Elsevier

Analytical Biochemistry

Volume 267, Issue 1, 1 February 1999, Pages 114-120
Analytical Biochemistry

Regular Article
Glucose Sensor for Low-Cost Lifetime-Based Sensing Using a Genetically Engineered Protein,☆☆

https://doi.org/10.1006/abio.1998.2974Get rights and content

Abstract

We describe a glucose sensor based on a mutant glucose/galactose binding protein (GGBP) and phase-modulation fluorometry. The GGBP fromEscherichia coliwas 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.

References (44)

  • K. Yokoyama et al.

    Anal. Chim. Acta

    (1989)
  • G.M. Schier et al.

    Diabetes Res. Clin. Pract.

    (1988)
  • D. Meadows et al.

    Talanta

    (1988)
  • J.R. Lakowicz et al.

    Anal. Chim. Acta

    (1993)
  • L. Tolosa et al.

    Anal. Biochem.

    (1997)
  • L. Tolosa et al.

    Sensors Actuators B

    (1997)
  • S.L. Jacques et al.

    J. Invest. Dermatol.

    (1987)
  • W. Boos et al.

    J. Biol. Chem.

    (1972)
  • W. Boos

    J. Biol. Chem.

    (1972)
  • J.R. Lakowicz et al.

    Biophys. Chem.

    (1985)
  • X-Q. Guo et al.

    Anal. Biochem.

    (1997)
  • E. Terpetschnig et al.

    Methods Enzymol.

    (1997)
  • W. Boos et al.

    J. Biol. Chem.

    (1971)
  • J. Slavik

    Biochim. Biophy. Acta

    (1982)
  • K.W. Berndt et al.

    Anal. Biochem.

    (1992)
  • H.M. Heise et al.

    Artif. Organs

    (1994)
  • M.R. Robinson et al.

    Clin. Chem.

    (1992)
  • J.J. Burmeister et al.

    Photochem. Photobiol.

    (1998)
  • H.M. Heise et al.

    Artif. Organs

    (1994)
  • W.F. March et al.

    Trans. Am. Soc. Artif. Intern. Organ

    (1982)
  • B. Rabinovitch et al.

    Diabetes Care

    (1982)
  • Cited by (0)

    Dedicated to Dr. Henryk Szmacinski on the occasion of his 50th birthday.

    ☆☆

    Lakowicz, J. R.

    2

    To whom correspondence should be addressed.

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