Patent US2525447A successfully refreshed from Google Patents.
Patent EP0144744A2 successfully refreshed from Google Patents.
EP0144744A2
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EP
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Specific binding assay based on enzyme cascade amplification
Assignee: Bayer Corp
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Filing: 1984-11-03
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Granted: —
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Expires: 2004-11-03
Abstract
Abstract The present invention provides specific binding assay methods and reagent systems for the quantitative or qualitative determination of an analyte in a liquid medium based on heterogeneous or homogeneous protocols. In particular, the invention relates to the use of specific binding assay techniques wherein the label is a participant in or a modulator of an enzymatic reaction whereby a first enzyme acts on a first substrate to produce a second enzyme. The second enzyme can be used to catalyze a detectable response or can act on a second substrate to produce a third enzyme. The label is chosen from the first substrate or a cofactor or modulator of the first enzyme. In one embodiment, the first substrate is a zymogen which when acted upon by the first enzyme produces a second enzyme. When the final enzyme product is used to catalyze the production of a detectable response proportional of the concentration of the analyte, amplification results providing a very sensitive assay.
Inventors
- 👤 Diane A. Blake
- 👤 Robert C. Boguslaski
- 👤 Robert J. Carrico
- 👤 Mark T. Skarstedt
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-25 11:36
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/531; G01N33/532; G01N33/535; G01N33/58; G01N33/581
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
126 words
A specific binding assay method for determining an analyte in a test sample, which method comprises the steps of: (a) combining the test sample with a reagent system including a detectant reagent which is incorporated with a label, resulting in the formation of a bound-form of the detectant reagent and an unbound-form of the reagent, and (b) determining the detectant reagent in the bound-form or in the unbound-form by means of the label as a function of the analyte in the test sample, characterized in that the label is a participant in an enzymatic reaction wherein a first enzyme acts on a first substrate to produce a second enzyme, the label being selected from the first substrate or a cofactor or modulator of the first enzyme.
Claim 2
Dependent Claim
126 words
A specific binding assay method for determining an analyte in a test sample, which method comprises the steps of: (a) combining the test sample with a reagent system including a detectant reagent which is incorporated with a label, resulting in the formation of a bound-form of the detectant reagent and an unbound-form of the reagent, and (b) determining the detectant reagent in the bound-form or in the unbound-form by means of the label as a function of the analyte in the test sample, characterized in that the label is a participant in an enzymatic reaction wherein a first enzyme acts on a first substrate to produce a second enzyme, the label being selected from the first substrate or a cofactor or modulator of the first enzyme.
Claim 3
Dependent Claim
21 words
The method of claim 1 characterized in that the second enzyme acts on a second substrate to produce a third enzyme.
Claim 4
Dependent Claim
56 words
The method of claim 1 or 2 characterized in that either the first substrate is a zymogen or the second enzyme is formed by the addition of a chemical group to the first substrate or by the cleavage of a chemical group from the first substrate, which addition or cleavage is catalyzed by the first enzyme.
Claim 5
Independent Claim
28 words
The method of any of claims 1 to 3 of the homogeneous type wherein the detectant reagent is measured without separation of the bound- and unbound- forms thereof.
Claim 6
Dependent Claim
135 words
The method of claim 4, which method-comprises the steps of: (a) forming a reaction mixture by combining the liquid medium with a reagent system including (i) a binding partner for the analyte, and (ii) the detectant reagent which comprises the analyte, or a binding analog of the analyte, incorporated with a label, whereby there results in the reaction mixture a bound-form of the detectant reagent, wherein the detectant reagent is bound to the binding partner, and an unbound-form of the detectant reagent, wherein the detectant reagent is not so bound, the label having a detectable characteristic which is measurably different when the detectant reagent is in the bound-form compared to the unbound-form, and (b) measuring the detectable characteristic of the label in the reaction mixture as a function of the analyte in the liquid medium.
Claim 7
Independent Claim
31 words
The method of any of claims 1 to 3 of the heterogeneous type wherein the bound- and unbound- forms of the detectant are separated and the label measured in one thereof.
Claim 8
Dependent Claim
129 words
A reagent system for use in the specific binding assay determination of an analyte in a test sample, which system comprises a detectant reagent which is incorporated with a label, the combination of the reagent system and the test sample resulting in the formation of a bound-form of the detectant reagent and an unbound-form of the detectant reagent, determination of the detectant reagent in the bound-form or in the unbound-form by means of the label being a function of the analyte in the test sample, characterized in that the label is a participant in an enzymatic reaction wherein a first enzyme acts on a first substrate to produce a second enzyme, the label being selected from the first substrate or a cofactor or a modulator of the first enzyme.
Claim 9
Dependent Claim
129 words
A reagent system for use in the specific binding assay determination of an analyte in a test sample, which system comprises a detectant reagent which is incorporated with a label, the combination of the reagent system and the test sample resulting in the formation of a bound-form of the detectant reagent and an unbound-form of the detectant reagent, determination of the detectant reagent in the bound-form or in the unbound-form by means of the label being a function of the analyte in the test sample, characterized in that the label is a participant in an enzymatic reaction wherein a first enzyme acts on a first substrate to produce a second enzyme, the label being selected from the first substrate or a cofactor or a modulator of the first enzyme.
Claim 10
Dependent Claim
22 words
The reagent system of claim 7 characterized in that the second enzyme acts on a second substrate to produce a third enzyme.
Claim 11
Dependent Claim
56 words
The reagent system of claims 7 or 8 characterized in that either the substrate is a zymogen or the second enzyme is formed by the addition of a chemical group to the first substrate or by the cleavage of a chemical group from the first substrate, which addition or cleavage is catalyzed by the first enzyme.
Claim 12
Dependent Claim
59 words
The reagent system of any of claims 7 to 9 in the form of a test kit for use in the homogeneous immunoassay determination of an analyte in a liquid medium, comprising: (a) a binding partner for the analyte, and (b) a detectant reagent comprising the analyte, or a binding analog of the analyte, incorporated with the label. MS-131]
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