US4830959A
Expired / Lapsed
US
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Electrochemical enzymic assay procedures
Assignee: Medisense Inc
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Filing: 1986-11-10
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Granted: 1989-05-16
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Expires: 2006-11-10
Abstract
Abstract Electrochemical detection of mediator level is employed in a method of assay using a redox enzyme and redox substrate to detect conversion to an effective mediator by an assay enzyme label of a compound which is non-mediating under the assay conditions.
Inventors
- 👤 Calum J. McNeil
- 👤 Joseph V. Bannister
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 07:56
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/581; G01N33/53; G01N33/543; G01N33/54366; G01N33/54373; G01N33/5438; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/817; Y10S436/00; Y10S436/806; Y10S436/904
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
183 words
A method of assaying for activity of a first enzyme able to convert a non-mediator to a mediator, said mediator having a reduced form and an oxidized form, said method comprising: (a) providing a sample to be assayed for said activity of said first enzyme; (b) contacting said sample with freely circulating non-mediator; (c) providing a second enzyme, said second enzyme being catalytically active in the absence of said mediator; (d) providing a substrate for said second enzyme; (e) providing an electrode at an electrical potential which is intermediate between the redox potential of said mediator and said non-mediator, said mediator being capable of transferring charge between said second enzyme and said electrode, said electrical potential is such that said mediator is converted from said reduced form to said oxidized form at said electrode and said non-mediator is not oxidized or reduced at said electrode; (f) measuring the current generated at said electrode, said current providing a measure of the conversion of said non-mediator to said mediator; and (g) correlating the current generated at said electrode with the presence of said first enzyme.
Claim 2
Independent Claim
183 words
A method of assaying for activity of a first enzyme able to convert a non-mediator to a mediator, said mediator having a reduced form and an oxidized form, said method comprising: (a) providing a sample to be assayed for said activity of said first enzyme; (b) contacting said sample with freely circulating non-mediator; (c) providing a second enzyme, said second enzyme being catalytically active in the absence of said mediator; (d) providing a substrate for said second enzyme; (e) providing an electrode at an electrical potential which is intermediate between the redox potential of said mediator and said non-mediator, said mediator being capable of transferring charge between said second enzyme and said electrode, said electrical potential is such that said mediator is converted from said reduced form to said oxidized form at said electrode and said non-mediator is not oxidized or reduced at said electrode; (f) measuring the current generated at said electrode, said current providing a measure of the conversion of said non-mediator to said mediator; and (g) correlating the current generated at said electrode with the presence of said first enzyme.
Claim 3
Dependent Claim
18 words
The method of claim 1 wherein said first enzyme is a label for an antibody or an antigen.
Claim 4
Dependent Claim
11 words
The method of claim 2, wherein said immunoassay is an ELISA.
Claim 5
Dependent Claim
42 words
The method of claim 1, 2, or 3 wherein said mediator is selected from the group consisting of: metallocenes, ruthenium compounds, carboranes, conductive salts of tetracyanoquinodimethane (TCNQ), haloanils, viologens, quinones, alkyl-substituted phenazines, and bis-cyclo pentadienyl (Cp) 2 MXx complexes of transition metals.
Claim 6
Dependent Claim
15 words
The method of claim 1, 2, or 3 wherein said mediator is a phenol derivative.
Claim 7
Dependent Claim
19 words
The method of claim 4 wherein said phenol derivative is selected from the group consisting of: ferrocene-phenols; and indophenols.
Claim 8
Dependent Claim
11 words
The method of claim 6, wherein said mediator is [N-ferrocenoyl]-4-aminophenyl phosphate.
Claim 9
Dependent Claim
12 words
The method of claim 1 wherein said first enzyme is a phosphatase.
Claim 10
Dependent Claim
22 words
The method of claim 1 wherein said electrode is poised at an electrical potential higher than the redox potential of said mediator.
Claim 11
Independent Claim
132 words
A system for detecting activity of a first enzyme in a sample, said first enzyme converting a non-mediator to a mediator, said mediator having a reduced form and an oxidized form, said system comprising: (a) said non-mediator which is capable of conversion by said first enzyme to said mediator; (b) a second enzyme which converts said mediator from said oxidized form to said reduced form; (c) a substrate for said second enzyme; and (d) an electrode and means for poising said electrode at an electrical potential which is intermediate between the redox potential of said mediator and said non-mediator, wherein said electrical potential is such that said mediator is converted from said reduced form to said oxidized form at said electrode and said non-mediator is not oxidized or reduced at said electrode.
Claim 12
Independent Claim
132 words
A system for detecting activity of a first enzyme in a sample, said first enzyme converting a non-mediator to a mediator, said mediator having a reduced form and an oxidized form, said system comprising: (a) said non-mediator which is capable of conversion by said first enzyme to said mediator; (b) a second enzyme which converts said mediator from said oxidized form to said reduced form; (c) a substrate for said second enzyme; and (d) an electrode and means for poising said electrode at an electrical potential which is intermediate between the redox potential of said mediator and said non-mediator, wherein said electrical potential is such that said mediator is converted from said reduced form to said oxidized form at said electrode and said non-mediator is not oxidized or reduced at said electrode.
Claim 13
Dependent Claim
40 words
The system of claim 10, said mediator being chosen from the group consisting of: metallocenes, ruthenium compounds, carboranes, conductive salts of TCNQ, haloanils, viologen, a quinone, an alkyl-substituted phenazine derivative, a bis-cyclo pentadienyl (Cp) 2 MXx complexes of transition metals.
Claim 14
Dependent Claim
11 words
The system of claim 10, said mediator being a phenol derivative.
Claim 15
Dependent Claim
19 words
The system of claim 12, said phenol derivative is chosen from the group consisting of: ferrocene-phenols and indophenol compounds.
Claim 16
Dependent Claim
11 words
The system of claim 13, wherein said mediator is [N-ferrocenoyl]-4-aminophenyl phosphate.
Claim 17
Dependent Claim
11 words
The system of claim 10, said first enzyme being a phosphatase.
Claim 18
Dependent Claim
18 words
The system of claim 10, wherein said electrical potential is higher than the redox potential of said mediator.
Claim 19
Dependent Claim
20 words
The system of claim 10, wherein said first enzyme is bound to an antigen or an antibody as a label.
References Cited (Backward Citations)
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