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000850274 0247_ $$2doi$$a10.1021/acssensors.8b00143
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000850274 1001_ $$0P:(DE-Juel1)165927$$aHöfig, Henning$$b0
000850274 245__ $$aGenetically Encoded Förster Resonance Energy Transfer-Based Biosensors Studied on the Single-Molecule Level
000850274 260__ $$aWashington, DC$$bACS Publications$$c2018
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000850274 520__ $$aGenetically encoded Förster resonance energy transfer (FRET)-based biosensors for the quantification of ligand molecules change the magnitude of FRET between two fluorescent proteins upon binding a target metabolite. When highly sensitive sensors are being designed, extensive sensor optimization is essential. However, it is often difficult to verify the ideas of modifications made to a sensor during the sensor optimization process because of the limited information content of ensemble FRET measurements. In contrast, single-molecule detection provides detailed information and higher accuracy. Here, we investigated a set of glucose and crowding sensors on the single-molecule level. We report the first comprehensive single-molecule study of FRET-based biosensors with reasonable counting statistics and identify characteristics in the single-molecule FRET histograms that constitute fingerprints of sensor performance. Hence, our single-molecule approach extends the toolbox of methods aiming to understand and optimize the design of FRET-based biosensors.
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000850274 7001_ $$0P:(DE-Juel1)165764$$aOtten, Julia$$b1
000850274 7001_ $$0P:(DE-Juel1)145517$$aSteffen, Victoria$$b2
000850274 7001_ $$0P:(DE-Juel1)131522$$aPohl, Martina$$b3
000850274 7001_ $$0P:(DE-HGF)0$$aBoersma, Arnold J.$$b4
000850274 7001_ $$0P:(DE-HGF)0$$aFitter, Joerg$$b5$$eCorresponding author
000850274 773__ $$0PERI:(DE-600)2843497-3$$a10.1021/acssensors.8b00143$$gp. acssensors.8b00143$$n8$$p1462–1470$$tACS sensors$$v3$$x2379-3694$$y2018
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