Fluorescence intensity- and lifetime-based glucose sensing using an engineered high-K(d) mutant of glucose/galactose-binding protein

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Abstract

We synthesized mutants of glucose/galactose-binding protein (GBP), labeled with the environmentally sensitive fluorophore Badan, with the aim of producing a fluorescence-based glucose sensing system with an operating range compatible with continuous glucose monitoring in patients with diabetes mellitus. From five mutants tested, the triple mutant H152C/A213R/L238S-Badan showed a large (200%) maximal increase in fluorescence intensity on the addition of glucose, with a binding constant (K-d) of 11 mM, an operating range of approximately 1-100 mM, and similar responses in buffer and serum. The mean fluorescence lifetime of this mutant also increased by 70% on the addition of glucose. We conclude that the GBP mutant H152C/A213R/L238S, when labeled with Badan, is suitable for development as a robust sensor for in vivo glucose monitoring in diabetes.
Original languageEnglish
Pages (from-to)39 - 43
Number of pages5
JournalAnalytical Biochemistry
Volume399
Issue number1
DOIs
Publication statusPublished - 1 Apr 2010

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