US4629696A
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Process for the selective production of reduced oxygen species
Assignee: Roche Diagnostics GmbH
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Filing: 1983-09-30
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Granted: 1986-12-16
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Expires: 2003-09-30
Abstract
Abstract Process for the selective production of the reduced oxygen species superoxide, hydrogen peroxide and hydroxyl radicals, wherein oxygen is reduced with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an appropriate autoxidisable redox partner and optionally of an appropriate buffer system. Also, process for the determination of superoxide dismutase in which a superoxide-yielding reaction is coupled with two competing superoxide-consuming reactions, one superoxide-consuming reaction being a conventional indicator reaction and the other superoxide-consuming reaction being the superoxide dismutase reaction, wherein, as superoxide-yielding reaction, there is used the reduction of oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an autoxidisable one-electron step-inducing redox partner with a one-electron redox potential in the range of from -150 mV to -500 mV and optionally of an appropriate buffer system. Also, process for the determination of NAD(P)H and of NAD(P)H-yielding reactions in which oxygen is reduced to hydrogen peroxide by NAD(P)H and the hydrogen peroxide is determined in knowm manner, wherein the reduction of the oxygen by NAD(P)H is carried out in the presence of an NAD(P)H-dependent, non-autoxidisable diaphorase, of an appropriate two-electron redox partner with a two-electron redox potential in the range of from 0 mV to 150 mV and of an appropriate buffer system. Also, reagents for carrying out these processes.
Inventors
- 👤 Erich Elstner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 11:23
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
35 words
A process for the selective production of the reduced oxygen species superoxide, hydrogen peroxide or hydroxyl radicals, comprising reducing oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase and an autoxidizable redox partner.
Claim 2
Dependent Claim
35 words
A process for the selective production of the reduced oxygen species superoxide, hydrogen peroxide or hydroxyl radicals, comprising reducing oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase and an autoxidizable redox partner.
Claim 3
Dependent Claim
17 words
The process of claim 1, wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is NADP-ferredoxin-(Cyt c)-oxidoreductase or diaphorase from microorganisms.
Claim 4
Dependent Claim
35 words
The process of claim 1 to produce superoxide, wherein the autoxidizable redox partner is a one-electron step-inducing substance with a one-electron redox potential E o in the range of from -150 mV to -500 mV.
Claim 5
Dependent Claim
33 words
The process of claim 2 to produce superoxide wherein the autoxidizable redox partner is a bipyridilium salt, a triazonium salt, an anthraquinone derivative, a nitrofuran derivative, a ferredoxin, a pteridine or an isoalloxazine.
Claim 6
Dependent Claim
38 words
The process of claim 1 for the production of hydrogen peroxide wherein the autoxidizable redox partner is a two-electron step-inducing substance with a two-electron redox potential E o in the range of from 0 mV to 150 mV.
Claim 7
Dependent Claim
18 words
The process of claim 2 to produce hydrogen peroxide, wherein the autoxidizable redox partner is a quinone derivative.
Claim 8
Dependent Claim
42 words
The process of claim 1 to produce hydroxyl radicals, wherein the autoxidizable redox partner is a one-electron step-inducing substance with a one-electron redox potential E o in the range of from -150 mV to -500 mV in the presence of superoxide dismutase.
Claim 9
Dependent Claim
49 words
A process for the selective production of hydroxyl radicals comprising reducing hydrogen peroxide with NAD(P)H, under anaerobic conditions in the presence of NAD(P)H-dependent, non-autoxidizable diaphorase and an autoxidizable, one-electron step-inducing redox partner with a one-electron redox potential E o in the range of from -150 mV to -500 mV.
Claim 10
Dependent Claim
49 words
A process for the selective production of hydroxyl radicals comprising reducing hydrogen peroxide with NAD(P)H, under anaerobic conditions in the presence of NAD(P)H-dependent, non-autoxidizable diaphorase and an autoxidizable, one-electron step-inducing redox partner with a one-electron redox potential E o in the range of from -150 mV to -500 mV.
Claim 11
Dependent Claim
16 words
The process of claim 1 wherein the reaction is buffered to the range of pH 6-9.
Claim 12
Dependent Claim
36 words
A reagent for the production of the reduced oxygen species superoxide, hydrogen peroxide or hydroxyl radicals by the reduction of oxygen or hydrogen peroxide with NAD(P)H, comprising NAD(P)H, an NAD(P)H-dependent, non-autoxidizable diaphorase and autoxidizable redox partner.
Claim 13
Dependent Claim
36 words
A reagent for the production of the reduced oxygen species superoxide, hydrogen peroxide or hydroxyl radicals by the reduction of oxygen or hydrogen peroxide with NAD(P)H, comprising NAD(P)H, an NAD(P)H-dependent, non-autoxidizable diaphorase and autoxidizable redox partner.
Claim 14
Dependent Claim
17 words
The reagent of claim 10 wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is NADP-ferredoxin-Cyt c)-oxidoreductase or diaphorase from microorganisms.
Claim 15
Dependent Claim
44 words
The reagent of claim 10 for the production of superoxide wherein the autoxidizable redox partner is a substance which is able to induce a one-electron step and has a one-electron redox potential E o ' in the range of -150 mV to -500 mV.
Claim 16
Dependent Claim
34 words
The reagent of claim 11 for the production of superoxide wherein the redox partner is a bipyridilium salt, a triazonium salt, an anthraquinone derivative, a nitrofuran derivative, a ferredoxin, a pteridine or an isoalloxazine.
