Patent US5443701A successfully refreshed from Google Patents.
US5443701A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Electrobiochemical analytical method and electrodes
Assignee: Yissum Research Development Co of Hebrew University of Jerusalem
•
Filing: 1993-08-23
•
Granted: 1995-08-22
•
Expires: 2013-08-23
Abstract
Abstract An analytical method for determining the presence or concentration of an analyte in a liquid medium is provided in which electrons are transferred from an electrode material of an electrode to the redox center of an enzyme, by the mediation of an electron mediator, whereby in the presence of an analyte the enzyme calalyzed a redox reaction in which the analyte is converted into a product. By measuring either the concentration or the product or the charge which flows in this process, the presence and/or concentration of the analyte in the medium is determined. Either or both of the enzyme and the electron mediator are immobilized on the electrode.
Inventors
- 👤 Itamar Willner
- 👤 Evgeny Katz
- 👤 Azalia Riklin
- 👤 Ron Kasher
- 👤 Benjamin Shoham
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 10:49
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/001; C12Q1/004; C12Q1/002; C12Q1/003; Y; Y10; Y10S; Y10S435/00; Y10S435/817
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
128 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an electrobiochemical system, the electrobiochemical system comprising a test electrode which has chemisorbed sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte.
Claim 2
Dependent Claim
128 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an electrobiochemical system, the electrobiochemical system comprising a test electrode which has chemisorbed sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte.
Claim 3
Dependent Claim
17 words
A method according to claim 1 wherein the electron mediator material is immobilized on the test electrode.
Claim 4
Dependent Claim
43 words
The analytical method of claim 1 wherein the method further includes the step of selecting an electron mediator wherein the electron mediator is selected from the group consisting of viologen, pyridinium, acridine, ferrocene, phenothiazine, substituted pyridinium, substituted acridine, substituted phenothiazine and substituted ferrocene.
Claim 5
Dependent Claim
52 words
The analytical method of claim 3 wherein the method further includes the step of selecting an electron mediator from the group consisting of compounds having the following formulae (IV) to (VIII): ##STR14## wherein n is an integer between 1 and 16; ##STR15## wherein Y is CO 2 H or NH 2 ##STR16##
Claim 6
Dependent Claim
104 words
The analytical method of claim 1 wherein the method further includes the step of selecting an electron mediator having the general formulae (IX): U--R.sup.4 --X.sup.3 (IX) wherein U is a member selected from the group consisting of viologen, pyridinium, acridine, ferrocene, phenothiazine, substituted viologen, substituted pyridinium, substituted acridine, substituted ferrocene and substituted phenothiazine, R 4 is a connecting group, and X 3 is a functional group, which forms a covalent bond with a moiety of said at least one component and reacting the electron mediator with the moiety of at least one component to bond the election mediator in vicinity of the redox enzyme.
Claim 7
Dependent Claim
45 words
An analytical method according to claim 1 wherein the method further includes the steps of: selecting an enzyme; selecting a bridging group; reacting the bridging group with the enzyme; and reacting the enzyme with already bound enzyme through bridging groups to form a test electrode.
Claim 8
Dependent Claim
53 words
The method of claim 6 wherein the bridging group have the following formula (X): W.sup.1 --R.sup.b --W.sup.2 (X) wherein W 1 and W 2 may be the same or different from one another and form a covalent bond with a moiety of at least one component and R b represents a connecting group.
Claim 9
Dependent Claim
46 words
The method of claim 6 wherein the bridging group is selected from the group consisting of proteins, polypeptides and polymers, the proteins, polypeptides and polymers having at least one functional group X 2 which forms a covalent bond with a moiety of at least one component.
Claim 10
Dependent Claim
30 words
The method of claim 6 wherein the enzyme molecules selected for the external layer are non-redox enzymes, the non-redox enzymes catalyzing the reaction of interfering material in the liquid medium.
Claim 11
Dependent Claim
93 words
An electrode comprising an electrode material has chemisorbed sulphur containing moieties, the electrode having immobilized thereon a plurality of complexes, each complex comprising a linking group having a sulphur containing moiety and at least one, of a redox enzyme component or a molecular electron mediator component, all components of the complex are covalently bound to one another, the electron mediator transfers electrons between the electrode material and the redox center of the redox enzyme whereby, in the presence of analyte, the enzyme catalyzes a reaction in which the analyte is convened to product.
