Determination of substrate of coenzyme requiring enzyme
First Claim
1. A method of assaying within a liquid sample a substrate for a coenzyme-requiring enzyme which method comprises contacting the sample with an assay system including (i) a substrate analogue and (ii) a moiety which specifically binds the substrate without catalyzing the conversion of the substrate;
- the substrate analogue being a substance which competes with the substrate for binding to the moiety, and determining the presence or amount of the substrate by monitoring the extent to which there are formed or remain complexes between the moiety and at least one of the substrate analogue and the substrate;
wherein the moiety is selected from the group consisting of an apoenzyme which is not bound to its functional coenzyme, a partly denatured enzyme, a deactivated enzyme, a genetically engineered deactivated enzyme and a deactivated artificial enzyme.
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Abstract
The invention relates to a competitive binding assay technique for substances (e.g. small molecules having no antigenic determinant) which can serve as substrates for coenzyme-requiring enzymes. The technique involves contacting a liquid sample with an assay system including (i) a substrate analogue and (ii) a moiety capable of binding specifically to the substrate without catalyzing the conversion of substrate to product. The substrate analogue is a substance capable of binding to said moiety competitively with any of the substrate present in the sample. The presence or amount of the substrate is determined by monitoring the extent to which there are formed or remain complexes between the moiety and substrate or substrate analogue.
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Citations
10 Claims
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1. A method of assaying within a liquid sample a substrate for a coenzyme-requiring enzyme which method comprises contacting the sample with an assay system including (i) a substrate analogue and (ii) a moiety which specifically binds the substrate without catalyzing the conversion of the substrate;
- the substrate analogue being a substance which competes with the substrate for binding to the moiety, and determining the presence or amount of the substrate by monitoring the extent to which there are formed or remain complexes between the moiety and at least one of the substrate analogue and the substrate;
wherein the moiety is selected from the group consisting of an apoenzyme which is not bound to its functional coenzyme, a partly denatured enzyme, a deactivated enzyme, a genetically engineered deactivated enzyme and a deactivated artificial enzyme. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
- the substrate analogue being a substance which competes with the substrate for binding to the moiety, and determining the presence or amount of the substrate by monitoring the extent to which there are formed or remain complexes between the moiety and at least one of the substrate analogue and the substrate;
Specification