Patent US4797181A successfully refreshed from Google Patents.
US4797181A
Expired / Lapsed
US
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Flavin cofactor modified electrodes and methods of synthesis and use
Assignee: Verizon Laboratories Inc
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Filing: 1987-08-03
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Granted: 1989-01-10
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Expires: 2007-08-03
Abstract
Abstract An electrochemically coupled cofactor modified electrode for mediating the heterogeneous or homogeneous catalysis of a biochemical reaction by an enzyme. The electrode coating comprises mercapto-flavin derivative bonded to the electrode surface. Methods for preparation and use of the modified electrode are also presented.
Inventors
- 👤 Charles N. Durfor
- 👤 Mark L. Bowers
- 👤 Barbara A. Yenser
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 16:34
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/001; C12Q1/004; Y; Y10; Y10S; Y10S435/00; Y10S435/817
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
93 words
A cofactor modified electrode comprising: an electrode substrate having a working surface of an electrically conducting or semiconducting material having an outer oxide or hydroxyl containing layer; ad a redox active material electrochemically bonded to the outer layer by means of a mercaptosilane linking group; wherein the redox active material is derived from a redox active flavin compound having the formula ##STR5## wherein R 1 is a ribose derivative, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alphamethyl position on the redox active material.
Claim 2
Dependent Claim
93 words
A cofactor modified electrode comprising: an electrode substrate having a working surface of an electrically conducting or semiconducting material having an outer oxide or hydroxyl containing layer; ad a redox active material electrochemically bonded to the outer layer by means of a mercaptosilane linking group; wherein the redox active material is derived from a redox active flavin compound having the formula ##STR5## wherein R 1 is a ribose derivative, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alphamethyl position on the redox active material.
Claim 3
Dependent Claim
46 words
An electrode according to claim 1 wherein the redox active material is criented to permit coupling of the redox active material to a suitable apoenzyme in a solution to form a renatured enzyme and to permit electron transfer between the electrode surface and the renatured enzyme.
Claim 4
Dependent Claim
32 words
A electrode according to claim 1 wherein the redox active material is oriented to permit electron transfer between the redox active material and a holoenzyme in a solution to activate the holoenzyme.
Claim 5
Dependent Claim
46 words
An electrode according to claim 1 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR6##
Claim 6
Dependent Claim
35 words
An electrode according to claim 1 wherein the working surface of the electrode is of a material selected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon, and conducting polymers.
Claim 7
Dependent Claim
53 words
An electrode according to claim 1 wherein the linking group is a derivative of a silane compound having the formula ##STR7## wherein R 4 , R 5 , and R 6 are independently selected from the Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 8
Dependent Claim
156 words
A cofactor modified electrode comprising: an electrode substrate having a working surface of an electrically conducting or semiconducting material selected from the group consisting of platinum, gold, silicon, InP, graphite, glassy carbon, tin dioxide, and conducting polymers, the working surface having an outer oxide or hydroxyl layer; and a redox active coating electrochemically bonded to the outer layer and comprising groups of the formula ##STR8## wherein R 1 is a ribose derivative selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and --CH 2 (CHOH) 3 CH 2 O--P--O--P--O--CH 2 ##STR9## and R 2 and R 3 are each an --O--C 2 H 5 group or an --O-- link to an adjacent silane group or to the outer layer of the electrode working surface, and may be the same or different.
Claim 9
Dependent Claim
156 words
A cofactor modified electrode comprising: an electrode substrate having a working surface of an electrically conducting or semiconducting material selected from the group consisting of platinum, gold, silicon, InP, graphite, glassy carbon, tin dioxide, and conducting polymers, the working surface having an outer oxide or hydroxyl layer; and a redox active coating electrochemically bonded to the outer layer and comprising groups of the formula ##STR8## wherein R 1 is a ribose derivative selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and --CH 2 (CHOH) 3 CH 2 O--P--O--P--O--CH 2 ##STR9## and R 2 and R 3 are each an --O--C 2 H 5 group or an --O-- link to an adjacent silane group or to the outer layer of the electrode working surface, and may be the same or different.
Claim 10
Independent Claim
172 words
A method of effecting electron transfer between an electrode having a working surface of an electrically conducting or semiconducting material and an enzyme comprising the steps of: modifying the working surface of the electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin -compound having the formula ##STR10## wherein the redox active material is selected to activate a protein moiety of the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; immersing the modified working surface of the electrode in an electrolyte solution containing the protein moiety of the enzyme; and applying an electric potential to the electrode sufficient to effect the electron transfer between the electrode surface and the enzyme through the redox active material.
Claim 11
Independent Claim
172 words
A method of effecting electron transfer between an electrode having a working surface of an electrically conducting or semiconducting material and an enzyme comprising the steps of: modifying the working surface of the electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin -compound having the formula ##STR10## wherein the redox active material is selected to activate a protein moiety of the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; immersing the modified working surface of the electrode in an electrolyte solution containing the protein moiety of the enzyme; and applying an electric potential to the electrode sufficient to effect the electron transfer between the electrode surface and the enzyme through the redox active material.
