A low perfusion rate microreactor for the continous monitoring of enzyme characteristics: applications for glucose oxidase.

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Abstract

This report describes a versatile and robust microreactor for bioactive proteins physically immobilized on a polyether sulfone filter. The potential of the reactor is illustrated with glucose oxidase immobilized on a filter with a cut-off value of 30 kDa. A flow-injection system was used to deliver the reactants and the device was linked on-line to an electrochemical detector. The microreactor was used for on-line preparation of apoglucose oxidase in strong acid and its subsequent reactivation with flavin adenine dinucleotide. In addition we describe a miniaturized version of the microreactor used to assess several characteristics of femtomole to attomole amounts of glucose oxidase. A low negative potential over the electrodes was used when ferrocene was the mediator in combination with horseradish peroxidase, ensuring the absence of oxidation of electro-active compounds in biological fluids. A low backpressure at very low flow rates is an advantage, which increases the sensitivity. A variety of further applications of the microreactor are suggested.
Original languageEnglish
Pages (from-to)2029-2033
JournalAnalytical and Bioanalytical Chemistry
Volume389
Issue number6
DOIs
Publication statusPublished - 2007

Keywords

  • glyceraldehyde-3-phosphate dehydrogenase
  • reactor
  • reconstitution
  • spectrometry

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