Patent US4809695A successfully refreshed from Google Patents.
EP0465577B1
Expired / Lapsed
EP
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Catalyzed reporter deposition
Assignee: Revvity Health Sciences Inc
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Filing: 1990-03-28
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Granted: 1994-03-16
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Expires: 2010-03-28
Abstract
Abstract The present invention concerns a method to catalyze reporter deposition to improve detection or quantitation of an analyte in a sample by amplifying the detector signal which comprises immobilizing an analyte dependent enzyme activation system which catalyzes deposition of reporter by activating a conjugate consisting of a detectably labeled substrate specific for the enzyme system, said conjugate reacts with the analyte dependent enzyme activation system to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system. In another embodiment the invention concerns an assay for detecting or quantitating the presence or absence of an analyte in a sample using catalyzed reporter deposition to amplify the reporter signal. Also described are novel compounds which can be used as reagents to prepare HABA-type conjugates.
Inventors
- 👤 Mark Norman Bobrow
- 👤 Richard Calvin Ebersole
- 👤 Gerald Joseph Litt
- 👤 John Richard Moran
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 04:42
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535; G01N33/536; G01N33/542; G01N33/58; G01N33/581
Claims Analysis
Total Claims: 31
Claim 1
Independent Claim
91 words
A method for the detection or quantitation of an analyte in an assay which comprises using an analyte dependent enzyme activation system comprising at least one enzyme to react with a conjugate consisting of a detectably labeled substrate specific for the enzyme system to form an activated conjugate which deposits substantially wherever at least one receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system, wherein deposited detectable labels either directly or indirectly generate a signal which can be detected or quantitated.
Claim 2
Dependent Claim
32 words
A method according to claim 1 wherein at least one enzyme of the analyte dependent enzyme activation system is selected from the group consisting of oxidoreductases, hydrolases, lyases, transferases, isomerases, and ligases.
Claim 3
Dependent Claim
22 words
A method according to claim 2 wherein the enzyme is selected from the group consisting of peroxidases, oxidases, phosphatases, esterases and glycosidases.
Claim 4
Dependent Claim
29 words
A method according to claim 3 wherein the enzyme is selected from the group consisting of horseradish peroxidase, glucose oxidase, alkaline phosphatase and beta-galactosidase, and preferably is horseradish peroxidase.
Claim 5
Dependent Claim
20 words
A method according to claim 4 wherein the conjugate is selected from the group consisting of biotin-tyramine, p-hydroxyphenylpropionylbiocytin, or fluorescein-tyramine.
Claim 6
Dependent Claim
15 words
A method according to claim 1 wherein the conjugate is reacted with detectably labeled antibody.
Claim 7
Dependent Claim
21 words
A method according to claim 1 wherein the conjugate is reacted with a detectably labeled member of a specific binding pair.
Claim 8
Dependent Claim
15 words
A method according to claim 1 wherein the conjugate is reacted with detectably labeled streptavidin.
Claim 9
Dependent Claim
33 words
A method according to claim 1 wherein the detectable label is selected from the group consisting of enzymes, radioactive isotopes, fluorogenic, chemiluminescent, or electrochemical materials or a member of a specific binding pair.
Claim 10
Dependent Claim
12 words
A method according to claim 1 wherein the conjugate is 6-(phenoxy-(4'-azo-2''-carboxyethylphenyl))-hexanoyl-alkaline phosphatase.
Claim 11
Dependent Claim
94 words
An assay for detecting or quantitating the presence or absence of an analyte in a sample which comprises a) immobilizing the analyte; b) reacting the product of step (a) with an analyte-dependent enzyme activation system; c) reacting the product of step (b) with a conjugate consisting of a detectably labeled substrate to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system; and d) detecting or quantitating the presence or absence of the analyte in the sample.
Claim 12
Dependent Claim
31 words
An assay according to claim 11 wherein the analyte dependent enzyme activation system has at least one enzyme selected from the group consisting of oxidoreductases, hydrolases, lyases, transferases, isomerases, and ligases.
