Subcutaneous glucose electrode
A small diameter flexible electrode designed for subcutaneous in vivo amperometric monitoring of glucose is described. The electrode is designed to allow “one-point” in vivo calibration, i.e., to have zero output current at zero glucose concentration, even in the presence of other electroreactive species of serum or blood. The electrode is preferably layered, with the layers serially deposited within a recess upon the tip of a polyamide insulated gold wire. A first glucose concentration-to-current transducing layer can be overcoated with an electrically insulating and glucose flux limiting layer (second layer) on which, optionally, an immobilized interference-eliminating horseradish peroxidase based film is deposited. An outer layer is preferably biocompatible.
1. A glucose sensor comprising:
a working electrode; and
a multilayer membrane complex comprising:
an analyte responsive sensing layer disposed on top and in electrical contact with the working electrode, the analyte responsive sensing layer comprising a glucose-specific enzyme, a mediator and a first polymer selected from the group consisting of poly(vinylimidazole) and poly(vinyl pyridine), wherein at least one of the glucose-specific enzyme and the mediator is chemically bonded to the first polymer; and
a diffusion-limiting layer comprising a crosslinked second polymer disposed on top and in contact with the analyte responsive sensing layer.
2. The glucose sensor of claim 1 , wherein at least one of the glucose-specific enzyme and the mediator is covalently bonded to the first polymer.
3. The glucose sensor of claim 2 , wherein at least one of the glucose-specific enzyme and the mediator forms one or more of carbon-carbon, carbon-nitrogen, or metal-carbon covalent bonds with the first polymer.
4. The glucose sensor of claim 2 , wherein at least one of the glucose-specific enzyme and the mediator is covalently bonded to the first polymer through a linker moiety.
5. The glucose sensor of claim 2 , wherein the mediator is covalently bonded to the first polymer through one or more ligands of the mediator.
6. The glucose sensor of claim 5 , wherein the one or more ligands comprises a substituted or unsubstituted heterocyclic nitrogen-containing moiety.
7. The glucose sensor of claim 5 , wherein one or more ligands of the mediator are substituents of the first polymer backbone.
8. The glucose sensor of claim 2 , wherein the glucose-specific enzyme is covalently bonded to the first polymer through one or more side chains of the enzyme.
9. The glucose sensor of claim 8 , wherein the one or more side chains of the glucose-specific enzyme is selected from the group consisting of substituted or unsubstituted amines, alcohols, thiols, phenols, imidazoles, indols, and carboxylic acids.
10. The glucose sensor of claim 9 , wherein one or more side chains of the glucose-specific enzyme are substituents of the first polymer backbone.
11. The glucose sensor of claim 1 , wherein the first polymer is crosslinked.
12. The glucose sensor of claim 11 , wherein at least one of the glucose-specific enzyme and the mediator is crosslinked with the first polymer.
13. The glucose sensor of claim 12 , wherein the mediator is crosslinked to the first polymer.
14. The glucose sensor of claim 11 , wherein at least one of the glucose-specific enzyme and the mediator is crosslinked with the first polymer through one or more crosslinking agents.
15. The glucose sensor of claim 11 , wherein one or more side chains of the glucose-specific enzyme are crosslinked to the first polymer.
16. The glucose sensor of claim 1 , wherein the first polymer forms a coordination complex with the mediator.
17. The glucose sensor of claim 16 , wherein one or more nitrogen-containing moieties of the first polymer forms a coordination complex with a metal of the mediator.
18. The glucose sensor of claim 16 , wherein the mediator is ionically bonded to the first polymer.
19. The glucose sensor of claim 18 , wherein the first polymer comprises at least one negatively charged moiety and the mediator comprises at least one positively charged moiety.
20. The glucose sensor of claim 19 , wherein the glucose-specific enzyme is glucose dehydrogenase (GDH) or glucose oxidase (GOx).
21. The glucose sensor of claim 1 , wherein the analyte-responsive enzyme further comprises an enzyme cofactor.
22. The glucose sensor of claim 21 , wherein the enzyme cofactor is flavin adenine dinucleotide.
23. The glucose sensor of claim 1 , wherein the mediator comprises ruthenium or osmium.
24. The glucose sensor of claim 1 , wherein the second polymer is formed in situ on top of the first polymer.
25. The glucose sensor of claim 1 , wherein the multilayer membrane complex further comprises a biocompatible layer disposed on top and in contact with the diffusion-limiting layer.
26. The glucose sensor of claim 25 , wherein the multilayer membrane complex further comprises an interferent-eliminating layer disposed on top of the biocompatible layer.
27. The glucose sensor of claim 1 , wherein the diffusion-limiting layer is bound to the analyte responsive sensing layer.
28. The glucose sensor of claim 1 , wherein the electrode is a gold or carbon electrode.
29. The glucose sensor of claim 1 , wherein the sensor further comprises at least one other electrode, wherein the at least one other electrode is selected from the group consisting of a reference electrode, a counter electrode and a combined reference/counter electrode.
30. The glucose sensor of claim 1 , wherein the sensor has substantially no signal output when the concentration of glucose is zero.