Small volume in vitro analyte sensor with diffusible or non-leachable redox mediator

• US 6,299,757 B1
• Filed: 10/06/1999
• Issued: 10/09/2001
• Est. Priority Date: 10/08/1998
• Status: Expired due to Term

• Current Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories)
• Original Assignee: Therasense Incorporated (Abbott Laboratories)
• Inventors: Krishnan, Rajesh, Heller, Ephraim, Heller, Adam, Colman, Fredric C., Feldman, Benjamin J., Mao, Fei, Funderburk, Jeffery V., Vivolo, Joseph A.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/413,565
• Time in Patent Office: 734 Days
• Field of Search: 204/412, 204/409, 204/411, 204/403, 205/775, 205/777.5
• US Class Current: 205/775
• 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 a concentration of an analyte in a sample, the sensor comprising:

(a) 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 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;

(b) a second substrate disposed over the first substrate;

(c) a spacer disposed between the first and second substrates and defining a first aperture along the first side edge of the sensor, a second aperture along the second side edge of the sensor, and a sample chamber extending from the first aperture to the second aperture;

(d) at least one working electrode disposed on at least one of the first and second substrates;

(e) at least one counter electrode disposed on at least one of the first and second substrates, wherein the at least one working electrode and the at least one counter electrode are positioned relative to the sample chamber to generate an analyte-responsive signal when sample containing analyte is disposed in the sample chamber; and

(f) at least one indicator electrode disposed on at least one of the first and second substrates and positioned relative to a sample flow path within the sample chamber to indicate that, when sample contacts a one of the at least one indicator electrode, the sample chamber is filled with sample.

2. The sensor of claim 1, wherein portions of the first and second side edges proximate to the first and second apertures are straight.

3. The sensor of claim 1, wherein the sample chamber comprises two opposing straight edges extending between the first and second openings of the first and second side edges.

4. The sensor of claim 1, wherein the at least one indicator electrode is configured and arranged to aid in electrolysis of the analyte.

5. The sensor of claim 1, wherein the at least one indicator electrode comprises first and second indicator electrodes, the first indicator electrode is positioned relative to the sample chamber to determine when the sample chamber begins to fill with sample and the second indicator electrode is positioned relative to the sample chamber to determine when the sample chamber is filled with sample.

6. The sensor of claim 5, wherein a one of the at least one working electrodes and the at least one counter electrodes is positioned between the first and second indicator electrodes.

7. The sensor of claim 1, wherein the sample chamber comprises two opposing edges between the first and second side edges, the edges defining an expanded region in the sample chamber.

8. The sensor of claim 1, further comprising a diffusible redox mediator disposed in the sample chamber.

9. The sensor of claim 8, wherein the diffusible redox mediator is disposed on at least one of the at least one working electrodes.

10. The sensor of claim 8, wherein the sensor is configured and arranged so that a background signal generated by the diffusible redox mediator is no more than five times a signal generated by electrolysis of an average normal physiological amount of analyte.

11. The sensor of claim 1, further comprising a second electron transfer agent disposed in the sample chamber.

12. The sensor of claim 11, wherein the second electron transfer agent is disposed on at least one of the at least one working electrodes.

13. The sensor of claim 11, wherein the second electron transfer agent is diffusible.

14. The sensor of claim 4, wherein the at least one working electrode is disposed on the first substrate and extends over substantially all of the sample chamber and the at least one counter electrode is disposed on the second substrate.

15. The sensor of claim 4, wherein the at least one counter electrode is disposed on the second substrate and extends over substantially all of the sample chamber and the at least one working electrode is disposed on the first substrate.

16. The sensor of claim 1, wherein the sample chamber is sized to contain a volume of no more 1 μ

l of sample.

17. The sensor of claim 1, wherein the sample chamber is sized to contain a volume of no more than 0.5 μ

L of sample.

18. The sensor of claim 1, wherein the at least one working electrode is disposed on the first substrate, the at least one counter electrode is disposed on the second substrate, and the at least one working electrode is in a facing relationship to the at least one counter electrode and at least a portion of the working electrode is no more than 200 μ

m from a portion of the counter electrode.

19. The sensor of claim 1, wherein at least one of the at least one indicator electrode is configured and arranged for use as a counter electrode.

20. The sensor of claim 1, wherein at least one of the at least one indicator electrode is configured and arranged for use as a working electrode.

