Patent EP0421788A2 successfully refreshed from Google Patents.
EP0421788A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Haloperoxidase-acid-optimum chemiluminescence assay system
Assignee: Exoxemis Inc
•
Filing: 1990-10-04
•
Granted: —
•
Expires: 2010-10-04
Abstract
Abstract A highly sensitive chemiluminescent indicator system is disclosed for determining the presence or amount of an analyte in a liquid sample. The new acid optimum chemiluminescent indicator system comprises haloperoxidase (halide:hydrogen peroxide oxidoreductase), halide, oxidant and chemiluminigenic substrate. The indicator system acts as a synthesizer of highly reactive singlet molecular oxygen (¹O₂), which reacts with the chemiluminigenic substrate to yield an excited state, oxidized reaction product. The excited state reaction product then relaxes to a lower energy (e.g., ground) state with the emission of measurable light in an amount related to the amount of each of the reaction participants present in a reaction solution. Known, non-rate limiting amounts of three of the reaction participants are provided in an assay solution to determine the presence or amount of the fourth participant in a test sample. The fourth participant in the test sample may be the analyte of interest, or may be produced or consumed in the test sample through one or more preliminary reactions involving the ultimate analyte of interest, with the amount of the fourth participant being related to the amount of analyte in the test solution. Accordingly, the indicator system of the invention may be employed for the determination of a wide variety of analytes. The indicator system operates most efficiently over the range from acid to slightly basic pH, e.g., at a pH of about 3 to about 8. Also described is a variety of illustrative assay formats in which the indicator reaction of the invention may be employed, as well as kits for use in carrying out assays utilizing the haloperoxidase/halide/oxidant/substrate indicator system.
Inventors
- 👤 Robert Charles Allen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 13:17
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
144 words
A method for determining the presence or amount of an analyte selected from the group comprising: peroxide, halide, haloperoxidase enzyme, chemiluminigenic substrate for said enzyme or an analyte capable of producing or "consuming peroxide, halide, haloperoxidase or chemiluminigenic substrate in one or more preliminary reactions, in a sample suspected of containing an unknown amount of the analyte, the method comprising a) contacting the sample with an assay solution comprising a known, non-rate limiting amount of the other members of the said group; b) maintaining the pH of the assay solution less than about 8; c) measuring at least one characteristic of light emitted by the assay solution; and d) comparing the said characteristic of the light measured with that of a standard solution containing a known amount of the analyte as a measure of the presence or amount of the analyte in the sample.
Claim 2
Independent Claim
144 words
A method for determining the presence or amount of an analyte selected from the group comprising: peroxide, halide, haloperoxidase enzyme, chemiluminigenic substrate for said enzyme or an analyte capable of producing or "consuming peroxide, halide, haloperoxidase or chemiluminigenic substrate in one or more preliminary reactions, in a sample suspected of containing an unknown amount of the analyte, the method comprising a) contacting the sample with an assay solution comprising a known, non-rate limiting amount of the other members of the said group; b) maintaining the pH of the assay solution less than about 8; c) measuring at least one characteristic of light emitted by the assay solution; and d) comparing the said characteristic of the light measured with that of a standard solution containing a known amount of the analyte as a measure of the presence or amount of the analyte in the sample.
Claim 3
Dependent Claim
16 words
A method as claimed in claim 1 wherein the assay solution is a homogeneous assay solution.
Claim 4
Dependent Claim
77 words
A method as claimed in claim 1 wherein the assay solution is a heterogeneous assay solution and the reagent system further comprises an insoluble phase, a first specific binding substance immobilized on the insoluble phase and having binding specificity for the analyte, and a second specific binding substance conjugated either to the haloperoxidase, or to an oxidase capable of generating peroxide, or to the chemiluminigenic substrate, said second specific binding substance having binding specificity for the analyte.
Claim 5
Dependent Claim
49 words
A method as claimed in claim 3 which further comprises contacting the immobilized first specific binding substance with the sample and with the second specific binding substance, separating the soluble phase from the insoluble phase, and measuring the characteristic of the light in the soluble or the insoluble phase.
Claim 6
Independent Claim
41 words
A method as claimed in any of claims 1-4 which further comprises adjusting the pH of the assay solution to a pH in the range of about 3 to about 8 prior to measuring the said characteristic of the emitted light.
Claim 7
Independent Claim
30 words
A method as claimed in any one of claims 1-5 wherein the analyte is peroxide or an analyte capable of producing or consuming peroxide in one or more preliminary reactions.
