# US 8,569,004 B2 · Method for improving heat stability of composition containing water-soluble coenzyme-bound glucose dehydrogenase (GDH)

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- Number: US 8,569,004 B2
- Document ID: US8569004B2
- Status: Granted
- Kind: B2
- Application number: 12/573,516
- Filed: 2009-10-05
- Published: 2010-02-04
- Granted: 2013-10-29
- Earliest claimed: 2005-11-10
- Assignees: Toyo Boseki Kabushiki Kaisha
- Applicants: Masao Kitabayashi
- Inventors: Masao Kitabayashi
- Web page: https://patentgrep.com/patent/US8569004B2

## CPC

- A61B 5/14532: {for measuring glucose, e.g. by tissue impedance measurement}
- A61B 5/1486: using enzyme electrodes, e.g. with immobilised oxidase
- C12N 9/0006: {acting on CH-OH groups as donors (1.1)}
- C12N 9/96: Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
- C12Q 1/006: {for glucose}
- C12Q 1/32: involving dehydrogenase
- C12Q 1/54: involving glucose or galactose
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The invention relates to a method for improving the heat stability of glucose dehydrogenase (GDH). The invention provides a method for preparing a glucose sensor with soluble flavin-bound GDH by providing a composition comprising soluble flavin-bound GDH at an acidic pH and heat drying the composition onto a glucose sensor.

## Claims

### Claim 1

A method for preparing a glucose sensor with soluble flavin-bound glucose dehydrogenase, which method comprises (a) providing a composition comprising soluble flavin-bound glucose dehydrogenase at a pH in the range of 3.14 to 6.97 and (b) drying the composition onto a surface of a glucose sensor, wherein the soluble flavin-bound glucose dehydrogenase is from an Aspergillus microorganism.

### Claim 2 (depends on 1)

The method of claim 1, wherein the composition contains one or more dicarboxylic acids or salt compounds.

### Claim 3 (depends on 1)

The method of claim 1, wherein the composition contains one or more of compounds selected from the group consisting of sodium chloride, sodium sulfate, trisodium citrate, ammonium sulfate, succinic acid, malonic acid, glutaric acid, phthalic acid, and maleic acid.

### Claim 4

A method for preparing a glucose sensor with soluble flavin-bound glucose dehydrogenase, which method comprises (a) providing a composition comprising soluble flavin-bound glucose dehydrogenase at a pH in the range of 4.20 to 6.60 and (b) drying the composition onto a surface of a glucose sensor, wherein the soluble flavin-bound glucose dehydrogenase is from an Aspergillus microorganism.

### Claim 5 (depends on 1)

A glucose sensor prepared by the method of claim 1.

### Claim 6 (depends on 5)

A glucose sensor prepared by the method of claim 5.