EP0194472A2
Expired / Lapsed
EP
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Fluorescence polarization assay for macromolecular hydrolases.
Assignee: Abbott Laboratories
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Filing: 1986-02-18
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Granted: —
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Expires: 2006-02-18
Abstract
Abstract A fluorescence polarization assay for determining the presence and/or amount of macromolecular hydrolases such as polysaccharide hydrolases, amylase, trypsin, proteases and lipases is disclosed. The disclosure provides reagents, processes for making the reagents useful in the assay and processes for conducting the assay. The assay involves the use of a substrate capable of being cleaved by the macromolecular hydrolase being determined, wherein the substrate is coupled to a fluorophore.
Inventors
- 👤 Mark Raymond Shaffar
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-26 23:34
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/37; C12Q1/40; C12Q1/44
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
35 words
A compound for use in a fluorescence polarization assay for the determination of macromolecular hydrolases, comprising: a substrate capable of being cleaved by a macromolecular hydrolase being determined; and a fluorophore coupled to said substrate.
Claim 2
Dependent Claim
35 words
A compound for use in a fluorescence polarization assay for the determination of macromolecular hydrolases, comprising: a substrate capable of being cleaved by a macromolecular hydrolase being determined; and a fluorophore coupled to said substrate.
Claim 3
Dependent Claim
23 words
The compound of Claim 1 wherein the substrate comprises amylose, starch, gelatin, hemoglobin, casein or lipoprotein, and the fluorophore comprises a fluorescein moiety.
Claim 4
Dependent Claim
26 words
The compound of Claim 2 wherein the fluorescein moiety comprises 4-chlorotriazin-2-(yl) amino fluorescein, 4,6-dichlorotriazin-2-(yl) amino fluorescein or derivatives thereof, thiocarbamyl fluorescein, fluorescein isothiocyanate or derivatives thereof.
Claim 5
Dependent Claim
82 words
A process for determining the activity of-a macromolecular hydrolase, comprising the steps of: a) preparing a test solution comprising a sample suspected of containing the macromolecular hydrolase, and a fluorophore-labelled substrate capable of being cleaved by the macromolecular hydrolase; b) incubating the test solution for a period of time; c) passing plane polarized light through the test solution of step b); and d) detecting the fluorescence polarization response from step c) as an indication of the macromolecular hydrolase activity in the sample.
Claim 6
Dependent Claim
82 words
A process for determining the activity of-a macromolecular hydrolase, comprising the steps of: a) preparing a test solution comprising a sample suspected of containing the macromolecular hydrolase, and a fluorophore-labelled substrate capable of being cleaved by the macromolecular hydrolase; b) incubating the test solution for a period of time; c) passing plane polarized light through the test solution of step b); and d) detecting the fluorescence polarization response from step c) as an indication of the macromolecular hydrolase activity in the sample.
Claim 7
Dependent Claim
22 words
The process of Claim 4 wherein the substrate comprises amylose, gelatin, hemoglobin, casein or lipoprotein and is coupled to a fluorescein moiety.
Claim 8
Dependent Claim
26 words
The process of Claim 5 wherein the substrate is coupled to 4,6-dichlorotriazin-2-(yl) aminofluorescein, a derivative of 4,6-dichlorotriazin-2-(yl) aminofluorescein, fluorescein isothiocyanate or a derivative of fluorescein isothiocyanate.
Claim 9
Dependent Claim
11 words
The process of Claim 4 wherein the macromolecular hydrolase is amylase.
Claim 10
Dependent Claim
11 words
The process of Claim 4 wherein the macromolecular hydrolase is trypsin.
Claim 11
Dependent Claim
11 words
The process of Claim 4 wherein the macromolecular hydrolase is lipase.
Claim 12
Dependent Claim
42 words
A process for making a reagent for a fluorescence polarization assay for determining a macromolecular hydrolase, comprising: coupling a substrate capable of being cleaved by the macromolecular hydrolase with a fluorophore capable of producing a fluorescence polarization response to plane polarized light.
Claim 13
Dependent Claim
42 words
A process for making a reagent for a fluorescence polarization assay for determining a macromolecular hydrolase, comprising: coupling a substrate capable of being cleaved by the macromolecular hydrolase with a fluorophore capable of producing a fluorescence polarization response to plane polarized light.
References Cited (Backward Citations)
No citation records indexed.
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