Claim 12
Dependent Claim
93 words
An electrode comprising an electrode material has chemisorbed sulphur containing moieties, the electrode having immobilized thereon a plurality of complexes, each complex comprising a linking group having a sulphur containing moiety and at least one, of a redox enzyme component or a molecular electron mediator component, all components of the complex are covalently bound to one another, the electron mediator transfers electrons between the electrode material and the redox center of the redox enzyme whereby, in the presence of analyte, the enzyme catalyzes a reaction in which the analyte is convened to product.
Claim 13
Dependent Claim
99 words
The electrode of claim 10, wherein the linking group has the following general formula (I); Z--R.sup.1 --Q wherein: Z represents a sulphur containing moiety, Q represents a group selected from the group consisting of X 1 and P wherein X 1 is a functional group which forms a covalent bond with a moiety of at least one component and P is selected from the group consisting of proteins, polypeptides and polymers, each having at least one functional group X 2 which forms a covalent bond with a moiety of at least one component, R 1 represents a connecting group.
Claim 14
Dependent Claim
31 words
An electrode according to claim 11, wherein X 1 or X 2 is selected from the group consisting of an amine group, a carboxyl group, -N=C=S,--N=C--O, an acyl group and ##STR17##
Claim 15
Dependent Claim
97 words
An electrode according to claim 12 wherein the acyl group has the formula --R a --CO--G wherein G is selected from the group consisting of OH, halogen, and OR b wherein R a and R b being independently selected from the group consisting of a C 1 -C 12 alkyl chain, C 1 -C 12 alkenyl chain, C 1 -C 12 alkynyl chain, a phenyl containing chain, a substituted C 1 -C 12 alkyl chain, a substituted C 1 -C 12 alkenyl chain, a substituted C 1 -C 12 alkynyl chain and a substituted phenyl containing chain.
Claim 16
Dependent Claim
32 words
An electrode according to any one of claims 11 to 13, comprising linking groups of the general formula (I), wherein P is polyamine or a polypeptide with free amine or carboxyl groups.
Claim 17
Dependent Claim
19 words
An electrode according to claim 14 wherein P is selected from the group consisting of polylysine, polyglutamate and polyethyleneimine.
Claim 18
Dependent Claim
36 words
An electrode according to claim 14, wherein P carries at least one functional group X 1 wherein X 1 is a functional group which forms a covalent bond with a moiety of at least one component.
Claim 19
Dependent Claim
29 words
An electrode according to claim 11, wherein R 1 is selected from the group consisting of a covalent bond, alkylene chains, alkenylene chains, alkynylene chains, and phenyl containing chains.
Claim 20
Dependent Claim
110 words
An electrode according to claim 17 wherein R 1 is selected from the group consisting of a chemical bond, and a group having the following formula (II a), (II b) and (III): wherein: R 2 and R 3 may be the same or different and are selected from the group consisting of a covalent bond and straight or branched alkylene, alkenylenes, alkynylene molecules having 1-12 carbon atoms; A and B may be the same or different and are selected from the group consisting of oxygen and sulfur; Ph is a phenyl which is selected from the group consisting of phenyl, alkylphenyl, polyalkylphenyl, phenyl sulfate, alkylphenyl sulfate, polyalkylphenyl sulfate, polyalkylphenyl polysulfate.
Claim 21
Dependent Claim
40 words
An electrode according to anyone of claims 10-18, wherein the electron mediators have a mediator moiety U being a member selected from the group consisting of viologen, pyridinium, acridine, ferrocene, phenothiazine, substituted pyridinium, substituted acridine, substituted phenothiazine and substituted ferrocene.
Claim 22
Dependent Claim
46 words
An electrode according to claim 19, wherein U is a member selected from the group consisting of compounds having the following formulae (IV) to (VIII): ##STR18## wherein n is an integer between 1 and 16; ##STR19## wherein Y is CO 2 H or NH 2 ##STR20##
Claim 23
Dependent Claim
81 words
An electrode according to any one of claims 10-18 wherein the electron mediator group forms part of said complex and has the general formula (IX): U--R.sup.4 --X.sup.3 (IX) wherein U is a member selected from the group consisting of viologen, pyridinium, acridine, ferrocene, phenothiazine, substituted viologen, substituted pyridinium, substituted acridine, substituted ferrocene and substituted phenothiazine, R 4 is a connecting group, and X 3 is a functional group, which forms a covalent bond with a moiety of at least one component.
