Electrochemical assay device and related methods

• US 8,617,365 B2
• Filed: 05/21/2005
• Issued: 12/31/2013
• Est. Priority Date: 05/21/2004
• 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
• Primary Examiner(s): DINH, BACH T
• Application Number:
  US10/908,664
• Publication Number:
  US 20050258036A1
• Time in Patent Office: 3,146 Days
• Field of Search: 20440301-40315, 2057775-778
• US Class Current: 204/403.01
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: G01N 27/30 Electrodes, e.g. test electrodes; Half-cells G01N27/414 takes precedence
• CPC Class Codes: G01N 27/3272 Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels optical biosensors G01N33/52
• CPC Class Codes: G01N 27/403 Cells and electrode assemblies
• CPC Class Codes: Y10T 156/1052 with cutting, punching, tearing or severing
• CPC Class Codes: Y10T 156/1062 Prior to assembly
• CPC Class Codes: Y10T 156/1074 Separate cutting of separate sheets or webs
• CPC Class Codes: Y10T 29/49002 Electrical device making
• CPC Class Codes: Y10T 29/49114 including adhesively bonding
• CPC Class Codes: Y10T 29/49117 Conductor or circuit manufacturing

Abstract

An electrochemical test device is provided having a base layer with a first electrode thereon and a top layer with a second electrode thereon. The two electrodes are separated by a spacer layer having an opening therein, such that a sample-receiving space is defined with one electrode on the top surface, the other electrodes on the bottom surface and side walls formed from edges of the opening in the spacer. Reagents for performing the electrochemical reaction are deposited on one of the electrodes and on the side walls of the sample-receiving space.

Claims

1. An electrochemical test device comprising

a bottom substrate having a first electrode disposed thereon,

a top substrate having a second electrode disposed thereon,

a spacer disposed between the top and bottom substrates and having an opening therein, whereby a sample-receiving space is defined that has a bottom surface having the first electrode disposed thereon, a top surface, opposite the bottom surface, having the second electrode disposed thereon, and side walls formed from edges of the opening in the spacer;

and

a reagent comprising a redox active material which disperses in a liquid sample placed in the sample-receiving space and is oxidized at the first electrode and reduced at the second electrode when the device is used;

wherein in the test device prior to introduction of a liquid sample, the reagent is disposed in a layer, a first part of said layer being disposed on and covering at least a portion of the first or second electrode and a second part of said layer being disposed upon and covering at least a portion of the side walls, wherein said second part of said layer extends on the side walls away from the first part of said layer such that the layer as a whole has a U-shaped cross section.

2. The device of claim 1, wherein the second part of the layer of reagent covers at least 25% of the height of the side walls.

3. The device of claim 1, wherein the second part of the layer of reagent covers at least 50% of the height of the side walls.

4. The device of claim 1, wherein the second part of the layer of reagent covers at least 75% of the height of the side walls.

5. The device of claim 1, wherein the second part of the layer of reagent covers 100% of the height of the side walls.

6. The device of claim 1, wherein the redox active material is selected from the group consisting of ferricyanide, [FeIII(CN)5(ImH)]²⁻

, [FeIII(CN)5(Im)]³⁻

, [RuIII(NH3)5(ImH)]³⁺, [RuIII(NH3)5(Im)]²⁺, [FeII(CN)5(ImH)]³⁻

, [RuII(NH3)5(Im)H]²⁺, [(NC)5FeII(Im)RuIII(NH3)5]⁻

, [(NC)5FeIII(Im)RuIII(NH3)5]⁰, [(NC)5FeII(Im)RuII(NH3)5]²⁻

, Ferrocene (Fc), Ferrocene monosulphonate, Ferrocene disulphonate, FcCO₂H, FcCH2CO₂H, FcCH;

CHCO₂H, Fc(CH₂)₃CO₂H, Fc(CH₂)₄CO₂H, FcCH₂CH(NH₂)CO₂H, FcCH₂SCH₂CH(NH₂)CO₂H, FcCH₂CONH₂, Fc(CH₂)₂CONH₂, Fc(CH₂)₃CONH₂, Fc(CH₂)₄CONH₂, FcOH, FcCH₂OH, Fc(CH₂)₂OH, FcCH(Me)OH, FcCH₂O(CH₂)₂OH, 1,1′

-Fc(CH₂OH)₂, 1,2-Fc(CH₂OH)₂, FcNH₂, FcCH₂NH₂, Fc(CH₂)₂NH₂, Fc(CH₂)₃NH₂, 1,1′

-Me₂FcCH₂NH₂, FcCH₂NMe₂, (R)-FcCH(Me)NMe₂, (S)-FcCH(Me)NMe₂, 1,2-Me₃SiFcCH₂NMe₂, FcCH₂NMe₃, FcCH₂NH(CH₂)₂NH₂, 1,1′

-Me₂FcCH(OH)CH₂NH₂, FcCH(OH)CH₂NH₂, FcCH;

