Glucose sensor for low-cost lifetime-based sensing using a genetically engineered protein

Leah Tolosa, Ignacy Gryczynski, Lisa R. Eichhorn, Jonathan D. Dattelbaum, Felix N. Castellano, Govind Rao, Joseph R. Lakowicz

Research output: Contribution to journalArticlepeer-review

167 Scopus citations

Abstract

We describe a glucose sensor based on a mutant glucose/galactose binding protein (GGBP) and phase-modulation fluorometry. The GGBP from Escherichia coli was mutated to contain a single cysteine residue at position 26. When labeled with a sulfhydryl-reactive probe 2-(4'- iodoacetamidoanilino)naphthalene-6-sulfonic acid, the labeled protein displayed a twofold decrease in intensity in response to glucose, with a dissociation constant near 1 μM glucose. The ANS-labeled protein displayed only a modest change in lifetime, precluding lifetime-based sensing of glucose. A modulation sensor was created by combining ANS26-GGBP with a long- lifetime ruthenium (Ru) metal-ligand complex on the surface of the cuvette. Binding of glucose changed the relative intensity of ANS26-GGBP and the Ru complex, resulting in a dramatic change in modulation at a low frequency of 2.1 MHz. Modulation measurements at 2.1 MHz were shown to accurately determine the glucose concentration. These results suggest an approach to glucose sensing with simple devices.

Original languageEnglish
Pages (from-to)114-120
Number of pages7
JournalAnalytical Biochemistry
Volume267
Issue number1
DOIs
StatePublished - 1 Feb 1999

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