Disposable sub-microliter volume sensor and method of making

• US 6,258,229 B1
• Filed: 06/02/1999
• Issued: 07/10/2001
• Est. Priority Date: 06/02/1999
• Status: Expired due to Term

• Current Assignee: Nova Biomedical Corporation (Nova Chemicals)
• Original Assignee: Nova Biomedical Corporation (Nova Chemicals)
• Inventors: Young, Chung Chang, Cai, Xiaohua, Seto, Fung, Winarta, Handani
• Primary Examiner(s): Tung, T.
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/324,493
• Time in Patent Office: 769 Days
• Field of Search: 204/400, 204/403, 204/412, 204/416, 204/431, 204/280, 204/286, 422/82.01, 422/88.02, 422/82.03, 324/439, 427/2.13, 216/13
• US Class Current: 204/403.04
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose

Abstract

A disposable electrode strip for testing a fluid sample including a laminated strip with a first and second end, a vent, an open path for receiving a fluid sample of less than one microliter beginning from the first end and connecting to the vent, a working electrode, a reference electrode and a pseudo-working electrode embedded in the laminated strip within the open path and proximate to the first end, a reagent matrix coextensive within the open path and covering the three electrodes, and conductive contacts located at the second end of the laminated strip.

Claims

1. A disposable electrode strip for testing a fluid sample comprising:

a laminated strip having a first strip end, a second strip end and a vent opening spaced from said first strip end, said laminated strip comprising a base layer having a conductive layer disposed thereon, said conductive layer having a scribe line delineated thereon and forming three electrode paths, a channel forming layer carried on said base layer, and a cover;

an enclosed channel between said first strip end and said vent opening, said enclosed channel sized to hold a volume of said fluid sample less than one microliter;

a reagent matrix containing at least an enzyme;

a stabilizer, wherein said stabilizer is a polyalkylene glycol; and

a redox mediator disposed on said base layer in said enclosed channel;

conductive contacts at said second strip end and insulated from said enclosed channel.

2. The electrode strip of claim 1 wherein said enzyme is glucose oxidase.

3. The electrode strip of claim 1 wherein said redox mediator is at least one metal complex.

4. The electrode strip of claim 3 wherein said redox mediator is at least one of potassium ferricyanide or other inorganic or organic redox mediators.

5. The electrode strip of claim 1 wherein said conductive coating is gold.

6. The electrode strip of claim 1 wherein said conductive coating comprising gold and tin oxide.

7. The electrode strip of claim 1 wherein said base layer, said channel forming layer and said cover are made of a plastic dielectric material.

8. The electrode strip of claim 1 wherein said enclosed channel is hydrophilic.

9. The electrode strip of claim 1 wherein said enclosed channel has a volume of about 0.22 microliters.

10. The electrode strip of claim 1 wherein said cover has a hydrophilic coating on at least one side.

11. The electrode strip of claim 1 wherein said reagent matrix further contains at least one of, a binder, a surfactant, and a buffer.

12. The electrode strip of claim 11 wherein, said binder is a cellulose material, and said surfactant is a polyoxyethylene ether.

13. The electrode strip of claim 12 wherein said stabilizer is polyethylene glycol, said binder is methyl cellulose, said surfactant is t-octylphenoxypolyethoxyethanol, and said buffer is a citrate buffer.

14. The electrode strip of claim 13 wherein said reagent matrix is made from a mixture having starting components comprising about 1 wt % to about 6.5 wt % of said redox mediator, about 2.5 wt % of said stabilizer, about 1 wt % of said binder, about 0.03 wt % of said surfactant, and about 1 wt % of said enzyme in said citrate buffer.

15. The electrode strip of claim 14 wherein said citrate buffer is about 0.05 M.

16. The electrode strip of claim 14 wherein said potassium ferricyanide is 6.5 wt %.

17. The electrode strip of claim 14 wherein said enzyme is glucose oxidase.

18. The electrode strip of claim 1 wherein said channel forming layer thickness sufficient to optimize the flow of said fluid sample along said open path.

19. The electrode strip of claim 18 wherein said thickness is about 0.0035 inches (0.089 mm).

20. The electrode strip of claim 1 wherein said enclosed channel contains a working electrode, a pseudo-working electrode and a reference electrode.

21. The electrode strip of claim 20 wherein said pseudo-working electrode is a counter electrode.

22. The electrode strip of claim 20 wherein said pseudo-working electrode is a triggering electrode.

23. The electrode strip of claim 20 wherein said pseudo-working electrode and said reference electrode pair are a resistance-measuring electrode pair.