Claim 12
Dependent Claim
120 words
A method according to claim 8 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR11## the working surface of the electrode is of a material sclected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon, and conducting polymers, and the linking group is a derivative of a silane compound having the formula ##STR12## wherein R 4 , R 5 and R 6 are independently selected from the Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 13
Independent Claim
172 words
A method of effecting electron transfer between an electrode having a working surface of an electrically conducting or semiconducting material and an enzyme comprising the steps of: modifying the working surface of the electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group a redox active material derived from a redox active flavin compound having the formula ##STR13## wherein the redox active material is selected to activate the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material immersing the modified working surface of the electrode in an electrolyte solution in an electrochemical cell; introducing the enzyme to the electrochemical cell; and applying an electric potential to the electrode sufficient to effect the electron transfer between the electrode surface and the enzyme through the redox active material.
Claim 14
Independent Claim
172 words
A method of effecting electron transfer between an electrode having a working surface of an electrically conducting or semiconducting material and an enzyme comprising the steps of: modifying the working surface of the electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group a redox active material derived from a redox active flavin compound having the formula ##STR13## wherein the redox active material is selected to activate the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material immersing the modified working surface of the electrode in an electrolyte solution in an electrochemical cell; introducing the enzyme to the electrochemical cell; and applying an electric potential to the electrode sufficient to effect the electron transfer between the electrode surface and the enzyme through the redox active material.
Claim 15
Dependent Claim
25 words
A method according to claim 10 wherein the step of introducing the enzyme to the electrochemical cell comprises adding the enzyme to the electrolyte solution.
Claim 16
Dependent Claim
28 words
A method according to claim 10 wherein the step of introducing the enzyme to the electrochemical cell comprises immobilizing the enzyme on the working surface of the electrode.
Claim 17
Dependent Claim
120 words
A method according to claim 10 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR14## the working surface of the electrode is of a material selected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon, and conducting polymers, and the linking group is a derivative of a silane compound having the formula ##STR15## wherein R 4 , R 5 and R 6 are independently selected from the Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 18
Independent Claim
187 words
A method of electrochemically mediating the catalysis of a reaction involving one or more biochemical substrates by an enzyme comprising the steps of: modifying a working surface of a working electrode by electrochemically bonding to an existing or derivatized oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin compound having the formula ##STR16## wherein the redox active material is selected H to activate a protein moiety of the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; establishing in an electrochemical cell having at least the modified electrode and an auxiliary electrode, an electrolyte solution containing the biochemical substrate(s) and the protein moiety of the enzyme suitable for catalyzing the reaction; and applying an electric potential to the electrodes sufficient to effect electron transfer between the modified electrode and the biochemical substrate through the redox active material and the enzyme.
Claim 19
Independent Claim
187 words
A method of electrochemically mediating the catalysis of a reaction involving one or more biochemical substrates by an enzyme comprising the steps of: modifying a working surface of a working electrode by electrochemically bonding to an existing or derivatized oxide or hydroxyl layer on the working surface, by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin compound having the formula ##STR16## wherein the redox active material is selected H to activate a protein moiety of the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; establishing in an electrochemical cell having at least the modified electrode and an auxiliary electrode, an electrolyte solution containing the biochemical substrate(s) and the protein moiety of the enzyme suitable for catalyzing the reaction; and applying an electric potential to the electrodes sufficient to effect electron transfer between the modified electrode and the biochemical substrate through the redox active material and the enzyme.
Claim 20
Dependent Claim
39 words
A method according to claim 14 wherein the enzyme is a metalloenzyme and the electrolyte solution further contains one or more metallic ions suitable for coupling with the protein moiety and the redox active material to complete the enzyme.
Claim 21
Dependent Claim
120 words
A method according to claim 14 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR17## the working surface of the electrode is of a material selected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon, and conducting polymers, and the linking group is a derivative of a silane compound having the formula ##STR18## wherein R 4 , R 5 and R 6 are independently selected from the Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 22
Independent Claim
182 words
A method of electrochemically mediating the catalysis of a reaction involving one or more biochemical substrates by an enzyme comprising the steps of: modifying a working surface of an electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin compound having the formula ##STR19## wherein the redox active ,aterial is selected to activate the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; establishing in an electrochemical cell having at least the modified electrode and an auxiliary electrode an electrolyte solution containing the biochemical substrate(s) introducing to the electrochemical cell the enzyme suitable for catalyzing the reaction; and applying an electric potential to the electrodes sufficient to effect electron transfer between the modified electrode and the biochemical substrate through the redox active material and the enzyme.
Claim 23
Independent Claim
182 words
A method of electrochemically mediating the catalysis of a reaction involving one or more biochemical substrates by an enzyme comprising the steps of: modifying a working surface of an electrode by electrochemically bonding to an existing or derivatized outer oxide or hydroxyl layer on the working surface by means of a mercaptosilane linking group, a redox active material derived from a redox active flavin compound having the formula ##STR19## wherein the redox active ,aterial is selected to activate the enzyme, R 1 being a ribose derivative selected to provide the selected redox active material, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material; establishing in an electrochemical cell having at least the modified electrode and an auxiliary electrode an electrolyte solution containing the biochemical substrate(s) introducing to the electrochemical cell the enzyme suitable for catalyzing the reaction; and applying an electric potential to the electrodes sufficient to effect electron transfer between the modified electrode and the biochemical substrate through the redox active material and the enzyme.
