Enzyme electrodes and improvements in the manufacture thereof

• US 5,160,418 A
• Filed: 07/18/1989
• Issued: 11/03/1992
• Est. Priority Date: 07/28/1988
• Status: Expired due to Term

• Current Assignee: Cambridge Life Sciences PLC
• Original Assignee: Cambridge Life Sciences PLC
• Inventors: Mullen, William H.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Bell, Bruce F.
• Application Number:
  US07/381,580
• Time in Patent Office: 1,204 Days
• Field of Search: 204/403, 204/418, 204/153.12, 204/153.2, 204/290 R, 204/291, 204/294, 435/817, 435/179
• US Class Current: 204/403.14
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: Y02E 60/50 Fuel cells
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

Enzyme electrodes are disclosed consisting essentially of a uniform homogeneous layer of a finely divided platinum group metal or oxide, preferably preadsorbed onto the surface of an activated carbon or grapite powder, and deposited from suspension upon the surface of an electrically conductive substrate and in admixture with an enzyme and optionally a water soluble or water-dispersible binder. Preferably the enzyme electrodes are produced by coating the substrate with a suspension of the enzyme, the finely divided platinum group metal, and if present the carbon or graphite powder and binder, and drying at a temperature below that at which the enzyme is deactivated.

Claims

1. A method for the manufacture of enzyme electrodes comprising a substantially homogeneous mixture of a finely divided platinum group metal or oxide and one or more enzymes deposited as an electrically conductive layer upon the surface of an underlying support, the electrode being amperometrically responsive to the activity of the enzyme when the enzyme containing a layer of the electrode is in contact with the respective enzyme substrate, the method comprising forming a substantially uniform suspension comprising the finely divided platinum group metal or oxide and said enzyme(s) suspended in a liquid suspension medium, depositing that suspension on the surface of the support member, and drying the deposited suspension at a temperature below the deactivation temperature of the enzyme thereby to deposit the finely divided platinum group metal or oxide and said enzyme(s) as a substantially uniform, homogeneous coating layer on the surface of the support.

2. A method according to claim 1, wherein the suspension also contains a low temperature binder, said low temperature binder being effective to bind the finely divided platinum group metal or oxide and enzyme together to form a cohesive layer on the surface of the support at a temperature not exceeding the deactivation temperature of the enzyme(s), said layer comprising a substantially homogeneous dispersion of said finely divided platinum group metal or oxide and said enzyme.

3. A method according to claim 2, wherein the suspension also contains particles of finely divided carbon or graphite.

4. A method according to claim 3, wherein the particles of finely-divided platinum group metal or oxide are preadsorbed onto the surface of the finely divided carbon or graphite particles.

5. A method according to claim 4, wherein finely divided carbon or graphite particles have a particle size in the range 3 to 150 nm, and wherein the finely divided platinum group metal or oxide preadsorbed thereon has a particle size in the range 1 to 4 nm.

6. A method according to claim 1, wherein the liquid suspension medium is an aqueous medium.

7. A method according to claim 2, wherein the binder is gelatin or hydroxyethylcellulose.

8. A method according to claim 1, wherein the suspension is applied to the surface of the support member by a printing process.

9. A method according to claim 8, wherein the suspension is applied to the support member by screen printing.

10. A method according to claim 1, wherein the support member is itself electrically conductive.

11. A method according to claim 10, wherein the electrically conductive substrate is an electrically conductive carbon paper or carbon track.

12. A method according to claim 1, wherein the finely divided platinum group metal component is finely divided platinum or rhodium.

13. A method according to claim 1, wherein the enzyme used is glucose oxidase, lactate oxidase, or cholesterol oxidase.

14. In a method for the manufacture of enzyme electrodes comprising a substantially homogeneous mixture of a finely-divided platinum group metal or oxide adsorbed onto the surface of particles of a finely divided carbon or graphite powder and one or more enzymes deposited as a surface layer on the surface of an underlying electrically conductive support, the electrode being amperometrically responsive to the activity of the enzyme when the electrode is in contact with the respective enzyme substrate, the improvement which comprises forming an initial suspension comprising the enzyme, the finely divided carbon or graphite having said platinum group metal preadsorbed thereon, a binder operable to bind the enzyme and the platinum group metal or oxide containing carbon or graphite into a substantially uniform homogeneous layer on the surface of the support at a temperature below that at which the enzyme is deactivated, suspended in a liquid carrier, depositing the suspension as a uniform layer on the surface of the support, and drying the deposited layer at a temperature below the deactivation temperature of the binder.

15. A method according to claim 14, wherein the suspension is deposited on the surface of the support by printing.

16. A method according to claim 14, wherein the suspension is deposited on the surface of the support by screen printing.

17. A method according to claim 14, wherein the liquid carrier for the suspension is aqueous.

18. A method according to claim 17, wherein the binder is hydroxyethylcellulose or gelatin.

19. A method according to claim 14, wherein the electrically conductive support comprises an electrically conductive carbon paper.

