Fluorescence sensing of glucose

Leah Tolosa; Ignacy Gryczynski; Lisa Randers-Eichhorn; Jonathan D. Dattelbaum; Govind Rao; Joseph R. Lakowicz

doi:10.1117/12.347544

3 May 1999 Fluorescence sensing of glucose

Leah Tolosa, Ignacy Gryczynski, Lisa Randers-Eichhorn, Jonathan D. Dattelbaum, Govind Rao, Joseph R. Lakowicz

Proceedings Volume 3602, Advances in Fluorescence Sensing Technology IV; (1999) https://doi.org/10.1117/12.347544
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

We devised an optical assay for glucose based on the genetically-engineered glucose/galactose binding protein (GGBP) from E. coli and phase-modulation fluorometry. A single cysteine mutation was introduced at position 26 of GGBP. When labeled with the sulfhydryl-reactive probe I-ANS, GGBP showed a more than 50% decrease in florescence intensity with increasing glucose concentration (K_d approximately 1 (mu) M). This is consistent with the glucose-bound structure of GGBP where residue 26 becomes more exposed to the aqueous media. Since minimal lifetime changes were observed with glucose binding, a modulation sensor was devised wherein a long lifetime ruthenium metal-ligand complex (Ru) was painted on the surface of the cuvette containing ANS26-GGBP. Glucose binding resulted in changes in the relative intensities of ANS26-GGBP and Ru which were observed as dramatic changes in the modulation at a low frequency of 2.1 MHz. The modulation measured at 2.1 MHz accurately determines the glucose concentration to plus or minus 0.2 (mu) M.

Citation Download Citation

Leah Tolosa, Ignacy Gryczynski, Lisa Randers-Eichhorn, Jonathan D. Dattelbaum, Govind Rao, and Joseph R. Lakowicz "Fluorescence sensing of glucose", Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); https://doi.org/10.1117/12.347544

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