Reagents for electrochemical test strips

• US 10,760,111 B2
• Filed: 07/25/2012
• Issued: 09/01/2020
• Est. Priority Date: 07/27/2011
• Status: Active Grant

• Current Assignee: Agamatrix Incorporated (AgaMatrix Holdings LLC d/b/a WaveForm Diabetes)
• Original Assignee: Agamatrix Incorporated (AgaMatrix Holdings LLC d/b/a WaveForm Diabetes)
• Inventors: Harding, Ian, Lee, Mary Y., Tan, Sandie
• Primary Examiner(s): Kaur, Gurpreet
• Application Number:
  US13/557,372
• Publication Number:
  US 20130026050A1
• Time in Patent Office: 2,960 Days
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: G01N 33/521 Single-layer analytical elements

Abstract

A dry reagent composition that includes an active redox enzyme that oxidizes an analyte as a specific substrate to produce an inactive reduced form of the enzyme; and a salt of ferricyanide provides improved performance in electrochemical test strips such as those used for detection of glucose. The salt of ferricyanide consists of ferricyanide and positively-charged counter ions, and the positively charged counter ions are selected such that the salt of ferricyanide is soluble in water, and such that the salt of ferricyanide or the crystalline phase of the salt of ferricyanide has a solubility in water and/or a lower E⁰_eff at a concentration of 100 mM than potassium ferricyanide. For example, the salt of ferricyanide may be tetramethylammonium ferricyanide.

Claims

1. A dry reagent composition comprising:

(a) an active redox enzyme that oxidizes an analyte as a specific substrate to produce an inactive reduced form of the enzyme; and

(b) tris-tetramethylammonium ferricyanide.

2. The dry reagent of claim 1, wherein the dry reagent further comprises an electron transfer mediator in oxidized form that is not a salt of ferricyanide, said oxidized form of electron transfer mediator having an electrochemical potential in aqueous medium sufficient to oxidize the inactive reduced form of the enzyme to regenerate active redox enzyme.

3. The dry reagent of claim 2, wherein the oxidized form of the electron transfer mediator is an osmium coordination complex.

4. The dry reagent of claim 3, wherein the osmium coordination complex is Os(dmbpy)₂Im₂Cl.

5. The dry reagent of claim 3, wherein the osmium coordination complex is Os(dmbpy)₂PicCl.

6. The dry reagent of claim 5, further comprising a reduced form of the electron transfer mediator, wherein the amount of said reduced form of the electron transfer mediator relative to the oxidized form of the electron transfer mediator is such that a solution of the reagent has a baseline comparable to a steady-state baseline signal that is produced by a solution of a reagent in the absence of the reduced form of the electron transfer mediator that has been aged in a dessicator for 14 days at 50°

C.

7. The dry reagent of claim 5, further comprising tetra-tetramethylammonium ferrocyanide, wherein the amount of ferrocyanide relative to ferricyanide is such that a solution of dry reagent with the tetra-tetramethylammonium ferrocyanide has a baseline comparable to a steady-state baseline signal that is produced by a solution of a dry reagent containing just the ferricyanide and enzyme components of the dry reagent that has been aged in a dessicator for 14 days at 50°

C.

8. The dry reagent of claim 5, further comprising a zwitterion buffering agent comprising a positively-charged buffer counter ion and a buffer conjugate base.

9. The dry reagent of claim 1, further comprising a zwitterion buffering agent comprising a positively-charged buffer counter ion and a buffer conjugate base.

10. The dry reagent of claim 1, wherein the active redox enzyme is glucose oxidase.

11. The dry reagent of claim 1, wherein essentially all of cations in the dry reagent are tetramethylammonium cations.

12. The dry reagent of claim 11, wherein the active redox enzyme is glucose oxidase.

16. The dry reagent of any claim 8, further comprising a zwitterionic wetting agent comprising a hydrophilic head group including an amine and a sulphonate, and a hydrophobic aliphatic tail of 10 to 16 carbon atoms.

