Biosensor carrying redox enzymes
First Claim
Patent Images
1. An electrode carrying immobilized groups having the general formulaV—
- W—
X—
Y—
Zwherein V is a binding moiety that can chemically associate with, attach to, or chemically sorb onto the electrode;
W is an electron mediator group that can transfer electrons between the electrode and Y;
X is a linker moiety;
Y is a cofactor of a redox enzyme having, when not bound to X, at least one pair of cis hydroxyl groups; and
Z is a redox enzyme;
characterized in that X is a boromic acid derivative.
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Abstract
The present invention concerns an electrode carrying immobilized redox enzymes such that electric charge can flow between an electron mediator group to the enzyme cofactor by the use of boronic acid or a boronic acid derivative that acts as a linker moiety between the cofactor and the electron mediator group. The invention also concerns devices and systems that make use of the electrode of the invention, such as bio-sensors and fuel cells, the electrode being one of the components thereof.
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Citations
19 Claims
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1. An electrode carrying immobilized groups having the general formula
V— - W—
X—
Y—
Zwherein V is a binding moiety that can chemically associate with, attach to, or chemically sorb onto the electrode;
W is an electron mediator group that can transfer electrons between the electrode and Y;
X is a linker moiety;
Y is a cofactor of a redox enzyme having, when not bound to X, at least one pair of cis hydroxyl groups; and
Z is a redox enzyme;
characterized in that X is a boromic acid derivative. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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3. An electrode according to claim 1, wherein X is aminophenyl boronic acid derivative.
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4. An electrode according to claim 1, wherein the cofactor is selected from FAD, NAD+ and NADP+.
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5. An electrode according to claim 1, wherein the enzyme is selected from glucose oxidase, lactate dehydrogenase and malic enzyme.
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6. An electrode according to claim 1 wherein the enzyme is crosslinked using a bifunctional cross-inker capable to react with amino groups, to improve its binding to the electrode.
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7. An electrode according to claim 6, wherein said cross-inker is glutaric dialdehyde.
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8. A device comprising an electrode according to claim 1.
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9. A device according to claim 8, being a biosensor.
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10. A biosensor according to claim 9, comprising an electronic circuitry for energizing the electrode and measuring the response.
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11. A device according to claim 8, being a fuel cell.
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12. A fuel cell according to claim 1, comprising a substrate for the redox enzyme.
- W—
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13. A process for preparing an electrode carrying immobilized redox enzymes Z comprising:
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(i) forming a layer on the surface of the electrode comprising groups of the formula V—
W—
X—
Y, wherein V is a binding moiety that can chemically associate with, attach to, or chemically sorb onto the electrode and W is an electron mediator group that can transfer electrons between the electrode and Y, Y being a cofactor of the enzyme and X is a linker group;
(ii) contacting said electrode with one or more enzyme molecule, devoid of a cofactor to complex said enzymes with said cofactor to yield a immobilized groups V—
W—
X—
Y—
Z, wherein Z is a catalytically functional enzyme that can catalyze a redox reaction;
said method being characterized in thatsaid cofactor has at least one pair of cis hydroxyl groups and said process comprises binding said cofactor to said electron mediator group by a group X being a boronic acid derivative. - View Dependent Claims (15, 16, 19)
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14. A process for preparing an electrode carrying immobilized redox enzymes Z comprising:
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(i) forming a layer on the surface of the electrode comprising groups of the formula V—
W, wherein V is a binding moiety that can chemically associate with, attach to, or chemically sorb onto the electrode and W is an electron mediator group that can transfer electrons between the electrode and a cofactor Y of the enzyme;
(ii) binding Y to said groups; and
(iii) contacting said electrode with one or more enzyme molecule, devoid of a cofactor to complex said enzymes with said cofactor to yield functional enzymes that can catalyze a redox reaction;
said method being characterized in thatsaid cofactor has at least one pair of cis hydroxyl groups and said process comprises binding said cofactor to said electron mediator group by a group X being a boronic acid derivative. - View Dependent Claims (17, 18)
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Specification