Patent US5356780A successfully refreshed from Google Patents.
US5356780A
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Determination of substrate of coenzyme requiring enzyme
Assignee: Merck Serono SA
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Filing: 1991-10-14
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Granted: 1994-10-18
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Expires: 2011-10-14
Abstract
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.
Inventors
- 👤 Grenville A. Robinson
- 👤 John G. Hurrell
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 12:06
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N33/53; G01N33/536; G01N33/542; G01N33/543; G01N33/54366; G01N33/54373
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
133 words
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.
Claim 2
Independent Claim
133 words
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.
Claim 3
Dependent Claim
21 words
A method as claimed in claim 1 wherein component (ii) comprises an apoenzyme which is not bound to its functional coenzyme.
Claim 4
Dependent Claim
33 words
A method as claimed in claim 2 wherein the apoenzyme is the apoenzyme of an oxidoreductase enzyme selected from the group consisting of alcohol dehydrogenase, cholesterol oxidase, lactic acid dehydrogenase and pyruvate dehydrogenase.
Claim 5
Dependent Claim
16 words
A method as claimed in claim 2 wherein the apoenzyme is the apoenzyme of glucose oxidase.
Claim 6
Dependent Claim
15 words
A method as claimed in claim 1 wherein component (ii) comprises a partly denatured enzyme.
Claim 7
Dependent Claim
16 words
A method as claimed in claim 1 wherein component (ii) comprises a genetically engineered deactivated enzyme.
Claim 8
Dependent Claim
32 words
A method as claimed in claim 1 wherein one of components (i) and (ii) is immobilised on a solid support, and the other of components (i) and (ii) carries a detectable label.
Claim 9
Dependent Claim
22 words
A method as claimed in claim 7 wherein the solid support is an optical waveguide and the detectable label is a fluorophore.
Claim 10
Dependent Claim
27 words
A method as claimed in claim 7, wherein the support is a part of a fluorescence capillary fill device and said determining is effected by detecting fluorescence.
Claim 11
Dependent Claim
18 words
A method as claimed in claim 1, wherein said determining comprises an optical technique employing surface plasmon resonance.
References Cited (Backward Citations)
US4978503A
Cited
US4810635A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-10-17 | 1998-04-18 | 1998-10-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-10-17 | 2002-04-18 | 2002-10-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-10-17 | 2006-04-18 | 2006-10-18 | $7700.00 | Window Closed / Lapsed |