1 November 2005 Measurements of FRET in a Glucose-sensitive Affinity System with Frequency-domain Lifetime Spectroscopy
Feng Liang, Tianshu Pan, Eva M. Sevick-Muraca
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We report measurements of fluorescence resonance energy transfer (FRET) for glucose sensing in an established concanavalin A–dextran affinity system using frequency-domain lifetime spectroscopy. A dextran (MW 2 000 000) labeled with a small fluorescent donor molecule, Alexa Fluor 568, was used to competitively bind to a sugar-binding protein, concanavalin A, labeled with acceptor molecule, Alexa Fluor 647, in the presence of glucose. The FRET-quenching kinetics of the donor were analyzed from frequency-domain measurements as a function of both glucose and acceptor-protein concentrations using a Förster-type decay kinetics model. The results show that the frequency-domain measurements and donor decay kinetics can quantitatively indicate changes in the competitive binding of 0.09 μM dextran to labeled concanavalin A at a solution concentration of 10.67 μM in the presence of glucose at concentrations ranging from 0 to 224 mg/dL.

Feng Liang, Tianshu Pan, and Eva M. Sevick-Muraca "Measurements of FRET in a Glucose-sensitive Affinity System with Frequency-domain Lifetime Spectroscopy," Photochemistry and Photobiology 81(6), 1386-1394, (1 November 2005). https://doi.org/10.1562/2005-02-14-RA-440
Received: 14 February 2005; Accepted: 1 July 2005; Published: 1 November 2005

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