EP0158746A2
Expired / Lapsed
EP
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Visualization method for the direct or indirect detection of the reaction between a specific binding agent and the corresponding acceptor substance in blot overlay assays
Assignee: Janssen Pharmaceutica NV
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Filing: 1984-10-31
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Granted: —
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Expires: 2004-10-31
Abstract
Abstract A method for detecting and/or determining an agglomerate formed by the reaction between a specific binding agent and the corresponding acceptor substance according to the general methodology of blot overlay assays by using colloidal metal particles labelled components which may be visualized as a coloured signal at the surface of the blotting medium characteristic for the colloidal metal particles used or quantitatively determined following art-known spectopho- tometric procedures such as densitometry.
Inventors
- 👤 Marc K. J. J. Moeremans
- 👤 Guido F. T. Daneels
- 👤 Jan R. De Mey
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 12:40
CPC Classifications
C; C12; C12Q; C12Q1/00; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/558; G01N33/559; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/807; Y10S436/824
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
100 words
A process for detecting and/or determinating an agglomerate formed by the reaction between a specific binding agent and the corresponding acceptor substance according to the general methodology of blot overlay assays by using labelled components characterized in that said labelled components, obtained by coupling the desired component of said reaction to colloidal metal particles or by coupling a specific binding agent for the said component of said reaction to colloidal metal particles, are visualized as a coloured signal localized at the reaction site at the surface of the blotting medium or quantitatively determined at this site following art-known spectrophotometric procedures.
Claim 2
Dependent Claim
100 words
A process for detecting and/or determinating an agglomerate formed by the reaction between a specific binding agent and the corresponding acceptor substance according to the general methodology of blot overlay assays by using labelled components characterized in that said labelled components, obtained by coupling the desired component of said reaction to colloidal metal particles or by coupling a specific binding agent for the said component of said reaction to colloidal metal particles, are visualized as a coloured signal localized at the reaction site at the surface of the blotting medium or quantitatively determined at this site following art-known spectrophotometric procedures.
Claim 3
Dependent Claim
16 words
A process according to claim 1 wherein the specific binding agent is a specific binding protein.
Claim 4
Dependent Claim
51 words
A process according to claims 1 and 2, comprising the steps of i) immobilizing the acceptor substance to an immobilizing matrix; ii) contacting said immobilizing matrix with colloidal metal particles labelled specific binding agent; and iii) detecting the coloured signal at the reaction site at the surface of the immobilizing matrix.
Claim 5
Dependent Claim
62 words
A process accoring to claims 1 and 2, comprising the steps of i) immobilizing the acceptor substance to an immobilizing matrix; ii) contacting said immobilizing matrix with an unlabelled specific binding agent and, subsequently, colloidal metal labelled protein specific for the unlabelled specific binding protein; and iii) detecting the coloured signal at the reaction site at the surface of the immobilizing matrix.
Claim 6
Dependent Claim
55 words
A process according to claims 3 and 4 wherein the immobilization- step is effected by direct adsorption and/or covalent binding, optionally after applying a procedure of electrophoretic separation and applying a procedure of transfer or blotting from the electrophoretic medium to the blotting medium and subsequently quenching remaining agent specific binding sites by art-known procedures.
Claim 7
Dependent Claim
48 words
A process according to claim 3 and 4 wherein the immobilization- step is effected by allowing the acceptor substance to become bound by a specific binding protein which has become immobilized to the blotting medium, by contacting said blotting medium with an aqueous solution.which contains the acceptor substance.
Claim 8
Dependent Claim
37 words
A process according to claims 1-6 wherein the colloidal metal particles consist of gold, silver, platinum or compounds of these metals or iron or copper compounds having a particle size comprised between 3 nm and 100 nm.
Claim 9
Dependent Claim
27 words
A process according to claims 1-7 wherein the colloidal metal particles consist of gold and/or silver having a particle size comprised between 3 nm and 100 nm.
Claim 10
Dependent Claim
22 words
A process according to claims 1-8 wherein the final detection of the colloidal metal particles is effected after applying a physical developer.
Claim 11
Dependent Claim
14 words
A process according to claim 9 wherein the physical developer is a silver-containing compound.
Claim 12
Dependent Claim
79 words
A test kit to be used for the direct or indirect determination of one of the components of the reaction between a specific binding agent and the corresponding acceptor substance according to general methods of blot overlay assay as described in any of claims 1 to 10 containing a colloidal metal particle labelled component obtained by coupling a component of the said reaction or a component that can be used to detect this reaction indirectly to colloidal metal particles.
References Cited (Backward Citations)
No citation records indexed.
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