Method for the determination of the concentration of an enzyme substrate and a sensor for carrying out the method
First Claim
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1. A method for the continuous and reversible determination of the concentration of an enzyme substrate in a specimen, comprising the steps of:
- (a) contacting said specimen containing said enzyme substrate with an enzyme specific for said substrate, selected from the group consisting of oxidases and oxygenases, to which a flavin coenzyme selected from the group consisting of flavin mononucleotide and flavin adenine dinucleotide is bonded, said flavin coenzyme converting to a reduced form by said enzyme substrate and to an oxidized form by molecular oxygen dissolved in said specimen,(b) measuring a change in intrinsic fluorescence of said flavin coenzyme under fluorescent excitation resulting from reduction of said flavin coenzyme occurring in step (a), and(c) determining the concentration of enzyme substrate in said specimen from the change in intrinsic fluorescence measured in step (b).
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Abstract
A method for the continuous and reversible determination of the concentration of an enzyme substrate such as glucose in an specimen, wherein the specimen is brought into contact with a corresponding enzyme selected from oxidases and oxygenases and to which a flavin coenzyme (FMN, FAD) is bonded, the flavin coenzyme changing to a reduced form by the enzyme substrate and to an oxidized form by molecular oxygen dissolved in the specimen, and the change in the fluorescence spectrum, produced by means of the reduction of the flavin coenzyme, is measured.
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8 Claims
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1. A method for the continuous and reversible determination of the concentration of an enzyme substrate in a specimen, comprising the steps of:
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(a) contacting said specimen containing said enzyme substrate with an enzyme specific for said substrate, selected from the group consisting of oxidases and oxygenases, to which a flavin coenzyme selected from the group consisting of flavin mononucleotide and flavin adenine dinucleotide is bonded, said flavin coenzyme converting to a reduced form by said enzyme substrate and to an oxidized form by molecular oxygen dissolved in said specimen, (b) measuring a change in intrinsic fluorescence of said flavin coenzyme under fluorescent excitation resulting from reduction of said flavin coenzyme occurring in step (a), and (c) determining the concentration of enzyme substrate in said specimen from the change in intrinsic fluorescence measured in step (b). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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Specification