Enzyme electrode and method for producing the same

• US 8,172,996 B2
• Filed: 04/09/2009
• Issued: 05/08/2012
• Est. Priority Date: 02/27/2009
• Status: Expired due to Fees

• Current Assignee: Chung-Yuan Christian University
• Original Assignee: Chung-Yuan Christian University
• Inventors: Cheng, Chean-Yeh, Chang, Kuo-Chung
• Primary Examiner(s): Olsen, Kaj K
• Application Number:
  US12/421,565
• Publication Number:
  US 20100219072A1
• Time in Patent Office: 1,125 Days
• Field of Search: 204/403.01, 204/403.11, 204/403.14, 427/2.13
• US Class Current: 204/403.11
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C25D 3/48 of gold
• CPC Class Codes: C25D 7/00 Electroplating characterised by the article coated

Abstract

An enzyme electrode and its producing method are disclosed. The method includes providing a substrate having a carbon surface; forming a gold surface on the carbon surface and forming an electrode; modifying the gold surface by L-cysteine; modifying the gold surface by N,N′-dicyclohexylcarbodiimide; and chemically bonding the modified gold surface with a glucose oxidase.

Claims

1. A method for producing an enzyme electrode used to detect glucose, comprising:

providing a substrate having a carbon surface, wherein the substrate is a pencil lead, and the carbon surface is formed by coating a carbon paste on the surface of the pencil lead;

forming a gold surface on the carbon surface and thus forming an electrode;

modifying the gold surface by L-cysteine to bond covalently the gold particles of the gold surface and the sulphydryl group of L-cysteine, thereby forming a first electrode having a first modified surface;

modifying the first modified surface by N,N′

-dicyclohexylcarbodiimide, the L-cysteine of the first modified surface and the N,N′

-dicyclohexylcarbodiimide being dehydrated to form covalent bonds, thereby forming a second electrode having a second modified surface; and

contacting the second modified surface with the glucose oxidase, an amide bond being formed between the L-cysteine group of the second modified surface and the glucose oxidase, thereby forming a third electrode having a third modified surface.

2. The method as recited in claim 1, further comprising a thermal process at about 120°

C. for drying the surface of the carbon paste, after the carbon paste is coated on the surface of the pencil lead.

3. The method as recited in claim 1, wherein the gold surface is formed by a deposition, ink-inject, electrodeposition, or screen-printing method.

4. The method as recited in claim 1, wherein the gold surface is formed by electrodeposition in which the gold ions are reduced and deposited on the carbon surface.

5. An enzyme electrode used to detect glucose and produced by the method as follows:

providing a substrate having a carbon surface, wherein the substrate is a pencil lead, and the carbon surface is formed by coating a carbon paste on the surface of the pencil lead;

forming a gold surface on the carbon surface and thus forming an electrode;

modifying the gold surface by L-cysteine to bond covalently the gold particles of the gold surface and the sulphydryl group of L-cysteine, thereby forming a first electrode having a first modified surface;

modifying the first modified surface by N,N′

-dicyclohexylcarbodiimide, the L-cysteine of the first modified surface and the N,N′

-dicyclohexylcarbodiimide being dehydrated to form covalent bonds, thereby forming a second electrode having a second modified surface; and

contacting the second modified surface with the glucose oxidase, an amide bond being formed between the L-cysteine group of the second modified surface and the glucose oxidase, thereby forming the enzyme electrode.

6. The enzyme electrode as recited in claim 5, wherein the enzyme electrode is applied to an electrochemical sensor for the detecting of glucose, the electrochemical sensor comprising a working electrode and a reference electrode, and the enzyme electrode functioning as the working electrode.

7. The enzyme electrode as recited in claim 6, wherein the electrochemical sensor further comprises a counter electrode.

8. The enzyme electrode as recited in claim 6, further comprising a mediator coated on the surface of the working electrode.

9. The enzyme electrode as recited in claim 6, wherein the mediator comprises hydroxylmethylferrocene (Fc).

10. An enzyme electrode used to detect glucose, comprising:

a substrate structure, said substrate structure comprising a pencil lead, a carbon paste coated on the surface of the pencil lead, and a gold layer covering the carbon paste;

a modified structure, said modified structure being chemically bound with the gold layer, the modified structure comprising a L-cysteine group and a glucose oxidase group, an Au—

S covalent bond being formed between the gold layer and the L-cysteine group, and an amide bond being covalently bound between the L-cysteine group and the glucose oxidase group.

11. The enzyme electrode as recited in claim 10, wherein the enzyme electrode is applied to an electrochemical sensor for the detecting glucose, the electrochemical sensor comprises a working electrode and a reference electrode, and the enzyme electrode functions as the working electrode.

12. The enzyme electrode as recited in claim 11, wherein the electrochemical sensor further comprises a counter electrode.

13. The enzyme electrode as recited in claim 11, further comprising a mediator coated on the surface of the working electrode.

14. The enzyme electrode as recited in claim 11, wherein the mediator comprises hydroxylmethylferrocene (Fc).