Potentiometric biosensor for detection of creatinine and forming method thereof

• US 20090194416A1
• Filed: 01/31/2008
• Published: 08/06/2009
• Est. Priority Date: 01/31/2008
• Status: Abandoned Application

• Current Assignee: Chung-Yuan Christian University
• Original Assignee: Chung-Yuan Christian University
• Inventors: Hsiung, Shen-Kan, Sun, Tai-Ping, Chou, Nien-Hsuan, Chou, Jung-Chuan
• Application Number:
  US12/024,066
• Publication Number:
  US 20090194416A1
• Time in Patent Office: Days
• US Class Current: 204/403.14
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: G01N 2333/986 acting on amide bonds in cyclic amides (3.5.2), e.g. beta-lactamase (penicillinase, 3.5.2.6), creatinine amidohydrolase (creatininase, EC 3.5.2.10), N-methylhydantoinase (3.5.2.6)
• CPC Class Codes: G01N 33/70 involving creatine or creatinine

Abstract

The present invention discloses a potentiometric biosensor for detecting creatinine, and the forming method thereof. The disclosed biosensor comprises a substrate, a working electrode formed on the substrate, a first reference electrode formed on the substrate, a second reference electrode formed on the substrate, and a packaging structure which separates the above-mentioned three electrodes. The working electrode comprises creatinine iminohydrolase (CIH). The detection signal is transmitted out from the biosensor for further processing through a wire or an exposed surface. The disclosed biosensor is replaceable.

Claims

1. A potentiometric biosensor for detection of creatinine, comprising:

a substrate;

a working electrode formed on said substrate;

a first reference electrode formed on said substrate; and

a second reference electrode formed on said substrate.

a packaging structure, which separates the above-mentioned three electrodes.

2. The potentiometric biosensor for detection of creatinine according to claim 1, wherein said substrate comprises one selected from the group consisting of the following:

insulating glass, non-insulated indium-tin oxide glass, non-insulated tin oxide glass and polyethylene terephthalate (PET).

3. The potentiometric biosensor for detection of creatinine according to claim 1, wherein said working electrode, comprising:

a first sensing layer formed on said substrate;

a first ion-selective layer formed on said first sensing layer; and

a first enzyme layer formed on said first ion-selective layer.

4. The potentiometric biosensor for detection of creatinine according to claim 3, wherein said first sensing layer is a non-insulated solid ion, comprising one selected from the group consisting of the following:

tin dioxide, titanium dioxide, and titanium nitride.

5. The potentiometric biosensor for detection of creatinine according to claim 3, wherein said first ion-selective layer is an ammonium ion-selective layer, comprising carboxylated polyvinylchloride (PVC-COOH).

6. The potentiometric biosensor for detection of creatinine according to claim 3, wherein said first enzyme layer comprises creatinine iminohydrolase (CIH).

7. The potentiometric biosensor for detection of creatinine according to claim 3, wherein said working electrode further comprises a first conducting layer which lies between said substrate and said first sensing layer for outward transmission of a detection signal, and said first conducting layer possesses a low impedance as to enhance the transmission efficiency of said detection signal, and said first conducting layer comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

8. The potentiometric biosensor for detection of creatinine according to claim 7, wherein said working electrode further comprises a wire connected to said first conducting layer to facilitate the transmission of said detection signal, and said wire comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

9. The potentiometric biosensor for detection of creatinine according to claim 3, wherein said first enzyme layer is immobilized on said first ion-selective layer via entrapment method by photocrosslinkable polyvinyl alcohol containing stilbazolium group (PVA-SbQ).

10. The potentiometric biosensor for detection of creatinine according to claim 7, wherein said first conducting layer comprises an exposed surface to electrically couple with the external world and for outward transmission of said detection signal.

11. The potentiometric biosensor for detection of creatinine according to claim 1, wherein said first reference electrode is an ammonium ion-selective electrode, comprising:

a second conducting layer formed on said substrate;

a second sensing layer formed on said second conducting layer; and

a second ion-selective layer formed on said second sensing layer.

