Multi-biosensor for GPT and got activity

• US 5,705,045 A
• Filed: 08/27/1996
• Issued: 01/06/1998
• Est. Priority Date: 08/29/1995
• Status: Expired due to Term

• Current Assignee: LG Electronics, Inc. (LG Corporation)
• Original Assignee: LG Electronics, Inc. (LG Corporation)
• Inventors: Lee, Kang Shin, Park, Je-Kyun
• Primary Examiner(s): Bell, Bruce F.
• Application Number:
  US08/703,537
• Time in Patent Office: 497 Days
• Field of Search: 204/403, 204/415, 435/817, 435/287.1, 435/289.1
• US Class Current: 204/403.1
• 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: C12Q 1/52 involving transaminase
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

Multi-biosensor in which only one enzyme is used for handy and simultaneous measurement of GPT and GOT activities, and a method for fabricating the multi-biosensor, are disclosed, the biosensor including an insulating substrate; a reference electrode formed on the insulating substrate; first and second working electrodes formed on one side of the insulating substrate having the reference electrode formed thereon; third and fourth working electrodes formed on the other side of the insulating substrate having the reference electrode formed thereon opposite to the first and second working electrodes centered on the reference electrode; first, second, third and fourth carbon paste layers formed on the first, second, third and fourth working electrodes respectively; first and second enzyme/polymer carbon paste layers formed on the first and second carbon paste layers respectively; first and second non-enzyme/polymer carbon paste layers formed on the third and fourth carbon paste layers respectively; an amino acid/polymer paste layer for making an enzymatic reaction with GPT formed extended to the first enzyme/polymer carbon paste layer and the first non-enzyme/polymer carbon paste layer; and, an amino acid/polymer paste layer for making an enzymatic reaction with GOT formed extended to the second enzyme/polymer carbon paste layer and the second non-enzyme/polymer carbon paste layer.

Claims

1. A biosensor comprising:

an insulating substrate;

a reference electrode formed on the insulating substrate;

first and second working electrodes formed on one side of the insulating substrate having the reference electrode formed thereon;

third and fourth working electrodes formed on the same plane of the insulating substrate but parallel to the first and second working electrodes centered on the reference electrode;

first, second, third and fourth carbon paste layers formed on the first, second, third and fourth working electrodes respectively;

first and second enzyme/polymer carbon paste layers formed on the first and second carbon paste layers respectively;

first and second non-enzyme/polymer carbon paste layers formed on the third and fourth carbon paste layers respectively;

an amino acid/polymer paste layer for making an enzymatic reaction with Glutamate Pyruvate Transaminase (GPT) formed covering the first enzyme/polymer carbon paste layer and the first non-enzyme/polymer carbon paste layer; and

,

an amino acid/polymer paste layer for making an enzymatic reaction with Glutamate Oxaloacetate Transaminase (GOT) formed covering the second enzyme/polymer carbon paste layer and the second non-enzyme/polymer carbon paste layer.

2. A biosensor as claimed in claim 1, wherein the insulating substrate is formed of a polymer selected from the group consisting of polyester, Polyvinyl Chloride (PVC) and polycarbonate.

3. A biosensor as claimed in claim 1, wherein each of the first and second enzyme/polymer carbon paste layers includes glutamate oxidase, hydroxyethyl cellulose and carbon powder.

4. A biosensor as claimed in claim 1, wherein each of the first and second non-enzyme/polymer carbon paste layers includes bovine serum albumin, hydroxyethyl cellulose and carbon powder.

5. A biosensor as claimed in claim 1, wherein each of the enzyme/polymer carbon paste layers includes glutamate dehydrogenase, NAD⁺, hydroxyethyl cellulose and carbon powder.

6. A biosensor as claimed in claim 1, wherein each of the non-enzyme/polymer carbon paste layers includes bovine serum albumin, NAD⁺, hydroxyethyl cellulose and carbon powder.

7. A biosensor as claimed in claim 1, further comprising a guide layer for forming grooves for separation and introduction of a sample, the guide layer formed for adhering both to the electrode part and an outer protection plate, the outer protection plate formed of an insulating substrate.

8. A biosensor as claimed in claim 7, wherein the electrode part includes two working electrodes for measuring GPT and two working electrodes for measuring GOT and a reference electrode.

9. A biosensor as claimed in claim 8, wherein the working electrodes for measuring GPT and the working electrodes for measuring GOT are of differential amplification types.

10. A method for fabricating a biosensor comprising the steps of:

providing an insulating substrate;

forming a reference electrode on the insulating substrate;

forming first and second working electrodes on one side of the insulating substrate having the reference electrode formed thereon;

forming third and fourth working electrodes on the same plane of the insulating film but parallel to the first and second working electrodes centered on the reference electrode;

forming first, second, third and fourth carbon paste layers on the first, second, third and fourth working electrodes, respectively;

forming first and second enzyme/polymer carbon paste layers on the first and second carbon paste layers, respectively;

forming first and second non-enzyme/polymer carbon paste layers on the third and fourth carbon paste layers respectively;

forming an amino acid/polymer paste layer for enzymatic reaction with Glutamate Pyruvate Transaminase (GPT) covering the first enzyme/polymer carbon paste layer and the first non-enzyme/polymer carbon paste layer; and

,

forming an amino acid/polymer paste layer for making an enzymatic reaction with Glutamate Oxaloacetate Transaminase (GOT) covering the second enzyme/polymer carbon paste layer and the second non-enzyme/polymer carbon paste layer.

11. A method as claimed in claim 10, wherein the insulating substrate is formed of a polymer selected from the group consisting of polyester, Polyvinyl Chloride (PVC) and polycarbonate.

12. A method as claimed in claim 10, wherein each of the first and second enzyme/polymer carbon paste layers includes glutamate oxidase, hydroxyethyl cellulose and carbon powder.

13. A method as claimed in claim 10, wherein each of the first and second non-enzyme/polymer carbon paste layers includes bovine serum albumin, hydroxyethyl cellulose and carbon powder.

14. A method as claimed in claim 10, wherein each of the enzyme/polymer carbon paste layers includes glutamate dehydrogenase, NAD⁺, hydroxyethyl cellulose and carbon powder.

15. A method as claimed in claim 10, wherein each of the non-enzyme/polymer carbon paste layers includes bovine serum albumin, NAD⁺, hydroxyethyl cellulose and carbon powder.

16. A method as claimed in claim 10, wherein the amino acid/polymer paste layer for making an enzymatic reaction with GPT includes L-alanine, α

- ketoglutarate and hydroxyethyl cellulose.

17. A method as claimed in claim 10, wherein the amino acid/polymer paste layer for making an enzymatic reaction with GPT includes hydroxyethyl cellulose solution containing about 1M L-alanine and 100 mM α

- ketoglutarate.

18. A method as claimed in claim 10, wherein the amino acid/polymer paste layer for making an enzymatic reaction with GOT includes L-aspartate, α

- ketoglutarate and hydroxyethyl cellulose.

19. A method as claimed in claim 10, wherein the amino acid/polymer paste layer for making an enzymatic reaction with GOT includes hydroxyethyl cellulose solution containing about 500 mM L-aspartate and 100 mM α

- ketoglutarate.

20. A method as claimed in claim 10, further comprising the step of forming a guide layer for forming grooves for separation and introduction of a sample, the guide layer formed for adhering both to the electrode part and an outer protection plate, the outer protection plate formed of an insulating substrate.