Method and system for determining the presence of adenosine triphosphate or flavin mononucleotide
First Claim
1. A reagent system for the determination of a coenzyme selected from the group consisting adenosine triphosphate and flavin mononucleotide in an aqueous liquid sample, comprising:
- (a) a coenzyme selected from adenine flavin mononucleotide triphosphate and flavin mononucleotide, one of which is the substance to be determined and the other of which is a component of the reagent system;
(b) an enzyme preparation having flavin adenine dinucleotide synthetase activity; and
(c) apo(glucose oxidase).
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Accused Products
Abstract
An enzymatic determination of one of the co-enzymes adenosine triphosphate (ATP) or flavin mono-nucleotide (FMN) based on the use of an flavine adenine dinucleotide (FAD) synthetase-active system in conjunction with the other coenzyme to generate FAD. The generated FAD can then be measured to determine the ATP or FMN concentration in the sample. Preferably, the generated FAD is measured by combination with a corresponding apoenzyme, preferably apo (glucose oxidase), and measuring the activity of the enzyme produced. The resultant active enzyme is preferably measured in a manner which results in a colorimetric response. The present invention provides an ATP or FMN assay which combines the sensitivity of bioluminescent assays with the convenience of a colorimetrically detectable response.
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Citations
12 Claims
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1. A reagent system for the determination of a coenzyme selected from the group consisting adenosine triphosphate and flavin mononucleotide in an aqueous liquid sample, comprising:
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(a) a coenzyme selected from adenine flavin mononucleotide triphosphate and flavin mononucleotide, one of which is the substance to be determined and the other of which is a component of the reagent system; (b) an enzyme preparation having flavin adenine dinucleotide synthetase activity; and (c) apo(glucose oxidase). - View Dependent Claims (2)
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3. An analytical method for determining adenosine triphosphate ("ATP") in picomole concentrations, said method comprising the steps of:
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combining a liquid sample with a reagent system, said system comprising in a single solution; flavin mononucleotide ("FMN"); flavin adenine dinucleotide ("FAD") synthetase; apo(glucose oxidase); a peroxidatively active substance; glucose; a metal cation; and a photogen, whereby said photogen provides a detectable optical response in the presence of hydrogen peroxide and the peroxidatively active substance, and detecting the optical response said response being an indication of the amount of ATP down to picomole concentrations in said liquid sample. - View Dependent Claims (4, 5, 6, 7)
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8. An analytical method for determining flavin mononucleotide ("FMN") in picomole concentrations, said method comprising the steps of:
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combining a liquid sample with a reagent system, said system comprising in a single solution; adenosine triphosphate ("ATP"); flavin adenine dinucleotide ("FAD") synthetase; apo(glucose oxidase); a peroxidatively active substance; glucose; a metal cation; and a photogen, whereby said photogen provides a detectable optical response in the presence of a hydrogen peroxide and the peroxidatively active substance, and detecting the optical response said response being an indication of the amount of FMN down to picomole concentrations in said liquid sample. - View Dependent Claims (9, 10)
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11. An analytical reagent system for determining adenosine triphosphate ("ATP") in picomole concentrations, said reagent system comprising:
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flavin mononucleotide ("FMN"); flavin adenine dinucleotide ("FAD") synthetase; apo(glucose oxidase); a peroxidatively active substance; glucose; a metal cation; and a photogen, whereby said photogen provides a detectable optical response in the presence of a hydrogen peroxide and the peroxidatively active substance.
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12. An analytical reagent system for determining flavin mononucleotide ("FMN") in picomole concentrations, said reagent system comprising:
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adenosine triphosphate ("ATP"); flavin adenine dinucleotide ("FAD") synthetase; apo(glucose oxidase); a peroxidatively active substance; glucose; a metal cation; and a photogen, whereby said photogen provides a detectable optical response in the presence of a hydrogen peroxide and the peroxidatively active substance.
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Specification