Continuous analyte monitoring electrode with crosslinked enzyme

1. An electrode comprising a conductive surface and a conjugate comprising at least one enzyme molecule and at least one conductive nanoparticle covalently bound to each other, wherein the conjugate is covalently bound to the electrode surface via the at least one enzyme molecule and wherein the nanoparticle is not directly covalently bound to the conductive surface.

2. The electrode of claim 1 , wherein the at least one enzyme molecule is an H2O2 generating and/or consuming enzyme molecule selected from the group consisting of a glucose oxidase (EC 1.1.3.4), a hexose oxidase (EC 1.1.3.5), an (S)-2 hydroxy acid oxidase (EC 1.1.3.15), a cholesterol oxidase (EC 1.1.3.6), a galactose oxidase (EC 1.1.3.9), an alcohol oxidase (EC 1.1.3.13), an L-glutamate oxidase (EC 1.4.3.11) and an L-aspartate oxidase (EC 1.4.3.16).

3. The electrode of claim 1 , wherein the covalent binding of the at least one enzyme molecule to the electrode surface occurs via a sulfur containing functional group selected from the group consisting of a sulfide group and a disulfide group, and/or wherein the covalent binding of the at least one enzyme molecule to a nanoparticle occurs via a sulfur containing functional group selected from the group consisting of a sulfide group and a disulfide group.

4. The electrode of claim 1 , wherein the at least one enzyme molecule has been modified to incorporate at least one functional group for covalent binding to the electrode surface and a nanoparticle, and wherein the at least one enzyme molecule has been modified at the amino terminus and/or at an amino side chain group.

5. The electrode of claim 4 , wherein the at least one enzyme molecule has been modified by reacting the at least one enzyme with a functionalizing reagent in a molar ratio of enzyme to functionalization reagent of about 1:1 to about 1:10.

6. The electrode of claim 1 , wherein the nanoparticles are metal nanoparticles selected from the group consisting of platinum, palladium, iridium, gold and silver nanoparticles.

7. The electrode of claim 1 , wherein the nanoparticles have an average size of about 1 to about 100 nm.

8. The electrode of claim 1 , wherein the conjugate has an average size of about 10 nm to about 300 nm.

9. The electrode of claim 1 , wherein the conjugate does not contain a redox mediator.

10. The electrode of claim 1 , wherein the electrode surface is a metal surface.

11. An electrochemical sensor for measuring the concentration of an analyte comprising at least one electrode according to claim 1 .

12. The sensor of claim 11 , wherein the enzyme molecule is a functionalized glucose oxidase, and the analyte is glucose.

13. The sensor of claim 11 for in vivo or in vitro use.

14. A method of manufacturing an electrode of claim 1 , comprising the steps:

(a) preparing a conjugate of at least one enzyme molecule and at least one nanoparticle under conditions wherein only the at least one enzyme molecule, but not the nanoparticle in the conjugate has free functional groups for covalently binding to a conductive surface of an electrode, and

(b) covalently binding said conjugate to the conductive surface of the electrode, wherein the binding exclusively occurs via free functional groups on the at least one enzyme molecule.

15. A method of measuring an analyte in a tissue and/or a body fluid comprising using the electrode of claim 1 .

16. A method of measuring an analyte in a tissue and/or a body fluid comprising using the electrochemical sensor of claim 11 .

17. An electrode comprising:

a conductive surface; and

a conjugate comprising at least one enzyme molecule and at least one nanoparticle covalently bound to each other,

wherein the at least one enzyme molecule, and not the nanoparticle, has free functional groups covalently bound to the conductive surface.