Monitoring bioelectrochemical reactions and novel media for bioelectrochemical reactions

• US 5,521,101 A
• Filed: 08/26/1993
• Issued: 05/28/1996
• Est. Priority Date: 08/28/1992
• Status: Expired due to Fees

• Current Assignee: Cranfield University
• Original Assignee: Cranfield University
• Inventors: Saini, Selwayan, Turner, Anthony P. F.
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/112,670
• Time in Patent Office: 1,006 Days
• Field of Search: 204/403, 204/410, 204/411-412, 204/414-415, 204/418, 204/421, 204/431, 422/82.01-82.02, 422/90, 422/98, 427/2, 435/4, 435/7.1, 435/7.9, 435/14, 435/25, 435/26, 435/176, 435/177, 435/180, 435/288, 435/291, 436/518, 436/530, 436/531, 436/535, 436/152
• US Class Current: 205/777.5
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/002 Electrode membranes
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

A method of determining an analyte in the gaseous or vapour phase and in which a bioreceptor or biomimic is retained at an electrode. The bioreceptor or biomimic is preferably retained at a support at the electrode which comprises a solid or gel matrix of an electrolyte, especially organic salt electrolytes. Electrochemical detection of analytes in this way has several advantages over existing methods which rely on solution monitoring. For example gas sensors can be prepared for monitoring an analyte by the occurrence of a reaction with a bioreceptor or biomimic, in addition to monitoring the presence of toxins due to inhibition of the bioreceptor or biomimic reaction. Furthermore, the invention enables gas or vapour analyte monitoring with increased sensitivity and speed and greater stability of the sensors can be achieved. The invention also relates to novel media for carrying out bioelectrochemical reactions.

Claims

1. A method for determining a gaseous or vapor analyte comprising

(1) providing a substantially liquid-free electrochemical cell comprising microvoltammetric electrodes including a microvoltammetric sensing electrode;

solid or semi-solid support consisting essentially of an electrolyte in electrical contact with the microvoltammetric sensing electrode; and

an enzyme or synzyme;

whereto the enzyme or synzyme is immobilized within the support in electrical contact with the microvoltammetric sensing electrode and wherein the enzyme or synzyme is contactable with a gaseous or vapor phase, and

(2) exposing the electrochemical cell to a gaseous or vapor phase suspected of containing the gaseous or vapor analyte which is a substrate for the enzyme or synzyme, whereby the gaseous or vapor analyte contacts the enzyme or synzyme and reacts thereby generating a detectable electrical response relatable to presence of the analyte; and

(3) determining the electrical response by the microvoltammetric sensing electrode to determine the presence or amount of the analyte.

2. A method according to claim 1 wherein the detectable electrical response increases on exposing the electrochemical cell to an increasing concentration of the analyte.

3. A method according to claim 1 in which the enzyme or synzyme is an enzyme for a redox reaction.

4. A method according to claim 1 wherein the solid or semi-solid support has a thickness no greater than 1 mm.

5. A method according to claim 4 in which the electrolyte is an organic salt.

6. A method according to claim 5 in which the organic salt comprises a tetrabutyl ammonium salt or a tetra ethyl ammonium salt.

7. A method according to claim 5 in which the organic salt is chosen from tetrabutyl ammonium perchlorate, tetrabutyl ammonium toluene-4-sulphonate, tetrabutylammonium methane sulphonate, tetrabutylammonium phenol borate, tetraethylammonium tetrafluoroborate, tetrabutyl-ammonium chloride and tetrabutyl ammonium iodide.

8. A method according to claim 4 in which the enzyme or synzyme is an enzyme for a redox reaction.

9. A method according to claim 1 wherein the support is formed from a hygroscopic electrolyte.

10. A method for determining a gaseous or vapor analyte comprising

(1) providing a substantially liquid-free electrochemical cell comprising microvoltammetric electrodes including a microvoltammetric sensing electrode;

a solid or semi-solid support in electrical contact with the microvoltammetric sensing electrode; and

an enzyme or synzyme;

whereto the support consists essentially of a solid or gel matrix formed from an electrolyte, wherein the enzyme or synzyme is retained by the support as an underlayer or overlayer for the support, and the support is in electrical contact with the microvoltammetric sensing electrode and whereto the enzyme or synzyme is contactable with a gaseous or vapor phase, and

(2) exposing the electrochemical cell to a gaseous or vapor phase suspected of containing the gaseous or vapor analyte which is a substrate for the enzyme or synzyme, whereby the gaseous or vapor analyte contacts the enzyme or synzyme and reacts thereby generating a detectable electrical response relatable to presence of the analyte; and

(3) detecting the electrical response by the microvoltammetric sensing electrode to determine the presence or amount of the analyte.

11. A method according to claim 10 wherein the enzyme or synzyme is retained by the support as an traderlayer for the support.

12. A method according to claim 10, said matrix comprising a solid or gel matrix formed from a tetraalkyl ammonium salt selected from tetrabutyl ammonium salts and tetraethylammmonium salts.

13. A substantially liquid-free support for a bioelectrochemical reaction comprising a solid or gel matrix comprising a tetra butyl ammonium salt or a tetra ethyl ammonium salt and an enzyme or synzyme, said matrix being permeable to a gaseous reactant, the reactant being a substrate for the enzyme or synzyme, wherein the enzyme or synzyme is retained by the support and wherein the enzyme or synzyme is contactable by the gaseous reactant.

14. A microvoltammetric sensing electrode comprising a conductor, a solid or semi-solid support m electrical contact with the conductor and an enzyme or synzyme retained by the support, the support consisting essentially of a solid or gel matrix of all electrolyte, wherein the enzyme or synzyme is immobilized within the support and the support is permeable to a gaseous reactant which is a substrate for the enzyme or synzyme and whereto the electrolyte is an organic salt.

15. An electrochemical sensor comprising sensing and counter electrodes, means for permitting access of analyte to the sensing electrode, and wherein the sensing electrode is as defined in claim 14.

16. A sensor according to claim 15 in which the sensing electrode and counter electrode are provided by an interdigitated microelectrode.

17. A sensing electrode according to claim 14 in which the organic salt is chosen from tetrabutyl ammonium perchlorate, tetrabutyl ammonium toluene-4-sulphonate, tetrabutylammonium methane sulphonate, tetrabutyl ammonium phenol borate, tetraethylammonium tetrafluoroborate, tetrabutylammonium chloride and tetrabutyl ammonium iodide.