Small volume in vitro analyte sensor and methods of making

• US 8,377,378 B2
• Filed: 09/29/2009
• Issued: 02/19/2013
• Est. Priority Date: 10/08/1998
• Status: Expired due to Fees

• Current Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Inventors: Feldman, Benjamin J., Heller, Adam, Mao, Fei, Vivolo, Joseph A., Funderburk, Jeffery V., Colman, Fredric C., Krishnan, Rajesh, Heller, Ephraim
• Primary Examiner(s): LAM, ANN Y
• Application Number:
  US12/569,033
• Publication Number:
  US 20100018871A1
• Time in Patent Office: 1,239 Days
• Field of Search: None
• US Class Current: 422/68.1
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: G01N 27/3271 Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001
• 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 27/3274 Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration coding of calibration information G01N33/48771
• CPC Class Codes: Y10T 156/1052 with cutting, punching, tearing or severing
• CPC Class Codes: Y10T 29/49002 Electrical device making
• CPC Class Codes: Y10T 29/49004 including measuring or testing of device or component part
• CPC Class Codes: Y10T 29/49007 Indicating transducer
• CPC Class Codes: Y10T 29/49126 Assembling bases
• CPC Class Codes: Y10T 29/49128 Assembling formed circuit to base
• CPC Class Codes: Y10T 29/4913 Assembling to base an electrical component, e.g., capacitor, etc.
• CPC Class Codes: Y10T 29/49147 Assembling terminal to base
• CPC Class Codes: Y10T 29/49155 Manufacturing circuit on or in base
• CPC Class Codes: Y10T 29/49204 Contact or terminal manufacturing
• CPC Class Codes: Y10T 29/49208 by assembling plural parts
• CPC Class Codes: Y10T 29/4921 with bonding
• CPC Class Codes: Y10T 29/49794 Dividing on common outline

Abstract

A sensor utilizing a non-leachable or diffusible redox mediator is described. The sensor includes a sample chamber to hold a sample in electrolytic contact with a working electrode, and in at least some instances, the sensor also contains a non-leachable or a diffusible second electron transfer agent. The sensor and/or the methods used produce a sensor signal in response to the analyte that can be distinguished from a background signal caused by the mediator. The invention can be used to determine the concentration of a biomolecule, such as glucose or lactate, in a biological fluid, such as blood or serum, using techniques such as coulometry, amperometry, and potentiometry. An enzyme capable of catalyzing the electrooxidation or electroreduction of the biomolecule is typically provided as a second electron transfer agent.

Claims

1. A sensor for determining the concentration of an analyte in a sample fluid, the sensor comprising:

a first substrate having a proximal end and a distal end, the first substrate defining a first side edge and a second side edge of the electrochemical sensor extending from the proximal end to the distal end of the first substrate, the distal end being configured and arranged for insertion into a sensor reader;

a working electrode and a counter electrode, wherein the working electrode and counter electrode are separated by an effective distance in a range of 25 to 1000 μ

m, wherein the working electrode or the counter electrode is formed from a molded carbon fiber composite or comprises an inert non-conducting base material upon which a conducting layer is deposited selected from the group consisting of;

gold, carbon, platinum, ruthenium dioxide, palladium, and conductive epoxies;

a second substrate disposed over the first substrate, the working electrode being disposed on one of the first and second substrates and the counter electrode being disposed on one of the first and second substrates;

a spacer disposed between the first and second substrates and defining a first aperture and a second aperture, wherein the second aperture is defined along the second side edge;

a sample chamber extending from the first aperture to the second aperture for holding the sample fluid, the sample chamber comprising the working electrode and the counter electrode, wherein the sample chamber is sized to contain a volume of no more than about IL of the sample;

an analyte-responsive enzyme and a redox mediator disposed in the sample chamber; and

a first indicator electrode disposed on at least one of the first and second substrates and positioned to determine when the sample chamber contains sample.

2. The sensor of claim 1, wherein the sensor is configured and arranged such that the materials of the working and counter electrodes provide a background signal generated by the redox mediator that is no more than 5% of the signal generated by oxidation or reduction of the analyte.

3. The sensor of claim 1, wherein the first aperture is defined along the first side edge of the sensor.

4. The sensor of claim 1, wherein the first aperture is defined along the proximal end of the sensor.

5. The sensor of claim 1, wherein the indicator electrode is also a working electrode or a counter electrode.

6. The sensor of claim 1, further comprising a visual or auditory sign, coupled to the indicator electrode, that activates when the indicator electrode indicates that the sample chamber contains sample.

7. The sensor of claim 1, wherein the indicator electrode is disposed in facing relationship to one of the working electrode and the counter electrode.

8. The sensor of claim 1, wherein the sensor further comprises a second indicator electrode, wherein the first indicator electrode indicates when the sample chamber is beginning to fill with sample, and the second indicator electrode indicates when the sample chamber is substantially filled with sample.