Claim 8
Dependent Claim
35 words
A method as claimed in claim 6 wherein the peroxide is produced in the sample in one or more preliminary reactions in which the peroxide is a reaction product and the analyte is a reactant.
Claim 9
Dependent Claim
30 words
A method as claimed in claim 6 wherein the peroxide is consumed in the sample in one or more preliminary reactions in which the peroxide and the analyte are reactants.
Claim 10
Independent Claim
30 words
A method as claimed in any of claims 1-5 wherein the analyte is a halide, or an analyte capable of producing or consuming halide in one or more preliminary reactions.
Claim 11
Independent Claim
14 words
A method as claimed in any of claims 1-5 wherein the analyte is haloperoxidase.
Claim 12
Dependent Claim
51 words
A kit for use in a chemiluminescent or chemiluminometric assay comprising at least three of the following: a) a haloperoxidase; b) a peroxide; c) a halide; d) a chemiluminigenic substrate; and e) a buffer solution for maintaining the pH of the assay within the range of about 3 to about 8.
Claim 13
Dependent Claim
51 words
A kit for use in a chemiluminescent or chemiluminometric assay comprising at least three of the following: a) a haloperoxidase; b) a peroxide; c) a halide; d) a chemiluminigenic substrate; and e) a buffer solution for maintaining the pH of the assay within the range of about 3 to about 8.
Claim 14
Dependent Claim
21 words
A method as claimed in claims 1-10 or a kit as claimed in claim 11 wherein the peroxide is hydrogen peroxide.
Claim 15
Independent Claim
25 words
A method or a kit as claimed in any preceding claim wherein the haloperoxidase is myeloperoxidase (MPO), eosinophil peroxidase (EPO), lactoperoxidase (LPO) or chloroperoxidase (CPO).
Claim 16
Dependent Claim
24 words
A method or a kit as claimed in claim 13 wherein the haloperoxidase is MPO or CPO and the halide is bromide or chloride.
Claim 17
Dependent Claim
42 words
A method or a kit as claimed in claim 14 wherein the sample is a biological fluid and the amount of MPO in the sample is determined as a measure of the amount of polymorphonuclear leukocytes or blood monocytes in the sample.
Claim 18
Dependent Claim
22 words
A method or a kit as claimed in Claim 13 wherein the haloperoxidase is EPO or LPO and the halide is bromide.
Claim 19
Dependent Claim
38 words
A method or a kit as claimed in Claim 16 wherein the sample is a biological fluid and the amount of EPO in the sample is determined as a measure of the amount of eosinophils in the sample.
Claim 20
Independent Claim
18 words
A method or a kit as claimed in any preceding claim wherein the halide is bromide or chloride.
Claim 21
Dependent Claim
57 words
A method as claimed in claim 9 wherein the analyte is chloride, and which further comprises determining the MPO- or CPO-dependent characteristics of the light emitted by the assay system minus the EPO- or LPO-dependent characteristics of the light emitted by the assay system as a measure of the presence or amount of chloride in the sample.
Claim 22
Dependent Claim
58 words
A method as claimed in claim 9 wherein the analyte is bromide, and which further comprises determining the EPO- or LPO-dependent characteristics of the light emitted by the assay system divided by the MPO- or CPO-dependent characteristics of the light emitted by the assay system as a measure of the presence or amount of bromide in the sample.
Claim 23
Independent Claim
36 words
A method or a kit as claimed in any preceding claims wherein the chemiluminigenic substrate is a cyclic hydrazide or a dioxetane precursor capable of reacting with singlet-multiplicity oxygen to produce a dioxetane or a dioxetanone.
Claim 24
Dependent Claim
19 words
A method as claimed in claim 21 wherein the cyclic hydrazide is a 2,3-dihydro-1,4-phthalazinedione such as luminol or isoluminol.
Claim 25
Dependent Claim
47 words
The use of three of the following agents: a) a haloperoxidase; b) a peroxide; c) a halide; or d) a chemiluminescent substrate or an agent capable of producing any of these in the preparation of an assay solution for the detection of the fourth of the agents.
Claim 26
Dependent Claim
47 words
The use of three of the following agents: a) a haloperoxidase; b) a peroxide; c) a halide; or d) a chemiluminescent substrate or an agent capable of producing any of these in the preparation of an assay solution for the detection of the fourth of the agents.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.