Methods for determination of enzyme reactions
First Claim
1. A method for determining the activity of an enzyme in a liquid sample treated with a coenzyme and a substrate for a reaction involving a change in the coenzyme from a first form to a second form, utilizing a three-electrode potentiostat including an auxiliary electrode, comprising the steps of:
- electro-sensitively contacting said treated sample simultaneously with the electrodes of said potentiostat, applying a voltage to said auxiliary electrode to support said reaction, modulating said voltage to change said coenzyme alternately between said first and second forms, and measuring at least one of said forms during said reaction.
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Accused Products
Abstract
A method, for microcoulometric determination either of the activity of an enzyme in a liquid sample or the concentration of a substrate for a reaction involving a change in a coenzyme from a first form thereof to a second form. At intervals during the reaction, both coenzyme forms are measured essentially simultaneously utilizing an electrolytic cell, to indicate either such enzyme activity or substrate concentration, depending upon which is the unknown factor.
18 Citations
11 Claims
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1. A method for determining the activity of an enzyme in a liquid sample treated with a coenzyme and a substrate for a reaction involving a change in the coenzyme from a first form to a second form, utilizing a three-electrode potentiostat including an auxiliary electrode, comprising the steps of:
- electro-sensitively contacting said treated sample simultaneously with the electrodes of said potentiostat, applying a voltage to said auxiliary electrode to support said reaction, modulating said voltage to change said coenzyme alternately between said first and second forms, and measuring at least one of said forms during said reaction.
- View Dependent Claims (2, 3)
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4. A method for determining the activity of an enzyme in a liquid sample treated with a coenzyme and a substrate for a reaction involving a change in the coenzyme from a first form to a second form, utilizing a three-electrode potentiostat including an auxiliary electrode, comprising the steps of:
- electrosensitively contacting said treated sample simultaneously with the electrodes of said potentiostat, pulsing said auxiliary electrode over a period of time with voltage pulses so as to be biased above and below a baseline voltage, and measuring over a period of time the depletion of said first coenzyme form during said reaction.
- View Dependent Claims (5, 6, 7, 8, 9, 10)
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11. A method for determining the concentration of a substrate in a liquid sample treated with an enzyme and a coenzyme for a reaction involving a change in the coenzyme from a first form to a second form, utilizing a three-electrode potentiostat including an auxiliary electrode, comprising the steps of:
- electrosensitively contacting said treated sample with the electrodes of said potentiostat simultaneously, applying a first voltage to said auxiliary electrode over a period of time to change said coenzyme to said second form, applying a modulating voltage to said first voltage, so as to eliminate interference by other reactants during measurement of said second form, and measuring over a period of time said second form.
Specification