Claim 24
Dependent Claim
28 words
An electrode according to claim 10 comprising several layers of enzymes, enzymes of one layer being linked to enzymes of an adjacent layer by means of bridging groups.
Claim 25
Dependent Claim
53 words
An electrode according to claim 22, wherein the bridging groups have the following formula (X): W.sup.1--R.sup.6 --W.sup.2 (X) wherein W 1 and W 2 may be the same of different from one another and forms a covalent bond with a moiety of at least one component and R 6 represents a connecting group.
Claim 26
Dependent Claim
47 words
An electrode according to claim 22, wherein the bridging group is selected from the group consisting of proteins, polypeptides and polymers, the proteins, polypeptides and polymers having at least one functional group X 2 which forms a covalent bond with a moiety of at least one component.
Claim 27
Dependent Claim
32 words
An electrode according to any one of claims 22 to 24 wherein the enzyme molecules in the external layer are non-redox enzymes which catalyst the reaction of agents other than the analyte.
Claim 28
Dependent Claim
67 words
A process for preparing an electrode comprising the steps: selecting a sulfur chemisorption electrode material; selecting a complex comprising a linking group having a sulphur containing moiety, chemisorption of the sulfur containing moiety onto the electrode material; selecting at least one of a redox enzyme and an electron mediator and reacting at least one of the redox enzyme and an electron mediator with the chemisorbed linking group.
Claim 29
Dependent Claim
67 words
A process for preparing an electrode comprising the steps: selecting a sulfur chemisorption electrode material; selecting a complex comprising a linking group having a sulphur containing moiety, chemisorption of the sulfur containing moiety onto the electrode material; selecting at least one of a redox enzyme and an electron mediator and reacting at least one of the redox enzyme and an electron mediator with the chemisorbed linking group.
Claim 30
Dependent Claim
66 words
A process for preparing an electrode comprising the steps: selecting a sulfur chemisorption electrode material, selecting a sulfur containing linking group; selecting at least one of a redox enzyme and an electron mediator; reacting the selected linking group with at least one of the selected redox enzymes and electron mediators to form a complex; and reacting the complex with the electrode material to form the electrode.
Claim 31
Dependent Claim
66 words
A process for preparing an electrode comprising the steps: selecting a sulfur chemisorption electrode material, selecting a sulfur containing linking group; selecting at least one of a redox enzyme and an electron mediator; reacting the selected linking group with at least one of the selected redox enzymes and electron mediators to form a complex; and reacting the complex with the electrode material to form the electrode.
Claim 32
Dependent Claim
29 words
A process according to claims 26 or 27, further comprising: (a) unfolding the enzyme molecule; (b) binding electron mediator groups to the unfolded enzyme molecule; (c) refolding the enzyme.
Claim 33
Dependent Claim
46 words
The analytical method of claim 1 wherein the method further includes the steps of selecting a sulfur containing linking group; binding the sulfur containing linking group to the redox enzyme to form a linking group-enzyme component; and binding the linking group-enzyme component to the electrode material.
Claim 34
Dependent Claim
110 words
The analytical method of claim 29 wherein the method further includes the step of selecting a linking group with the following general formula (I): Z--R.sup.1 --Q (I) wherein: Z represents a sulfur containing moiety, Q represents a group selected from the group consisting of X 1 and P wherein X 1 is a functional group which forms a covalent bond with a moiety of at least one component and P is selected from the group consisting of proteins, polypeptides and polymers, each having at least one functional group X 2 which forms a covalent bond with a moiety of at least one component and R 1 represents a connecting group.
Claim 35
Dependent Claim
48 words
The analytical method of claim 30 wherein the method further includes the step of selecting the X 1 or X 2 linking group wherein X 1 and X 2 are independently selected from the group consisting of an amine group, a carboxyl group, --N=C=S,--N=C=O, aryl group and ##STR21##
Claim 36
Dependent Claim
73 words
The analytical method of claim 31, wherein the method further includes the step of selecting an acyl group which has the formula --R a --CO--G, wherein G is selected from the group consisting of OH, halogen and OR b , and R a or R b being independently selected from the group consisting of C 1 -C 12 alkyl, C 1 -C 12 alkynyl, C 1- C 12 alkenyl and phenyl containing chains.
Claim 37
Dependent Claim
32 words
The analytical method of claim 30 wherein the method further includes the step of selecting a group P from the group consisting of polyamides and polypeptides with free amine or carboxyl groups.
