on biomimetic solid supported lipid DPPTE/EPC membranescitations
- 2002Chemical structure and properties of plasma-polymerized maleic anhydride filmscitations
- 2000Pulsed plasma deposited maleic anhydride thin films as supports for lipid bilayerscitations
Places of action
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article
Surface plasmon resonance-enhanced fluorescence implementation of a single-step competition assay: Demonstration of fatty acid measurement using an anti-fatty acid monoclonal antibody and a Cy5-labeled fatty acid
Abstract
The development of a single-step, separation-free method for measurement of low concentrations of fatty acid using a surface plasmon resonance-enhanced fluorescence competition assay with a surface-bound antibody is described. The assay behavior was unexpectedly complex. A nonlinear coverage-dependent self-quenching of emission from surface-bound fluorescent label was deduced from the response kinetics and attributed to a surface plasmon-mediated energy transfer between adsorbed fluorophores, modified by the effects of plasmon interference. Principles of assay design to avoid complications from such effects are discussed. An anti-fatty acid mouse monoclonal antibody reacting to the alkyl chain was prepared and supported on a gold chip at a spacing appropriate for surface-plasmon field-enhanced fluorescence spectroscopy (SPEFS), by applying successively a self-assembled biotinylated monolayer, then streptavidin, then biotinylated protein A, and then the antibody, which was crosslinked to the protein A. Synthesis of a fluorescently (Cy5) tagged C-11 fatty acid is reported. SPEFS was used to follow the kinetics of the binding of the labeled fatty acid to the antibody, and to implement a competition assay with free fatty acid (undecanoic acid), sensitive at the 1 mu M scale, a sensitivity limit caused by the low affinity of antibodies for free fatty acids, rather than the SPEFS technique itself. Free fatty acid concentration in human serum is in the range 0.1-1 mM, suggesting that this measurement approach could be applied in a clinical diagnostic context. Finally, a predictive, theoretical model of fatty acid binding was developed that accounted for the observed "overshoot" kinetics. (c) 2008 Elsevier Inc. All rights reserved.