CHCH(OH)CH₂NH₂, Fc(CH₂)₂CH(OH)CH₂NH₂, FcCH₂CH(NH₂)CH₂OH, FcCH₂CH(CH₂NH₂)CH₂OH, FcCH₂NH(CH₂)₂OH, 1,1′

-Me₂FcCHOCONHCH₂, FcCH(OH)(CH₂)₂NH₂, 1,1′

-Me₂FcCH(OH)CH₂NHAc, FcB(OH)₃, FcC₆H₄OPO₃Na₂, Os(4,7-dmphen)₃, Os(3,4,7,8-tmphen)₃, Os(5,6-dmphen)₃, Os(bpy)₃Cl₂, Os(5-mphen)₃, Os(5-Cl-phen)₃, Os(5-NO₂-phen)₃, Os(5-phphen)₃, Os(2,9-dm-4,7-dpphen)₃, Ru(4,7-dmphen)₃, Ru(3,4,7,8-tmphen)₃, Ru(5-mphen)₃, Ru(5,6-dmphen)₃, Ru(phen)₃, [Ru(4,4′

-diNH₂-bipy)3]²⁺, Os(bpy)₃, Os(dmbpy)₃, Ru(bpy)₃, Ru(4,4′

-diNH₂-bpy)₃, Ru(4,4′

-diCO₂Etbpy)₃, Os(bpy)₂dmbpy, Os(bpy)₂(HIm)₂, Os(bpy)₂(2MeHIm)₂, Os(bpy)₂(4MeHIm)₂, Os(dmbpy)₂(HIm)₂, Os(bpy)₂Cl(HIm), Os(bpy)₂Cl(1-MeIm), Os(dmbpy)₂Cl(HIm), Os(dmbpy)₂Cl(1-MeIm), Ru(bpy)₂(5,5′

diNH₂-bpy), Ru(bpy)₂(5,5′

diCO₂Etbpy), Ru(bpy)₂(4,4′

diCO₂Etbpy), or the complementary redox forms (oxidized or reduced) thereof.

7. The device of claim 6, wherein the reagent further comprises glucose oxidase.

8. The device of claim 7, wherein the second part of the layer of reagent covers at least 25% of the height of the side walls.

9. The device of claim 7, wherein the second part of the layer of reagent covers at least 50% of the height of the side walls.

10. The device of claim 7, wherein the second part of the layer of reagent covers at least 75% of the height of the side walls.

11. The device of claim 7, wherein the second part of the layer of reagent covers 100% of the height of the side walls.

12. The device of claim 6, wherein the second part of the layer of reagent covers at least 25% of the height of the side walls.

13. The device of claim 6, wherein the second part of the layer of reagent covers at least 50% of the height of the side walls.

14. The device of claim 6, wherein the second part of the layer of reagent covers at least 75% of the height of the side walls.

15. The device of claim 6, wherein the second part of the layer of reagent covers 100% of the height of the side walls.

16. A method of making an electrochemical test device comprising the steps of:

(a) forming a bottom substrate having a first electrode disposed thereon;

(b) forming a spacer layer on the bottom substrate, said spacer layer having an opening formed therein through which the first electrode is exposed and side walls within the opening;

(c) introducing a liquid reagent comprising a redox active material into the opening in the spacer layer,

(d) drying the liquid reagent to form a dried reagent layer, wherein the liquid reagent is introduced to the opening in such a manner that upon drying a dried reagent layer is formed, said dried reagent layer having a first part covering at least a portion of the first electrode and a second part covering at least a portion of the side walls, wherein said second part of said layer extends on the side walls away from the first part of said layer such that the layer as a whole has a U-shaped cross section; and

(e) placing a top substrate having a second electrode disposed thereon on the spacer layer aligned to have the second electrode facing the first electrode, thereby forming a sample-receiving space that has a first surface having the first electrode disposed thereon, a second surface, opposite the first surface, having the second electrode disposed thereon, and side walls formed from edges of the opening in the spacer.

17. The method of claim 16, wherein the second part of the layer of dried reagent covers at least 25% of the height of the side walls.

18. The method of claim 16, wherein the second part of the layer of dried reagent covers at least 50% of the height of the side walls.

19. The method of claim 16, wherein the second part of the layer of dried reagent covers at least 75% of the height of the side walls.

20. The method of claim 16, wherein the second part of the layer of dried reagent covers 100% of the height of the side walls.

21. The method of claim 16, wherein the spacer layer applied in step (b) comprises an adhesive coating and a release sheet disposed on the side of the spacer layer facing away from the first electrode, wherein an edge of the release sheet is aligned with the sidewall thereby increasing the height of the sidewall further comprising the step of removing the release sheet after drying of the liquid reagent to expose the adhesive layer.

22. The method of claim 21, wherein the liquid reagent is introduced to the opening in a volume sufficient to fill the opening to a level that at least partially covers the portion of the side walls formed from the release sheet.

23. The method of claim 16, wherein the liquid reagent is introduced in a volume sufficient to fill the opening to a level that partially covers the portion of the side walls of the opening.

24. The method of claim 16, wherein the opening and the bottom substrate form a well that is bounded by side walls on all sides.

25. The method of claim 16, wherein the opening and the bottom substrate form a channel bounded by side walls on only two opposing sides.