Claim 38
Dependent Claim
27 words
The analytical method of claim 33 wherein the method further includes the step of selecting a group P from the group consisting of polylysine, polyglutamate and polyethyleneimine.
Claim 39
Dependent Claim
35 words
The analytical method claim 33 wherein P carries a plurality of functional groups X 1 wherein X 1 is a functional group which forms a covalent bond with a moiety of at least one component.
Claim 40
Dependent Claim
29 words
The analytical method of claim 30 wherein R 1 is selected from the group consisting of a covalent bond, alkylene chain, alkenyl chain, alkynyl chain and phenyl containing chain.
Claim 41
Dependent Claim
113 words
The analytical method of claim 30 wherein R 1 is selected from the group consisting of a chemical bond, and a group having the following formula (II a), (II b) and (III): ##STR22## wherein: R 2 and R 3 may be the same or different and are selected from the group consisting of a covalent bond and straight or branched alkylene, alkenylenes, alkynylene molecules having 1-12 carbon atoms; A and B may be the same or different and are independently selected from the group consisting of oxygen and sulfur; and Ph is a phenyl which is selected from the group consisting of phenyl, alkylphenyl, polyalkylphenyl, phenyl sulfate, alkylphenyl sulfate, polyalkylphenyl sulfate, polyalkylphenyl polysulfate.
Claim 42
Dependent Claim
122 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte.
Claim 43
Dependent Claim
122 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte.
Claim 44
Dependent Claim
45 words
The method of claim 38 wherein the method further includes the steps of selecting an electron mediator group; reacting said electron mediator group with the redox enzyme to form a redox enzyme-electron mediator group; and introducing the redox enzyme-electron mediator complex into the liquid medium.
Claim 45
Dependent Claim
18 words
The method of claim 39 wherein the analyte concentration is measured by measuring the flow of electric charge.
Claim 46
Dependent Claim
24 words
The method of claim 39 wherein the analyte concentration is measured by measuring the concentration of the redox reaction product in the liquid medium.
Claim 47
Dependent Claim
129 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the flow of electric charge.
Claim 48
Dependent Claim
129 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the flow of electric charge.
Claim 49
Dependent Claim
135 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the concentration of the redox reaction product in the liquid medium.
Claim 50
Dependent Claim
135 words
An analytical method for determining the presence or concentration of analyte in a liquid medium comprising the steps of; providing an electrobiochemical system comprising a test electrode which has a sulfur containing moiety; providing an electron mediator; chemisorbing the election mediator onto the test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; providing a redox enzyme for oxidizing or reducing the analyte and for interacting with the electron mediator; introducing the redox enzyme into the liquid medium; applying a constant or alternate electric potential to the test electrode so that electrons pass between the mediator group, test electrodes and redox enzyme; allowing the enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the concentration of the redox reaction product in the liquid medium.
Claim 51
Dependent Claim
138 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an appropriate electrobiochemical system, the electrobiochemical system comprising selecting a test electrode which has chemisorption of sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the flow of electron charge.
Claim 52
Dependent Claim
138 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an appropriate electrobiochemical system, the electrobiochemical system comprising selecting a test electrode which has chemisorption of sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the flow of electron charge.
Claim 53
Dependent Claim
144 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an appropriate electrobiochemical system, the electrobiochemical system comprising selecting a test electrode which has chemisorption of sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the concentration of the redox reaction product in the liquid medium.
Claim 54
Dependent Claim
144 words
An analytical method for determining the presence or concentration of an analyte in a liquid medium comprising the steps of: selecting an appropriate electrobiochemical system, the electrobiochemical system comprising selecting a test electrode which has chemisorption of sulfur containing moieties; selecting a redox enzyme, the redox enzyme selectively converts the analyte to a product; binding the redox enzyme to the test electrode; selecting an electron mediator material which transfers electrons to and from the redox enzyme and test electrode; introducing a liquid medium with an analyte to the electrobiochemical system; applying a constant or alternating electric potential to the test electrode so that electrons pass between the redox enzyme and the test electrode allowing the redox enzyme to catalyze the reaction of the analyte; and measuring the concentration of the analyte by measuring the concentration of the redox reaction product in the liquid medium.
References Cited (Backward Citations)
US4964972A
Cited
US5232574A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1998-08-21 | 1999-02-20 | 1999-08-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-08-21 | 2003-02-20 | 2003-08-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-08-21 | 2007-02-20 | 2007-08-22 | $7700.00 | Window Closed / Lapsed |