20. A method according to claim 14, wherein the finely divided carbon or graphite comprises as said platinum group metal, finely divided platinum or rhodium.

21. A method according to claim 14, wherein the enzyme is glucose oxidase, lactate oxidase or cholesterol oxidase.

22. In a method for the manufacture of enzyme electrodes comprising a substantially homogeneous mixture of a finely-divided platinum group metal or oxide adsorbed onto the surface of particles of a finely divided carbon or graphite powder and one or more enzymes deposited as a surface layer on the surface of an underlying electrically conductive support, the electrode being amperometrically responsive to the activity of the enzyme when the electrode is in contact with the respective enzyme substrate, the improvement which comprises depositing as a surface layer on the surface of the electrically conductive support a suspension comprising the finely divided platinum group metal or oxide preadsorbed onto the surface of a finely divided graphite powder and a low temperature binder operable to bind the finely divided carbon or graphite powder and said enzyme into a uniform substantially homogeneous layer on the surface of the support at a temperature below the deactivation temperature of the enzyme, suspended in a liquid carrier, drying the deposited layer of platinum group metal or oxide containing carbon or graphite particles and said binder, impregnating the deposited layer with a suspension of the enzyme in a liquid carrier, and redrying the impregnated enzyme containing surface layer at a temperature below the deactivation temperature of the enzyme.

23. A method according to claim 22, wherein the carrier liquids for the platinised graphite or carbon suspension and the enzyme are both aqueous.

24. A method according to claim 23, wherein the binder is hydroxyethylcellulose or gelatin.

25. A method according to claim 22, wherein the support is an electrically conductive carbon paper.

26. A method according to claim 22, wherein the platinised carbon or graphite suspension is deposited on the support by screen printing.

27. An enzyme electrode comprising a substantially homogeneous mixture of a finely divided platinum group metal or oxide and one or more enzymes deposited as an electrically conductive surface layer on the surface of an underlying support member, said layer also containing a binder for the particles of finely divided platinum group metal or oxide and said enzyme(s), wherein said binder comprises a material effective to bind the finely divided platinum group metal or oxide particles and said enzyme(s) into a coherent, cohesive layer bonded to the support at a temperature not exceeding the deactivation temperature of the enzyme(s).

28. An enzyme electrode according to claim 27, wherein the binder is water-soluble or water-dispersible.

29. An enzyme electrode according to claim 28, wherein the binder is gelatin or hydroxyethylcellulose.

30. An enzyme electrode according to claim 27, wherein the enzyme containing layer also contains particles of finely divided carbon or graphite.

31. An enzyme electrode according to claim 30, wherein said finely divided particles of platinum group metal or oxide are preadsorbed onto the surface of the carbon or graphite particles.

32. An enzyme electrode according to claim 31, wherein the carbon or graphite particles have a particle size in the range 3 to 150 nm and the preadsorbed platinum group metal or oxide particles have a size in the range 1 to 4 nm.

33. An enzyme electrode according to claim 27, wherein the enzyme containing layer is printed onto the surface of the underlying support member.

34. An enzyme electrode according to claim 30, wherein the enzyme containing layer is screen printed onto the surface of the support member.

35. An enzyme electrode according to claim 27, wherein the underlying support member is itself electrically conductive.

36. An enzyme electrode according to claim 35, wherein the support member comprises a sheet of electrically conductive carbon paper or carbon track.

37. An enzyme electrode according to claim 27, wherein the enzyme containing layer contains as the platinum group metal component finely divided platinum or rhodium.

38. An enzyme electrode according to claim 27, wherein the enzyme containing layer contains a said enzyme glucose oxidase, lactate oxidase or cholesterol oxidase.

39. An enzyme electrode comprising a substantially homogeneous mixture containing the enzyme and a finely divided platinum group metal or oxide preadsorbed onto the surface of a finely divided carbon or graphite deposited on the surface of an electrically conductive support and containing a low temperature binder effective to bind the enzyme and finely divided platinum group metal or oxide containing particles into a uniform cohesive layer on the surface of the support at a temperature below the deactivation temperature of the enzyme.

40. An enzyme electrode according to claim 39, wherein the binder is water-soluble or water dispersible.

41. An enzyme electrode according to claim 40, wherein the binder is gelatin or hydroxyethylcellulose.

42. An enzyme electrode according to claim 39, wherein the electrically conductive support comprises an electrically conductive carbon paper or carbon track.

43. An enzyme electrode according to claim 39, wherein the layer of enzyme, platinised graphite or carbon and binder, is screen printed on the surface of the support.

44. An enzyme electrode according to claim 39, wherein the finely divided platinum group metal preadsorbed onto the surface of the finely divided graphite or carbon is platinum or rhodium.

45. An enzyme electrode according to claim 39, wherein the enzyme is glucose oxidase, lactate oxidase or cholesterol oxidase.