Glucose sensor

• US 6,773,564 B1
• Filed: 09/28/1999
• Issued: 08/10/2004
• Est. Priority Date: 09/29/1998
• Status: Expired due to Term

• Current Assignee: PHC Holdings Corporation
• Original Assignee: Matsushita Electric Industrial Company Limited (Panasonic Holdings Corporation), Toyobo Company Limited
• Inventors: Iwata, Junko, Miyazaki, Shoji, Yoshioka, Toshihiko, Nankai, Shiro, Yugawa, Keiko, Baba, Hideyuki, Takeshima, Seiji
• Primary Examiner(s): Noguerola, Alex
• Application Number:
  US09/406,832
• Time in Patent Office: 1,778 Days
• Field of Search: 204/403, 204/403.01, 204/403.04, 204/403.09, 204/403.1, 204/403.11, 204/403.12, 204/403.14, 435/14, 435/189, 435/190, 205/777.5
• US Class Current: 204/403.14
• CPC Class Codes: C12Q 1/006 for glucose

Abstract

The present invention provides a high performance glucose sensor which can demonstrate high stability against preservation and produces only a low blank value. The glucose sensor comprises an electrically insulating base plate, an electrode system formed on the base plate, and a reaction layer which is formed in contact with or in the vicinity of the electrode system and contains at least a glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, the reaction layer further containing an additive such as phthalic acid.

Claims

1. A glucose sensor comprising an electrically insulating base plate, an electrode system including at least a working electrode and a counter electrode formed on said base plate, and a reaction layer which is formed in contact with or in the vicinity of said electrode system,

wherein said reaction layer contains phthalic acid or a phthalate, and a glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone.

2. The glucose sensor in accordance with claim 1, wherein said reaction layer further contains a stabilizer, said stabilizer having a function of retaining the activity of the enzyme and the long-term preservation of said sensor and decreasing the blank value of said sensor.

3. The glucose sensor in accordance with claim 2, wherein said stabilizer is a metal salt, an organic acid, a protein, an amide acid, a sugar or a derivative thereof, a surfactant, or ammonium sulfate.

4. The glucose sensor in accordance with claim 3, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, a strontium salt and a manganese salt.

5. The glucose sensor in accordance with claim 4, wherein said metal salt is a sulfate, a nitrate or a halide.

6. The glucose sensor in accordance with claim 4, wherein said calcium salt is CaCl₂.

7. The glucose sensor in accordance with claim 3, wherein said stabilizer is an organic acid selected from the group consisting of α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

8. The glucose sensor in accordance with claim 3, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

9. The glucose sensor in accordance with claim 3, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide and a derivative thereof, an oligosaccharide and a derivative thereof and a polysaccharide and a derivative thereof.

10. The glucose sensor in accordance with claim 9, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol, glucosamine and deoxyglucose, or a derivative thereof.

11. The glucose sensor in accordance with claim 9, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

12. The glucose sensor in accordance with claim 9, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

13. The glucose sensor in accordance with claim 9, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.

14. The glucose sensor in accordance with claim 3, said stabilizer is an amido acid selected from the group consisting of lysine, histidine, glutamic acid, glycylglycine and polylysine.

15. The glucose sensor in accordance with claim 3, wherein said stabilizer is a non-ionic surfactant.

16. The glucose sensor in accordance with claim 1, wherein said reaction layer further contains maleic acid, a maleate, succinic acid, a succinate, triethanol amine, a triethanol amine salt, citric acid, a citrate, dimethyl glutaric acid, 2-(N-morpholino) ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris (hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole or collidine.

17. A method for stabilizing glucose dehydrogenase for use in glucose sensors, wherein at least one additive is added to glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, said additive being selected from the group consisting of phthalic acid, a phthalate, triethanol amine, a triethanol amine salt, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

18. A method for stabilizing glucose dehydrogenase for use in glucose sensors, said glucose sensors each having a reaction layer comprising a glucose-dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, wherein phthalic acid or a phthalate is added to said glucose-dehydrogenase whose coenzyme is pyrrolo-quinoline quinone.

19. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 18, wherein a stabilizer is added to said glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, said stabilizer having a function of retaining the activity of enzyme and the long-term preservation of said sensor and decreasing the blank value of said sensor.

20. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 19, wherein said stabilizer is a metal salt, an organic acid, a protein, an amino acid, a sugar or a derivative thereof, a surfactant, or ammonium sulfate.

21. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, a strontium salt and a manganese salt.

22. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 21, wherein said metal salt is a sulfate, a nitrate or a halide.

23. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 21, wherein said calcium salt is CaCl₂.

24. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is an organic acid selected from the group consisting of α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

25. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

26. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide and a derivative thereof, an oligosaccharide and a derivative thereof, and a polysaccharide and a derivative thereof.

27. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 26, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol, glucosamine and deoxyglucose, or a derivative thereof.

28. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 26, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

29. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 26, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

30. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 26, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.

31. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is an amino acid selected from the group consisting of lysine, histidine, glutamic acid, glycylglycine and polylysine.

32. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 20, wherein said stabilizer is a non-ionic surfactant.

33. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with any of claim 18 to 32, wherein said reaction layer further contains maleic acid, a maleate, succinic acid, a succinate, triethanol amine, a triethanol amine salt, citric acid, a citrate, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole or collidine.

34. A glucose dehydrogenase composition for use in glucose sensors, said composition containing glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, and at lease one additive selected from the group consisting of phthalic acid, a phthalate, triethanol amine, a triethanol amine salt, dimethyl glutaric acid, (N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

35. A glucose dehydrogenase composition for use in glucose sensors, said composition containing glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone, and phthalic acid or a phthalate.

36. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 35, said composition further containing a stabilizer, said stabilizer having a function of retaining the activity of the enzyme and the long-term preservation of said sensor and decreasing the blank value of said sensor.

37. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 36, wherein said stabilizer is a metal salt, an organic acid, a protein, an amino acid, a sugar or a derivative thereof, a surfactant, or ammonium sulfate.

38. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, a strontium salt and a manganese salt.

39. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 38, wherein said metal salt is a sulfate, a nitrate or a halide.

40. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 38, wherein said calcium salt is CaCl₂.

41. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, wherein said stabilizer is an organic acid selected from the group consisting of a α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

42. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

43. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide and a derivative thereof, an oligosaccharide and a derivative thereof, and a polysaccharide and a derivative thereof.

44. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 43, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol glucosamine and deoxyglucose, or a derivative thereof.

45. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 43, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

46. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 43, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

47. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 43, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.

48. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, said stabilizer is an amino acid selected from the group consisting of lysine, histidine, glutamic acid, glycylglycine and polylysine.

49. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 37, wherein said stabilizer is a non-ionic surfactant.

50. The glucose dehydrogenase composition for use in glucose sensors in accordance with any of claims 35 to 49, wherein said composition further contains maleic acid, a maleate, succinic acid, a succinate, triethanol amine, a triethanol amine salt, citric acid, a citrate, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole or collidine.

51. A glucose sensor comprising an electrically insulating base plate, an electrode system including at least a working electrode and a counter electrode formed on said base plate, and a reaction layer which is formed in contact with or in the vicinity of said electrode system wherein said reaction layer contains:

a glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone; and

an additive selected from the group consisting of triethanol amine, a triethanol amine salt, citric acid, a citrate, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

52. The glucose sensor in accordance with claim 51, further comprising a at least one stabilizer selected from the group consisting of a metal salt, an organic acid, a protein, and a sugar and a derivative thereof.

53. The glucose sensor in accordance with claim 51, wherein the response of the sensor to glucose within the concentration range of 0 to 500 mg/dl is substantially linear after long term preservation.

54. A method for stabilizing glucose dehydrogenase for use in glucose sensors, wherein a stabilizer and a buffer are added to glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone,

said stabilizer being selected from the group consisting of a metal salt, an organic acid, a protein, and a sugar and a derivative thereof, and said buffer being selected from the group consisting of maleic acid, a maleate, triethanol amine, a triethanol amine salt, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

55. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 54, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, CaCl₂, a strontium salt and a manganese salt.

56. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 54, wherein said stabilizer is an organic acid selected from the group consisting of α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

57. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 54, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

58. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 54, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide and a derivative thereof, an oligosaccharide and a derivative thereof, and a polysaccharide and a derivative thereof.

59. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 58, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol, glucosamine and deoxyglucose, or a derivative thereof.

60. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 58, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

61. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 58, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

62. The method for stabilizing glucose dehydrogenase for use in glucose sensors in accordance with claim 58, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.

63. A glucose dehydrogenase composition for use in glucose sensors, said composition containing:

at least one stabilizer selected from the group consisting of a metal salt, an organic acid, a protein, and a sugar and a derivative thereof;

a glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone; and

a buffer selected from the group consisting of maleic acid, a maleate, triethanol amine, a triethanol amine salt, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

64. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 63, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, CaCl₂, a strontium salt and a manganese salt.

65. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 63, wherein said stabilizer is an organic acid selected from the group consisting of α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

66. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 63, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

67. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 63, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide and a derivative thereof, an oligosaccharide and a derivative thereof, and a polysaccharide and a derivative thereof.

68. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 67, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol, glucosamine and deoxyglucose, or a derivative thereof.

69. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 67, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

70. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 67, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

71. The glucose dehydrogenase composition for use in glucose sensors in accordance with claim 67, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.

72. A glucose sensor comprising an electrically insulating base plate, an electrode system including at least a working electrode and a counter electrode formed on said base plate, and a reaction layer which is formed in contact with or in the vicinity of said electrode system wherein said reaction layer contains:

at least one stabilizer selected from the group consisting of a metal salt, an organic acid, a protein, and a sugar and a derivative thereof;

a glucose dehydrogenase whose coenzyme is pyrrolo-quinoline quinone; and

an additive selected from the group consisting of maleic acid, a maleate, triethanol amine, a triethanol amine salt, citric acid, a citrate, dimethyl glutaric acid, 2-(N-morpholino)ethane sulfonic acid, a 2-(N-morpholino)ethane sulfonate, tris(hydroxymethyl)glycine, a tris(hydroxymethyl)glycine salt, tris(hydroxymethyl)aminomethane, a tris(hydroxymethyl)aminomethane salt, imidazole and collidine.

73. The glucose sensor in accordance with claim 72, wherein said stabilizer is a metal salt selected from the group consisting of a calcium salt, CaCl₂, a strontium salt and a manganese salt.

74. The glucose sensor in accordance with claim 72, wherein said stabilizer is an organic acid selected from the group consisting of α

-ketoglutaric acid, malic acid, fumaric acid, gluconic acid, cholic acid and deoxycholic acid.

75. The glucose sensor in accordance with claim 72, wherein said stabilizer is a protein selected from the group consisting of bovine serum albumin, egg albumin and gelatin.

76. The glucose sensor in accordance with claim 72, wherein said stabilizer is a sugar or a derivative thereof selected from the group consisting of a monosaccharide and a derivative thereof, a disaccharide, and a derivative thereof, an oligosaccharide and a derivative thereof, and a polysaccharide and a derivative thereof.

77. The glucose sensor in accordance with claim 72, wherein said stabilizer is a monosaccharide selected from the group consisting of glucose, fructose, galactose, mannose, xylose, inositol, monnitol, sorbitol, ribitol, glucosamine and deoxyglucose, or a derivative thereof.

78. The glucose sensor in accordance with claim 72, wherein said stabilizer is a disaccharide selected from the group consisting of sucrose, lactose, maltose and trehalose, or a derivative thereof.

79. The glucose sensor in accordance with claim 72, wherein said stabilizer is an oligosaccharide selected from the group consisting of malt triose, maltosyl cyclodextrin, α

-cyclodextrin, β

-cyclodextrin and γ

-cyclodextrin, or a derivative thereof.

80. The glucose sensor in accordance with claim 72, wherein said stabilizer is a polysaccharide selected from the group consisting of dextrin, amylose, glycogen, inulin and Ficoll, or a derivative thereof.