Electrochemical system and method thereof

• US 9,605,295 B2
• Filed: 06/26/2012
• Issued: 03/28/2017
• Est. Priority Date: 11/07/2007
• Status: Active Grant

• Current Assignee: Cleveland State University
• Original Assignee: Cleveland State University
• Inventors: Yau, Siu-Tung
• Primary Examiner(s): Van, Luan
• Assistant Examiner(s): Kessel, Maris R
• Application Number:
  US13/533,248
• Publication Number:
  US 20120267255A1
• Time in Patent Office: 1,736 Days
• Field of Search: 20440301-40315, 205/775, 205/777.5, 205/792, 435/7.1, 435/287.2, 435/287.9
• US Class Current: 1/1
• 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 1/26 involving oxidoreductase
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48

Abstract

An embodiment of the invention provides an ultrasensitive and selective system and method for detecting reactants of the chemical/biochemical reaction catalyzed by an oxidoreductase, such as glucose and ethanol, at a concentration level down to zepto molar (10⁻²¹ M). In embodiments, the invention provides an ampyometric immuno-sensing system comprising a working electrode, an oxidoreductase, and an external voltage generator, wherein the oxidoreductase is immobilized on the working electrode; and the voltage generator generates a voltage to induce an electric field that permeates at least a portion of the interface between the oxidoreductase and the working electrode. The ultrasensitivity of the system and method is believed to be caused by the electrical field, which enhances the quantum mechanical tunneling effect in the interface, and therefore facilitates the interfacial electron transfer between the oxidoreductase and the working electrode.

Claims

1. An amperometric immuno-sensing system embodied in the form of an electrochemical cell system, the system comprising a working electrode, a reference electrode, and a counter electrode connected as a three-electrode cell, an enzyme-labeled antigen or an enzyme-labeled antibody, and an additional electrode, wherein the additional electrode is operatively connected to an external voltage source, wherein the enzyme-labeled antigen or the enzyme-labeled antibody is immobilized on the working electrode;

and wherein the additional electrode is configured to apply an external voltage from the external voltage source between the additional electrode and the working electrode to induce an electric field, at least a portion of an interface between the enzyme-labeled antigen or the enzyme-labeled antibody and the working electrode being permeated by the electric field.

2. The amperometric immuno-sensing system according to claim 1, further including a gating electrode.

3. The system of claim 2, wherein the gating electrodes are coated with insulators.

4. The amperometric immuno-sensing system according to claim 1, in which the external voltage ranges from about 0.001 volt to about 5 volt.

5. The amperometric immuno-sensing system according to claim 4, in which the external voltage ranges from about 0.005 volt to about 0.5volt.

6. The amperometric immuno-sensing system according to claim 5, in which the external voltage ranges from about 0.01 volt to about 0.2 volt.

7. The amperometric immuno-sensing system according to claim 4, wherein the system relies on the enzymatic activity of the enzyme.

8. The amperometric immuno-sensing system according to claim 1, wherein the working electrode is a modified graphite electrode.

9. The system of claim 1, wherein the system does not comprise mediators.

10. A voltammetry/amperometry system comprising a working electrode, a reference electrode, and a counter electrode connected as a three-electrode cell, an oxidoreductase, and an additional electrode, wherein the additional electrode is operatively connected to an external voltage source, wherein the oxidoreductase is immobilized on the working electrode;

and wherein the additional electrode is configured to apply a voltage from the external voltage source between the additional electrode and the working electrode to induce an electric field, at least a portion of an interface between the oxidoreductase and the working electrode being permeated by the electric field;

wherein the voltage source is configured to generate a V_ext from about 0.001 volt to about 2 volt.

11. The voltammetry/amperometry system according to claim 10, in which the voltage source is configured to generate a V_ext from about 0.005 volt to about 0.5 volt.

12. The voltammetry/amperometry system according to claim 11, in which the voltage source is configured to generate a Vext from about 0.01 volt to about 0.2 volt.

13. The voltammetry/amperometry system according to claim 10, in which the oxidoreductase comprises an enzyme that catalyzes the oxidation or reduction of an analyte or reactant resulting in charge transfer across the interface between the oxidoreductase and the working electrode.

14. The voltammetry/amperometry system according to claim 10, in which the oxidoreductase comprises a biological, organic or inorganic electro-active material that induces charge transfer at the interface between the electrode and the electro-active material.

15. The voltammetry/amperometry system according to claim 10, in which the oxidoreductase comprises an enzyme that catalyzes the transfer of electrons from a reductant to an oxidant.

16. The voltammetry/amperometry system according to claim 15, in which the reductant comprises CH—

OH group;

aldehyde or oxo;

CH—

CH group;

CH—

NH2group;

CH—

NH group;

NADH or NADPH;

sulfur group;

heme group;

diphenols;

hydrogen;

CH or CH2groups;

metal ions;

iron-sulfur proteins;

reduced flavodoxin;

phosphorus or arsenic;

or X—

H and Y—

H.

17. The voltammetry/amperometry system according to claim 15, in which the oxidant comprises peroxide or superoxide radical.

18. The voltammetry/amperometry system according to claim 10, in which the oxidoreductase comprises glucose oxidase enzyme (GOx).

19. The voltammetry/amperometry system according to claim 10, in which the oxidoreductase comprises alcohol dehydrogenase.

20. The voltammetry/amperometry system according to claim 10, wherein the electric field has a field intensity of up to about 100 volt/cm for inorganic material.

21. The voltammetry/amperometry system according to claim 10, wherein the electric field has a field intensity of up to 10 volt/cm for biochemical material such as oxidoreductase.

22. The voltammetry/amperometry system according to claim 21, in which the field intensity is from about 0.2 volt/cm to about 6.0 volt/cm.

23. The voltammetry/amperometry system according to claim 22, in which the field intensity is from about 0.4 volt/cm to about 3.0 volt/cm.

24. The voltammetry/amperometry system according to claim 10, which has a detection limit in the magnitude of zepto-molar (10′

21 M) range with zepto molar detection resolution.

25. A biosensor comprising the voltammetry/amperometry system according to claim 10.

26. An electrochemical sensor comprising the voltammetry/amperometry system according to claim 10.

27. The system of claim 10, wherein the system does not comprise mediators.