Characterization of multicomponent monosaccharide solutions using an enzyme-based sensor array

Theodore E. Curey, Adrian Goodey, Andrew Tsao, John Lavigne, Youngsoo Sohn, John T. McDevitt, Eric V. Anslyn, Dean Neikirk, Jason B. Shear

Research output: Contribution to journalArticlepeer-review


We report the development of a sensor for rapidly and simultaneously measuring multiple sugars in aqueous samples. In this strategy, enzyme-based assays are localized within an array of individually addressable sites on a micromachined silicon chip. Microspheres derivatized with monosaccharide-specific dehydrogenases are distributed to pyramidal cavities anisotropically etched in a wafer of silicon (100) and are exposed to sample solution that is forced through the cavities by a liquid chromatography pumping system. Production of fluorescent reporter molecules is monitored under stopped-flow conditions when localized dehydrogenase enzyme systems are exposed to their target sugars. We demonstrate the capability of this analysis strategy to quantify β-D-glucose and βD-galactose at low micromolar to millimolar levels, with no detectable cross-talk between assay sites. Analysis is achieved either through fluorescence detection of an initial dehydrogenase product (NADH, NADPH) or by production of a secondary fluorescent product created by hydride transfer from the reduced nicotinamide cofactor to a fluorogenic reagent. The array format of this sensor provides capabilities for redundant analysis of sugars and for monitoring levels of other solution components known to affect the activity of enzymes. The use of this strategy to normalize raw fluorescence signals is demonstrated by the determination of glucose and pH on a single chip. Alternatively, uncertainties in the activity of an immobilized enzyme can be accounted for using standard additions, an approach used here in the determination of serum glucose.

Original languageEnglish (US)
Pages (from-to)178-184
Number of pages7
JournalAnalytical Biochemistry
Issue number2
StatePublished - Jun 15 2001

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Characterization of multicomponent monosaccharide solutions using an enzyme-based sensor array'. Together they form a unique fingerprint.

Cite this