Method of measuring glucose concentration and glucose sensor with the use of glucose dehydrogenase

• US 7,341,846 B2
• Filed: 06/16/2003
• Issued: 03/11/2008
• Est. Priority Date: 06/17/2002
• Status: Expired due to Term

• Current Assignee: ARKRAY, Inc
• Original Assignee: ARKRAY, Inc
• Inventors: Yamaoka, Hideaki, Hoshijima, Mitsuhiro, Tsujimoto, Tomomichi
• Primary Examiner(s): Gitomer; Ralph
• Assistant Examiner(s): Shen; Bin
• Application Number:
  US10/518,858
• Publication Number:
  US 20060035300A1
• Time in Patent Office: 1,730 Days
• Field of Search: None
• US Class Current: 435/26
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: G01N 27/3272 Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels optical biosensors G01N33/52

Abstract

The present invention relates to a technique for measuring a glucose level by utilizing a reaction system containing an enzyme and an electron carrier. In accordance with the glucose level measuring method of the present invention, glucose dehydrogenase with cytochrome C attached thereto or glucose dehydrogenase derived from a microorganism belonging to a burkholderia genus is used as the enzyme, and a Ru compound is used as the electron carrier. The present invention further provides a glucose sensor in which glucose dehydrogenase with cytochrome C attached thereto or glucose dehydrogenase derived from derived from a microorganism belonging to a burkholderia genus is used as the enzyme, and a Ru compound is used as the electron carrier.

Claims

1. A glucose level measuring method using glucose dehydrogenase for measuring a glucose level by utilizing a reaction system containing glucose, an enzyme and an electron carrier,

wherein the enzyme is glucose dehydrogenase to which cytochrome C is attached and which is separate from the electron carrier;

and

wherein the electron carrier is a Ru compound represented by a chemical formula as follows;

Ru(NH₃)₅X]ⁿ⁺

where X represents NH₃, halogen ion, CN, pyridine, nicotinamide or H₂O, and n+ represents a valency of the Ru complex, which is determined by the kind of X, the method comprising;

allowing the cytochrome C attached glucose dehydrogenase to oxidize the glucose while reducing the Ru compound;

applying a voltage to the reaction system for the Ru compound to release electrons; and

detecting the electrons released by the Ru compound as a response current.

2. The glucose level measuring method according to claim 1, wherein the cytochrome C is derived from a microorganism belonging to a burkholderia genus.

3. The glucose level measuring method according to claim 1, wherein the cytochrome C has a molecular weight of about 43 kDa in SDS-polyacrylamide gel electrophoresis under a reduced condition.

4. The glucose level measuring method according to claim 1, wherein the glucose dehydrogenase includes an a subunit having a glucose dehydrogenase activity and a molecular weight of about 60 kDa in SDS-polyacrylamide gel electrophoresis under a reduced condition.

5. The glucose level measuring method according to claim 1, wherein the glucose dehydrogenase includes a γ

subunit having a molecular weight of about 14 kDa in SDS-polyacrylamide gel electrophoresis under a reduced condition.