Claim 24
Dependent Claim
25 words
A method according to claim 17 wherein the step of introducing the enzyme to the electrochemical cell comprises adding the enzyme to the electrolyte solution.
Claim 25
Dependent Claim
28 words
A method according to claim 17 wherein the step of introducing the enzyme to the electrochemical cell comprises immobilizing the enzyme on the working surface of the electrode.
Claim 26
Dependent Claim
120 words
A method according to claim 17 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR20## the working surface of the electrode is of a material selected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon, and conducting polymers, and the linking group is a derivative of a silane compound having the formula ##STR21## wherein R 4 , R 5 and R 6 are independently selected from the Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 27
Independent Claim
92 words
A method for coating an electrode substrate having a working surface of an electrically conductive or semiconductive material having an outer oxide or hydroxyl layer comprising: electrochemically bonding a redox active material to the outer layer by means of a mercaptosilane linking group; wherein the redox active material is derived from a redox active flavin compound having the formula ##STR22## wherein R 1 is a ribose derivative, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material.
Claim 28
Independent Claim
92 words
A method for coating an electrode substrate having a working surface of an electrically conductive or semiconductive material having an outer oxide or hydroxyl layer comprising: electrochemically bonding a redox active material to the outer layer by means of a mercaptosilane linking group; wherein the redox active material is derived from a redox active flavin compound having the formula ##STR22## wherein R 1 is a ribose derivative, and the redox active material is bonded to the sulfur atom of the linking group through the 8-alpha methyl position on the redox active material.
Claim 29
Dependent Claim
46 words
A method according to claim 21 wherein R 1 is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR23##
Claim 30
Independent Claim
162 words
A method of preparing a working electrode for electrochemically mediating the catalysis of a biochemical reaction by an enzyme comprising the steps of: immersing a working surface of the electrode, the working surface being of an electrically conducting or semiconducting material having an outer oxide layer on the working surface, in a solution containing a mercaptosilane compound from which a suitable linking group may be derived, for a sufficient time to electrochemically bond a sufficient quantity of a derivative of the mercaptosilane to the outer oxide layer to support sufficient redox active material; immersing the mercaptosilane derivatized working surface in a solution containing a redox active material derived from a redox active flavin compound having the formula ##STR24## wherein R 1 is a ribose derivative, for a sufficient time to electrochemically bond sufficient redox active material to the outer oxide layer by means of the linking group to provide a working electrode caable of electrochemically mediating the catalysis of the biochemical reaction.
Claim 31
Independent Claim
162 words
A method of preparing a working electrode for electrochemically mediating the catalysis of a biochemical reaction by an enzyme comprising the steps of: immersing a working surface of the electrode, the working surface being of an electrically conducting or semiconducting material having an outer oxide layer on the working surface, in a solution containing a mercaptosilane compound from which a suitable linking group may be derived, for a sufficient time to electrochemically bond a sufficient quantity of a derivative of the mercaptosilane to the outer oxide layer to support sufficient redox active material; immersing the mercaptosilane derivatized working surface in a solution containing a redox active material derived from a redox active flavin compound having the formula ##STR24## wherein R 1 is a ribose derivative, for a sufficient time to electrochemically bond sufficient redox active material to the outer oxide layer by means of the linking group to provide a working electrode caable of electrochemically mediating the catalysis of the biochemical reaction.
Claim 32
Dependent Claim
130 words
A method according to claim 23 wherein the working surface of the electrode is of a material selected from the group consisting of platinum, gold, tin dioxide, silicon, InP, graphite, glassy carbon and conducting polymers; the ribose derivative is selected from the group consisting of --CH 2 (CHOH) 3 --CH 2 OH, --CH 2 (CHOH) 3 CH 2 O--PO 3 2-- , --CH 2 (CHOCOCH 3 ) 3 CH 2 OCOCH 3 , and ##STR25## and the compound from which a suitable linking group may be derived is a silane compound having the formula ##STR26## wherein R 4 , R 5 , and R 6 are independently selected from the group consisting of Si ligands displaceable by a hydroxide anion, and n is a positive integer from 1 to 20.
Claim 33
Dependent Claim
72 words
A method according to claim 23 wherein the immersing steps take place in an electrochemical cell containing an auxiliary electrode, and further comprising the step of establishing an electrical potential across the electrodes during bond formation of sufficient density and for sufficient time to electrochemically bond sufficient linking groups and redox active material to the outer layer to provide a working electrode capable of electrochemically mediating the catalysis of the biochemical reaction.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-01-10 | 1992-07-11 | 1993-01-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-01-10 | 1996-07-11 | 1997-01-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-01-10 | 2000-07-11 | 2001-01-10 | $7700.00 | Window Closed / Lapsed |