24. The electrode strip of claim 1 wherein said stabilizer is polyethylene glycol.

25. A disposable electrode strip for detecting or measuring the concentration of an analyte in a fluid sample, said electrode strip comprising:

an insulating base strip having a first base end and a second base end;

a conductive layer disposed on one side of said base strip, said conductive layer having a pattern scribed into said conductive layer, said pattern delineating three electrically-distinct conductive paths insulated from each other;

a middle insulator sized smaller than said insulating base strip and overlaying a substantial portion of said conductive layer, said middle insulator having a cutout portion spaced from said first base end, said cutout portion exposing a limited area of said three conductive paths;

an electrode material comprising an enzyme, a redox mediator;

a stabilizer, wherein said stabilizer is a polyalkylene glycol, a binder, a surfactant, and a buffer, said electrode material being disposed in said cutout portion; and

a covering insulator sized to fit over and be coextensive with said middle insulator creating a sample fluid channel, said covering insulator having a covering insulator aperture spaced from said first base end and configured to expose at least a small portion of said cutout portion of said middle insulator.

26. The strip of claim 25 wherein said sample fluid channel has a volume of about 0.22 microliters.

27. The strip of claim 25 wherein said sample fluid channel is hydrophilic.

28. The strip of claim 25 wherein said redox mediator is at least one metal complex selected from the group consisting of ferrocene, ferrocene derivatives and potassium ferricyanide, said stabilizer is a polyalkylene glycol, said binder is a cellulose material, said surfactant is a polyoxyethylene ether, and said buffer has a pH of about 5 to about 6.

29. The strip of claim 28 wherein said mediator is potassium ferricyanide, said stabilizer is polyethylene glycol, said binder is methyl cellulose, said surfactant is t-octylphenoxypolyethoxyethanol, and said buffer is a citrate buffer.

30. The strip of claim 29 wherein said electrode material is made of a mixture having starting components comprising about 6.5 wt % of said potassium ferricyanide, about 2.5 wt % of said polyethylene glycol, about 1 wt % of said methyl cellulose, and about 0.03 wt % of said t-octylphenoxypolyethoxyethanol, and about 1 wt % of said enzyme in said citrate buffer.

31. The strip of claim 30 wherein said enzyme is glucose oxidase.

32. The electrode strip of claim 25 wherein said sample fluid channel contains a working electrode, a pseudo-working electrode and a reference electrode.

33. The electrode strip of claim 32 wherein said pseudo-working electrode is a counter electrode.

34. The electrode strip of claim 32 wherein said pseudo-working electrode is a triggering electrode.

35. The electrode strip of claim 32 wherein said pseudo-working electrode and said reference electrode pair are a resistance-measuring electrode pair.

36. A method of making multiple, disposable sensors wherein each sensor has at least a working electrode, a reference electrode, a pseudo-working electrode, and a reagent matrix, wherein said reagent matrix contains an enzyme capable of catalyzing a reaction involving a substrate for the enzyme, said working electrode and said reference electrode being disposed in a fluid sample channel for measuring a fluid sample, said method comprising:

obtaining a base strip of an insulating material having a layer of conductive material disposed thereon, said base strip having a first edge and a second edge;

scribing in said conductive material a plurality of lines in a repetitive pattern wherein said plurality of lines contain a repetitive pattern capable of forming three conductive paths in each of said repetitive pattern;

disposing a middle layer of insulating material over said base strip, said middle layer having a repetitive pattern of an elongated cutout wherein each cutout of each of said repetitive pattern exposes an electrode portion of each of said three conductive paths of each repetitive pattern wherein said repetitive pattern of said elongated cutout are spaced from said first edge of said base strip, and wherein said middle layer is sized to expose a contact portion of each of said two conductive paths of each repetitive pattern for a distance from said second edge of said base strip;

disposing an reagent material, wherein said reagent material contains a polyalkylene glycol stabilizer, into each elongated cutout of said repetitive pattern;

drying said reagent material at a temperature and for a length of time sufficient to solidify said reagent material in each of said elongated cutout;

disposing a top layer of insulating material over and coextensive with said middle layer, said top layer having a plurality of vent openings in a repetitive pattern wherein each of said vent openings exposes a portion of a corresponding repetitive pattern of said elongated cutout of said middle layer furthest from said first edge of said base strip, said base strip, said middle layer and said top layer forming a laminated strip;

cutting along and parallel to said first edge of said laminated strip a predetermined distance creating a sample inlet port in each of said elongated cutout for each of said repetitive pattern;

cutting along and parallel to said second edge of said laminated strip a predetermined distance creating two separate contacts for each of said repetitive pattern; and

separating each of said repetitive pattern at predetermined intervals along said laminated strip.

37. The method of claim 36 wherein said drying step further includes heating said reagent material at a temperature of about 55°

C.

38. The method of claim 37 wherein said drying step further includes heating said reagent material for about two minutes.