Patent EP0247796A1 successfully refreshed from Google Patents.
EP0247796A1
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Solid phase immunoassay method
Assignee: Unilever PLC
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Filing: 1987-05-21
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Granted: —
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Expires: 2007-05-21
Abstract
Abstract An immunoassay for determining qualitatively or quantitatively the presence of an analyte in a sample by means of specific binding, wherein the sample is contacted with a moveable solid phase carrier material, e.g. magnetic particles (620) on which is immobilised a first binding reagent having specificity for the analyte and with a labelled reagent which can participate in either a "sandwich" or a "competition" reaction with the first reagent in the presence of the analyte and following an incubation period sufficient to allow the reaction to take place, the carrier material is moved within the assay medium (618) to a location adjacent a signal sensing means (602), the label generating a signal, such as chemiluminescent light, continuously in the assay medium and the magnitude of the signal generated in the vicinity of the sensing means being used as a measure of the extent to which the binding reaction has occurred. Preferably the assay medium incorporates a masking or quenching agent which suppresses signal generated in regions of the assay medium remote from the signal sensing means.
Inventors
- 👤 Paul James Davis
- 👤 Rosemary Ann Lucy Drake
- 👤 William Edward Hornby
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 06:39
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N33/53; G01N33/543; G01N33/54313; G01N33/54326; G01N35/00; G01N2035/00465; G01N2035/00534; G01N35/10; G01N35/1009; G01N2035/1025; G01N35/0098
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
154 words
An assay method for determining qualitatively and/or quantitatively the presence of an analyte in a sample by means of specific binding, wherein the sample is contacted with a moveable solid phase carrier material, such as a particulate carrier, on which is immobilised a first binding reagent having specificity for the analyte and with a labelled specific binding reagent which can participate in either a "sandwich" or a "competition" reaction with the first reagent in the presence of the analyte, and in which method, following an incubation period sufficient to allow the reaction to take place, the moveable solid phase carrier material is moved within the assay medium to a location adjacent a signal sensing means, the label generating a signal continuously in the assay medium and the magnitude of the signal generated in the vicinity of the sensing means being used as a measure of the extent to which the binding reaction has occurred.
Claim 2
Dependent Claim
154 words
An assay method for determining qualitatively and/or quantitatively the presence of an analyte in a sample by means of specific binding, wherein the sample is contacted with a moveable solid phase carrier material, such as a particulate carrier, on which is immobilised a first binding reagent having specificity for the analyte and with a labelled specific binding reagent which can participate in either a "sandwich" or a "competition" reaction with the first reagent in the presence of the analyte, and in which method, following an incubation period sufficient to allow the reaction to take place, the moveable solid phase carrier material is moved within the assay medium to a location adjacent a signal sensing means, the label generating a signal continuously in the assay medium and the magnitude of the signal generated in the vicinity of the sensing means being used as a measure of the extent to which the binding reaction has occurred.
Claim 3
Dependent Claim
107 words
An assay method according to claim 1, which utilises a specific binding reagent which is labelled such that the label continuously generates a signal in the assay medium, which signal is suppressed while the labelled binding reagent is uniformly dispersed in the assay medium, and in which method, following the binding reaction, the particulate carrier material is localised in an environment having insufficient signal-suppressing, ability to overcome the locally-generated signal, and hence a detectable signal can be generated the nature and/or magnitude of which is dependent on the extent of binding that has taken place between the analyte and the specific binding reagent immobilised on the particles.
Claim 4
Dependent Claim
26 words
An assay method according to claim 1 or claim 2, wherein the signal detected by the sensing means is visible light originating from a chemiluminescent reaction.
Claim 5
Dependent Claim
29 words
An assay method according to claim 3, wherein the label generates hydrogen peroxide in the assay medium and the peroxide is detected via its participation in a chemiluminescent reaction.
Claim 6
Dependent Claim
20 words
An assay method according to claim 4, wherein the label is a peroxide-destroying enzyme which engages in a chemiluminescent reaction.
Claim 7
Dependent Claim
26 words
An assay method according to claim 1 or claim 2, wherein the label generates hydrogen peroxide in the assay medium and the peroxide is measured electrochemically.
Claim 8
Dependent Claim
37 words
An assay method according to any one of the preceding claims, wherein the assay medium incorporates a masking or quenching agent which suppresses signal generated in regions of the assay medium remote from the signal sensing means.
Claim 9
Dependent Claim
46 words
An assay method according to claim 7, wherein the detectable signal is chemiluminescent light and the masking or quenching reagent renders the assay medium opaque or at least insufficiently translucent for light from regions remote from the signal sensing means to reach the signal sensing means.
Claim 10
Dependent Claim
27 words
An assay method according to claim 7, wherein the masking or quenching reagent is a chemical reagent which consumes an essential component of the signal generating system.
Claim 11
Dependent Claim
24 words
An assay method according to claim 9, wherein the label produces hydrogen peroxide in the assay medium and the consumer reagent is catalase enzyme.
Claim 12
Dependent Claim
50 words
An assay method according to claim 5, wherein the label is horseradish peroxidase and the assay medium incorporates a peroxide, such as hydrogen peroxide, and a chemiluminescent 2,3-dihydro-1,4-phthalazinedione, such as luminol, and the assay medium optionally also incorporates an enhancer for the chemiluminescent reaction, such as para-iodophenol, para-phenylphenol and/or 2-chloro-4-phenylphenol.
Claim 13
Dependent Claim
156 words
An assay method according to claim 1, wherein the assay medium incorporates: a) a first population of localisable particles on which is immobilised a binding reagent having specificity for the analyte; b) a second specific binding reagent labelled such that it continuously produces within the assay medium a precursor for a chemiluminescent reaction; c) a consumer reagent dispersed in the test medium which competes effectively for the precursor and thereby inhibits the production of chemiluminescence while the labelled specific binding reagent remains dispersed in the assay medium; and d) a second population of localisable particles bearing or containing a reagent which can react with the precursor to generate chemiluminescent light; in which assay method co-localisation of the two particle populations adjacent a light detection means enables observation of chemiluminescence produced in an amount dependant on the extent to which the specific binding reagent immobilised on the first population of particles has become bound to the analyte.
Claim 14
Dependent Claim
56 words
An assay method according to claim 12, wherein the label is glucose oxidase, the assay medium contains glucose and a chemiluminescent 2,3-dihydro-1,4-phthalazinedione such as luminol, the consumer reagent is catalase enzyme, the reagent associated with the second population of localisable particles is horseradish peroxidase, and the assay medium optionally also an enhancer such as incorporates para-iodophenol.
Claim 15
Dependent Claim
32 words
An assay method according to claim 1, wherein a chemical signal is detected by localising a particulate carrier adjacent a surface containing or carrying a reagent which interacts with the chemical signal.
Claim 16
Dependent Claim
37 words
An assay method according to claim 14, wherein the chemical signal is hydrogen peroxide and the surface is a gel body containing horseradish peroxidase and a chemiluminescent 2,3-dihydro-1,4-phthalazinedione luminol, and optionally also an enhancer such as para-iodophenol.
Claim 17
Dependent Claim
19 words
An assay method according to any one of the preceding claims, applied to the screening of cell culture media.
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