12. The potentiometric biosensor for detection of creatinine according to claim 11, wherein said second conducting layer comprises an exposed surface to electrically couple with the external world and for outward transmission of a detection signal, and said second conducting layer possesses a low impedance as to enhance the transmission efficiency of said detection signal, and said second conducting layer comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

13. The potentiometric biosensor for detection of creatinine according to claim 11, wherein said first reference electrode further comprises a wire connected to said second conducting layer to facilitate the transmission of the detection signal, and said wire comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

14. The potentiometric biosensor for detection of creatinine according to claim 11, wherein said second sensing layer is a non-insulated solid ion, comprising one selected from the group consisting of the following:

tin dioxide, titanium dioxide, and titanium nitride.

15. The potentiometric biosensor for detection of creatinine according to claim 11, wherein said second ion-selective layer is an ammonium ion-selective layer, comprising carboxylated polyvinylchloride (PVC-COOH).

16. The potentiometric biosensor for detection of creatinine according to claim 1, wherein said second reference electrode is a hydrogen ion-selective electrode, comprising:

a third conducting layer formed on said substrate; and

a third sensing layer formed on said third conducting layer.

17. The potentiometric biosensor for detection of creatinine according to claim 16, wherein said third conducting layer comprises an exposed surface to electrically couple with the external world and for outward transmission of a detection signal, and said third conducting layer possesses a low impedance as to enhance the transmission efficiency of said detection signal, and said third conducting layer comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

18. The potentiometric biosensor for detection of creatinine according to claim 16, wherein said second reference electrode further comprises a wire connected to said third conducting layer to facilitate the transmission of said detection signal, and said wire comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

19. The potentiometric biosensor for detection of creatinine according to claim 16, wherein said third sensing layer is a non-insulated solid ion, comprising one selected from the group consisting of the following:

tin dioxide, titanium dioxide, and titanium nitride.

20. A working electrode for detection of creatinine, comprising:

a substrate;

a sensing layer formed on said substrate;

an ion-selective layer formed on said sensing layer; and

a enzyme layer formed on said ion-selective layer.

21. The working electrode for detection of creatinine according to claim 20, wherein said sensing layer is a non-insulated solid ion, comprising one selected from the group consisting of the following:

tin dioxide, titanium dioxide, and titanium nitride.

22. The working electrode for detection of creatinine according to claim 20, wherein said ion-selective layer is an ammonium ion-selective layer, comprising carboxylated polyvinylchloride (PVC-COOH).

23. The working electrode for detection of creatinine according to claim 20, wherein said enzyme layer comprises creatinine iminohydrolase (CIH).

24. The working electrode for detection of creatinine according to claim 20, wherein said working electrode further comprises a conducting layer which lies between said substrate and said sensing layer for outward transmission of a detection signal, and said conducting layer possesses a low impedance as to enhance the transmission efficiency of said detection signal, and said conducting layer comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

25. The working electrode for detection of creatinine according to claim 24, wherein said working electrode further comprises a wire connected to said conducting layer to facilitate the transmission of the detection signal, and said wire comprises one selected from the group consisting of the following:

copper, carbon, silver, aurum, silver chloride, Indium tin oxides (ITO).

26. The working electrode for detection of creatinine according to claim 20, wherein said enzyme layer is immobilized on said ion-selective layer via entrapment method by photocrosslinkable polyvinyl alcohol containing stilbazolium group (PVA-SbQ).

27. The working electrode for detection of creatinine according to claim 24, wherein said conducting layer comprises an exposed surface to electrically couple with the external world and for outward transmission of the detection signal.

28. A method for forming a working electrode to detect creatinine, comprising:

providing a substrate;

forming a conducting layer on said substrate;

forming a sensing layer on said conducting layer;

forming an ion-selective layer on said sensing layer; and

forming an enzyme layer on said ion-selective layer.

29. The method for forming a working electrode to detect creatinine according to claim 28, wherein said sensing layer is formed by deposition of tin oxide on said substrate through magnetron sputtering.