Claim 17
Dependent Claim
46 words
The reagent of claim 10 for the production of hydrogen peroxide wherein the autoxidizable redox partner is a substance which is able to induce a two-electron step and has a two-electron redox potential E o in the range of from 0 to mV to 150 mV.
Claim 18
Dependent Claim
19 words
The reagent of claim 11 for the production of hydrogen peroxide wherein the redox partner is a quinone derivative.
Claim 19
Dependent Claim
49 words
The reagent of claim 10 for the production of hydroxyl radicals wherein the autoxidizable redox partner is a substance which is able to induce a one-electron step and has a one-electron redox potential E' in the range of from -150 mV to -500 mV and additionally contains superoxide dismutase.
Claim 20
Independent Claim
81 words
A method for the determination of superoxide dismutase comprising the steps of coupling a superoxide-yielding reaction with two competing superoxide-consuming reactions, one superoxide-consuming reaction being an indicator reaction and the other superoxide-consuming reaction being a superoxide dismutase reaction, wherein, as superoxide-yielding reaction, there is used the reduction of oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase, of an autoxidizable one-electron step-inducing redox partner with a one-electron redox potential in the range of from -150 mV to -500 mV.
Claim 21
Independent Claim
81 words
A method for the determination of superoxide dismutase comprising the steps of coupling a superoxide-yielding reaction with two competing superoxide-consuming reactions, one superoxide-consuming reaction being an indicator reaction and the other superoxide-consuming reaction being a superoxide dismutase reaction, wherein, as superoxide-yielding reaction, there is used the reduction of oxygen with NAD(P)H in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase, of an autoxidizable one-electron step-inducing redox partner with a one-electron redox potential in the range of from -150 mV to -500 mV.
Claim 22
Dependent Claim
17 words
The method of claim 17 wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is NADP-ferredoxin-(Cyt c)-oxidoreductase or diaphorase from microorganisms.
Claim 23
Dependent Claim
29 words
The method of claim 17 wherein the redox partner is a bipyridilium salt, a triazonium salt, an anthraquinone derivative, a nitrofuran derivative, a ferredoxin, a pteridine or an isoalloxazine.
Claim 24
Dependent Claim
48 words
A reagent for the determination of superoxide dismutase by the process of claim 17 comprising NAD(P)H, an NAD(P)H-dependent, non-autoxidizable diaphorase, an autoxidizable, one-electron step-inducing redox partner with a one-electron redox potential in the range of from -150 mV to -500 mV and the components for an indicator reaction.
Claim 25
Dependent Claim
17 words
Reagent of claim 20 wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is NADP-ferredoxin (Cyt c)-oxidoreductase or diaphorase from microorganisms.
Claim 26
Dependent Claim
29 words
The reagent of claim 20 wherein the redox partner is a bipyridilium salt, a triazonium salt, an anthraquinone derivative, a nitrofuran derivative, a ferredoxin, a pteridine or an isoalloxazine.
Claim 27
Dependent Claim
26 words
The reagent of claim 20 wherein the components for the indicator reaction are a hydroxylammonium salt and a test system for the detection of nitrite ions.
Claim 28
Independent Claim
67 words
A method for the determination of NAD(P)H and NAD(P)H-yielding reactions comprising reducing oxygen to hydrogen peroxide by NAD(P)H and determining the hydrogen peroxide, wherein the reduction of the oxygen by NAD(P)H is carried out in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase, of a two-electron redox partner with a two-electron redox potential in the range of from 0 mV to 150 mV and of a buffer system.
Claim 29
Independent Claim
67 words
A method for the determination of NAD(P)H and NAD(P)H-yielding reactions comprising reducing oxygen to hydrogen peroxide by NAD(P)H and determining the hydrogen peroxide, wherein the reduction of the oxygen by NAD(P)H is carried out in the presence of an NAD(P)H-dependent, non-autoxidizable diaphorase, of a two-electron redox partner with a two-electron redox potential in the range of from 0 mV to 150 mV and of a buffer system.
Claim 30
Dependent Claim
17 words
The method of claim 24 wherein the NAD(P)H-dependent, non-autoxidizable diaphorase, is NADP-ferredoxin-(Cyt c)-oxidoreductase or diaphorase from microorganisms.
Claim 31
Dependent Claim
14 words
The method of claim 24 wherein a quinone derivative is used as redox partner.
Claim 32
Dependent Claim
64 words
A reagent for the determination of NAD(P)H and of NAD(P)H-yielding reactions according to the method of claim 24 comprising, in the form of a solution, of a powder mixture, a reagent tablet or a lyophilisate: an NAD(P)H-dependent, non-autoxidizable diaphorase, and two-electron redox partner with a two-electron redox potential in the range of from 0 mV to 150 mV, and a hydrogen peroxide detection system.
Claim 33
Dependent Claim
15 words
The process of claim 1, wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is diaphorase from Clostridium kluyveri.
Claim 34
Dependent Claim
15 words
The reagent of claim 10 wherein the NAD(P)H-dependent, non-autoxidizable diaphorase is diaphorase from Clostridium kluyveri.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-12-15 | 1990-06-16 | 1990-12-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-12-15 | 1994-06-16 | 1994-12-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-12-15 | 1998-06-16 | 1998-12-16 | $7700.00 | Window Closed / Lapsed |