Biosensor

• US 6,897,035 B1
• Filed: 07/06/2000
• Issued: 05/24/2005
• Est. Priority Date: 07/06/1999
• Status: Expired due to Fees

• Current Assignee: Forskarpatent i Syd AB (Government of Sweden)
• Original Assignee: Forskarpatent i Syd AB (Government of Sweden)
• Inventors: Csöregi, Elisabeth, Niculescu, Mihaela, Frebort, Ivo
• Primary Examiner(s): Witz, Jean C.
• Assistant Examiner(s): HANLEY, SUSAN MARIE
• Application Number:
  US10/019,651
• Time in Patent Office: 1,783 Days
• Field of Search: 435/25, 435/28, 435/283.1, 435/287.1, 435/287.9, 435/817, 435/7.9
• US Class Current: 435/25
• CPC Class Codes: C12Q 1/004 mediator-assisted
• 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: Y10S 435/817 Enzyme or microbe electrode

Abstract

The present invention relates to a biosensor for the detection and/or the determination of freshness biomarkers, such as biogenic amines (preferably histamine) in food and beverage, comprising an electrode and a mono-enzyme system, such as an amine oxidase, or a bi-enzyme system of an amine oxidase and a peroxidase. The enzymes are optionally crosslinked into an osmium based redox polymer.

Claims

1. A method for the detection or determination of histamine in a sample of microdialysates or dialysates, comprising the steps of:

(A) applying said sample to a biosensor comprising an electrode and (i) a mono-enzyme system of an amine oxidase, which is a copper-containing grass pea oxidase (E. C. 1.4.3.6) or (ii) a bi-enzyme system of an amine oxidase, which is a copper-containing grass pea oxidase (E.C. 1.4.3.6), coupled with horseradish, soybean, tobacco, sweet potato or palmtree peroxidase;

wherein said electrode is a carbon/graphite based electrode, and whereby said amine oxidase is cross-linked to the electrode into an osmium based redox polymer; and

(B) detecting an electrical output from said biosensor.

2. The method according to claim 1, wherein the peroxidase is horseradish peroxidase.

3. The method according to claim 1, wherein the osmium based redox polymer comprises poly(1-vinylimidazole) complexed with [Os(4,4′

-dimethyl-bi-pyridin)₂Cl]^+/2+ and polyethyleneglycol)diglycidyl ether as the cross-linking agent.

4. The method according to claim 1, wherein biosensor is of Type I, Type II or Type III type of biosensor, wherein:

Type I is the mono-enzyme or the bi-enzyme system is added directly into the electrode surface;

or

Type II is the mono-enzyme or the bi-enzyme system is entrapped in the osmium based redox polymer added on the surface of the electrode;

or

Type III is the mono-enzyme or the bi-enzyme system and the osmium based redox polymer forms sequential coatings added on the surface of the electrode.

5. The method according to claim 4, wherein the biosensor of Type III is one of Type III a, Type III b, Type III c or Type III d, wherein:

Type III a is a second coating of the mono-enzyme is coating a dried layer of peroxidase and redox hydrogel;

or

Type III b is a second coating of peroxidase and redox hydrogel is coating a dried layer of the mono-enzyme;

or

Type III c is a second coating of the mono-enzyme entrapped in redox hydrogel is coating a dried layer of peroxidase;

or

Type III d is a second coating of peroxidase is coating a dried layer of mono-enzyme entrapped in redox hydrogel.

6. The method according to claim 2, wherein the weight ratio of amine oxidase to horseradish peroxidase is 80:

20.