17. The dry reagent of claim 16, wherein the hydrophobic tail of the zwitterionic wetting agent is a 12 carbon atom tail.

18. The dry reagent of claim 17, wherein the active redox enzyme is glucose oxidase.

22. The dry reagent of claim 9, further comprising a zwitterionic wetting agent comprising a hydrophilic head group including an amine and a sulphonate, and a hydrophobic aliphatic tail of 10 to 16 carbon atoms.

23. The dry reagent of claim 22, wherein the hydrophobic tail of the zwitterionic wetting agent is a 12 carbon atom tail.

24. The dry reagent of claim 23, wherein the active redox enzyme is glucose oxidase.

25. A test strip comprising:

(a) first and second non-reactive electrodes;

(b) a sample cell for receiving a liquid sample, wherein a liquid sample disposed within the sample cell is in contact with the first and second electrodes; and

(c) a dry reagent in accordance with claim 1, wherein said dry reagent is disposed such that upon application of a liquid sample to the test strip the dry reagent dissolves in the sample within the sample cell.

26. The test strip of claim 25, wherein the first and second electrodes are metal electrodes.

27. The test strip of claim 25, wherein at least one of the first and second electrodes comprises palladium.

28. A method for testing for an analyte in a liquid sample comprising the steps of:

(a) applying a liquid sample to a test strip in accordance with claim 26;

wherein the enzyme in the dried reagent is selected to be specific for the analyte;

(b) applying an external signal to the test strip to generate a signal indicative of the amount of analyte in the sample.

29. The method of claim 28, wherein the external signal is a potential that is applied between the first and second electrodes of the test strip, and a current is measured as the signal indicative of the amount of analyte in the sample.

13. A dry reagent composition comprising:

(a) an active redox enzyme that oxidizes an analyte as a specific substrate to produce an inactive reduced form of the enzyme;

(b) tris-tetramethylammonium ferricyanide;

(c) an electron transfer mediator in oxidized form that is not a salt of ferricyanide, said oxidized form of electron transfer mediator having an electrochemical potential in aqueous medium sufficient to oxidize the inactive reduced form of the enzyme to regenerate active redox enzyme, wherein the electron transfer mediator is Os(dmbpy)₂PicCl, and

(d) a zwitterion buffering agent comprising a positively-charged buffer counter ion and a buffer conjugate base, wherein essentially all of the positively-charged buffer counter ion is tetramethylammonium.

14. The dry reagent of claim 13, wherein the buffer conjugate base is the conjugate base of 3[N-Tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid.

15. The dry reagent of claim 13, wherein the buffer conjugate base is the conjugate base of N-Tris(hydroxy-methyl)methyl-2-aminoethanesulfonic acid.

19. A dry reagent composition comprising:

an active redox enzyme that oxidizes an analyte as a specific substrate to produce an inactive reduced form of the enzyme;

(b) tris-tetramethylammonium ferricyanide; and

(c) a zwitterion buffering agent comprising a positively-charged buffer counter ion and a buffer conjugate base, wherein essentially all of the positively-charged buffer counter ion is tetramethylammonium.

20. The dry reagent of claim 19, wherein the buffer conjugate base is the conjugate base of 3-[N-Tris(hydroxymethyl)methylamino]-2-hydroxypropanesulfonic acid.

21. The dry reagent of claim 19, wherein the buffer conjugate base is the conjugate base of N-Tris(hydroxy-methyl)methyl-2-aminoethanesulfonic acid.

30. A method for forming a dry shelf stable electrochemical test reagent for use in detection of an analyte comprising the steps of:

(a) selecting an oxidoreductase enzyme that oxidizes/reduces the analyte as a specific substrate;

(b) combining the selected enzyme and tris-tetramethylammonium ferricyanide to form a dry shelf stable electrochemical test reagent for use in detection of the analyte, wherein the selected enzyme is glucose oxidase.