Claim 13
Dependent Claim
22 words
An assay according to claim 12 wherein the enzyme is selected from the group consisting of peroxidases, oxidases, phosphatases, esterases and glycosidases.
Claim 14
Dependent Claim
24 words
An assay according to claim 13 wherein the enzyme is selected from the group consisting of horseradish peroxidase, glucose oxidase, alkaline phosphatase, and beta-galactosidase.
Claim 15
Dependent Claim
33 words
An assay according to claim 11 wherein the detectable label is selected from the group consisting of enzymes, radioactive isotopes, fluorogenic, chemiluminescent, or electrochemical materials or a member of a specific binding pair.
Claim 16
Dependent Claim
27 words
An assay according to claim 14 wherein the enzyme is horseradish peroxidase and the conjugate is selected from the group consisting of biotin tyramine, p-hydroxyphenylpropionyl-biocytin or fluorescein-tyramine.
Claim 17
Dependent Claim
12 words
A method according to claim 11 wherein the conjugate is 6-(phenoxy-(4'-azo-2''-carboxyethylphenyl))-hexanoyl-alkaline phosphatase.
Claim 18
Dependent Claim
21 words
An assay according to claim 11 wherein the conjugate is reacted with a detectably labeled member of a specific binding pair.
Claim 19
Dependent Claim
15 words
An assay according to claim 11 wherein the conjugate is reacted with detectably labeled antibody.
Claim 20
Dependent Claim
15 words
An assay according to claim 11 wherein the conjugate is reacted with detectably labeled streptavidin.
Claim 21
Dependent Claim
12 words
An assay according to claim 11 wherein said assay is an immunoassay.
Claim 22
Dependent Claim
78 words
An assay for detecting or quantitating the presence or absence of an analyte in a sample which comprises a) reacting an analyte dependent enzyme activation system with a conjugate consisting of a detectably labeled substrate to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system; and b) detecting or quantitating the presence or absence of the analyte in the sample.
Claim 23
Dependent Claim
31 words
An assay according to claim 22 wherein the analyte dependent enzyme activation system has at least one enzyme selected from the group consisting of oxidoreductases, hydrolases, lyases, transferases, isomerases, and ligases.
Claim 24
Dependent Claim
22 words
An assay according to claim 23 wherein the enzyme is selected from the group consisting of peroxidases, oxidases, phosphatases, esterases and glycosidases.
Claim 25
Dependent Claim
24 words
An assay according to claim 24 wherein the enzyme is selected from the group consisting of horseradish peroxidase, glucose oxidase, alkaline phosphatase, and beta-galactosidase.
Claim 26
Dependent Claim
33 words
An assay according to claim 22 wherein the detectable label is selected from the group consisting of enzymes, radioactive isotopes, fluorogenic, chemiluminescent, or electrochemical materials or a member of a specific binding pair.
Claim 27
Dependent Claim
21 words
An assay according to claim 22 wherein the conjugate is reacted with a detectably labeled member of a specific binding pair.
Claim 28
Dependent Claim
15 words
An assay according to claim 22 wherein the conjugate is reacted with detectably labeled streptavidin.
Claim 29
Dependent Claim
15 words
An assay according to claim 22 wherein the conjugate is reacted with detectably labeled antibody.
Claim 30
Dependent Claim
12 words
An assay according to claim 22 wherein said assay is an immunoassay.
Claim 31
Dependent Claim
29 words
Process for preparing 6-(phenoxy-(4'-azo-2''-carboxyethylphenyl))-hexanoyl-alkaline phosphatase, which comprises 1) dissolving (N-succinimidyl)-6-(phenoxy-(4'-azo-2''-carboxy-ethylphenyl))hexanoate in a minium volume of dimethylsulfoxide, and 2) adding said solution to a buffered aqueous solution of alkaline phosphatase.
References Cited (Backward Citations)
No citation records indexed.
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