9. The sensor of claim 8, wherein either the working electrode or the counter electrode is disposed between the first and second indicator electrodes.

10. The sensor of claim 1, wherein at least a portion of the working electrode is within an effective distance of no more than 200 μ

m of a portion of the counter electrode.

11. The sensor of claim 1, wherein the analyte is glucose and the analyte-responsive enzyme is a glucose-responsive enzyme.

12. The sensor of claim 1, wherein the analyte is glucose or ketone body.

13. The sensor of claim 1, wherein the sensor is configured and arranged so that the mediator oxidizes the analyte and the half-wave potential of the redox mediator, as measured by cyclic voltammetry in 0.1 M NaCl at pH 7, is no more than about +100 millivolts relative to the potential of the counter/reference electrode.

14. The sensor of claim 1, wherein the sensor is configured and arranged so that the mediator oxidizes the analyte and the half-wave potential of the redox mediator, as measured by cyclic voltammetry in 0.1 M NaCl at pH 7, is about the same as the potential of the counter/reference electrode.

15. The sensor of claim 1, wherein the sensor is configured and arranged so that the mediator oxidizes the analyte and the half-wave potential of the redox mediator, as measured by cyclic voltammetry in 0.1 M NaCl at pH 7, is no more than about −

150 millivolts relative to the potential of the counter/reference electrode.

16. The sensor of claim 1, wherein the redox mediator is diffusible, and wherein, within the sensor, the effective diffusion coefficient of the redox mediator through the sample fluid is less than the effective diffusion coefficient of the analyte through the sample fluid and, preferably, at least ten times less than the effective diffusion coefficient of the analyte through the sample fluid.

17. The sensor of claim 1, wherein the redox mediator is diffusible, and wherein the diffusible mediator has a molecular weight of at least 5,000 daltons.

18. The sensor of claim 1, wherein the redox mediator is diffusible, and wherein the sensor is configured and arranged so that the redox mediator is more readily electrolyzed on the working electrode than the counter electrode.

19. The sensor of claim 1, wherein the sensor comprises a molar amount of the redox mediator that is, on a stoichiometric basis, no more than an average normal physiological amount of the analyte and, preferably, the sensor comprises a molar amount of the redox mediator that is, on a stoichiometric basis, no more than 20% of an average normal physiological amount of the analyte.

20. The sensor of claim 1, wherein the working electrode has a surface area of no more than about 0.01 cm² exposed in the sample chamber.

21. The sensor of claim 1, wherein the activity of the enzyme is no more than 1 unit/cm³.

22. The sensor of claim 1, wherein the redox mediator is diffusible, and wherein the sensor is configured and arranged so that the diffusible redox mediator precipitates when reacted at the counter electrode.

23. The sensor of claim 1, wherein the redox mediator is diffusible, and wherein the sensor is configured and arranged so that a mathematical product of the effective diffusion coefficient of the redox mediator and the concentration of the redox mediator is no more than 1×

10<sup>−

12</sup> moles cm<sup>−

1</sup> sec<sup>−

1</sup> when sample fluid fills the sample chamber.

24. The sensor of claim 1, wherein the redox mediator is disposed on the working electrode.

25. The sensor of claim 1, wherein the analyte-responsive enzyme is disposed on the working electrode.

26. The sensor of claim 1, wherein the sample fluid is blood from the finger of a subject.

27. The sensor of claim 1, wherein the sample fluid is blood from a region of a subject having a lower nerve end density as compared to a fingertip.

28. The sensor of claim 27, wherein the region of a subject having a lower nerve end density as compared to a fingertip is selected from the group consisting of:

a forearm region, and a thigh region.

29. The sensor of claim 1, further comprising a sample acquisition apparatus.

30. The sensor of claim 29, wherein the sample acquisition apparatus comprises a lancet.

31. The sensor of claim 1, wherein the analyte is glucose and the analyte-responsive enzyme is selected from the group consisting of glucose dehydrogenase and glucose oxidase.

32. The sensor of claim 1, wherein the redox mediator comprises a transition metal complex.

33. The sensor of claim 32, wherein the transition metal complex comprises a transition metal selected from the group consisting of osmium, ruthenium, iron, and cobalt.

34. The sensor of claim 32, wherein the transition metal complex comprises osmium.

35. The sensor of claim 32, wherein the transition metal complex comprises two or more ligands coordinately bound to a transition metal.

36. The sensor of claim 35, wherein at least one of the ligands is a heterocyclic nitrogen-containing bidentate ligand.

37. The sensor of claim 35, wherein at least one of the ligands is a unsubstituted or substituted pyridine.

38. The sensor of claim 35, wherein at least one of the ligands is a halogen selected from the group consisting of fluorine, chlorine, and bromine.

39. The sensor of claim 1, wherein the sample chamber has a substantially uniform width between the first aperture and the second aperture that is not wider than either the first aperture or the second aperture.