21. The sensor of claim 1, wherein the at least one indicator electrode is disposed on at least one of the first and second substrates and positioned, relative to a sample flow path within the sample chambers after at least one of the working and counter electrodes to indicate when the sanple chamber is sufficiently filled with sample.

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

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

(b) a second substrate disposed over the first substrate;

(c) a spacer disposed between the first and second substrates and defining a first aperture along the proximal end of the sensor, a second aperture along the first side edge of the sensor, and a sample chamber extending from the first aperture to the second aperture;

(d) at least one working electrode disposed on at least one of the first and second substrates;

(e) at least one counter electrode disposed on at least one of the first and second substrates, wherein the at least one working electrode and the at least one counter electrode are positioned relative to the sample chamber to generate an analyte-responsive signal when sample containing analyte is disposed in the sample chamber; and

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

23. The sensor of claim 22, wherein a portion of the first side edge proximate to the second aperture is straight.

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

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

(b) a second substrate disposed over the first substrate;

(c) a spacer disposed between the first and second substrates and defining a first aperture along the first side edge of the sensor, a second aperture along the second side edge of the sensor, and a sample chamber extending from the first aperture to the second aperture;

(d) at least one working electrode disposed on the first substrate;

(e) at least one counter electrode disposed on the second substrate, wherein the at least one working electrode and the at least one counter electrode are positioned relative to the sample chamber to generate an analyte-responsive signal when sample containing analyte is disposed in the sample chamber; and

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

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

(a) 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 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;

(b) a second substrate disposed over the first substrate;

(c) a spacer disposed between the first and second substrates and defining a first aperture along the first side edge of the sensor, a second aperture along the second side edge of the sensor, and a sample chamber extending from the first aperture to the second aperture;

(d) at least one working electrode disposed on at least one of the first and second substrates;

(e) at least one counter electrode disposed on at least one of the first and second substrates, wherein the at least one working electrode and the at least one counter electrode are positioned relative to the sample chamber to generate an analyte-responsive signal when sample containing analyte is disposed in the sample chamber; and

(f) at least one indicator electrode disposed on at least one of the first and second substrates wherein the at least one indicator electrode is configured and arranged to aid in electrolysis of the analyte.

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

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

(b) a second substrate disposed over the first substrate;

(c) a spacer disposed between the first and second substrates and defining a first aperture along the first side edge of the sensor, a second aperture along the second side edge of the sensor, and a sample chamber extending from the first aperture to the second aperture;

(d) at least one working electrode disposed on the first substrate;

(e) at least one counter electrode disposed on the second substrate, wherein the at least one working electrode and the at least one counter electrode are positioned relative to the sample chamber to generate an analyte-responsive signal when sample containing analyte is disposed in the sample chamber; and

(f) at least one indicator electrode disposed on at least one of the first and second substrates, wherein the at least one indicator electrode comprises first and second indicator electrodes, the first indicator electrode is positioned relative to the sample chamber to determine when the sample chamber begins to fill with sample and the second indicator electrode is positioned relative to the sample chamber to determine when the sample chamber is filled with sample.

27. A method of monitoring an analyte an electrochemical sensor, the method comprising:

contacting a sample with an aperture defined between two substrates of an electrochemnical scraper comprising at least one working electrode disposed on a one of the two substrates;

drawing at least a portion of the sample through the aperture and into a sample chamber disposed between the two substrates;

observing an indicator electrode disposed on a one of the two substrates, wherein the indicator electrode is positioned within the sensor to indicate, when the sample contacts the working electrode and then subsequently contacts the indicator electrode, that the sample chamber is sufficiently full for monitoring the analyte using the sensor; and

generating an analyte-responsive signal using the at least one working electrode and at least one counter electrode disposed on a one of the two substrates, wherein the at least one working electrode and at least one counter electrode are separate from the indicator electrode and are disposed in electrolytic contact with the sample chamber.

28. The method of claim 27, further comprising

observing a second indicator electrode disposed on a one of the two substrates, wherein the second indicator electrode is positioned within the sensor to indicate, when sample contact the indicator electrode, that the sample chamber is beginning to fill with sample.

29. The method of claim 27, wherein generating an analyte-responsive signal comprises generating the analyte-responsive signal using the at least one working electrode, the at least one counter electrode, and the indicator electrode.

30. The method of claim 27, wherein the step of generating an analyte-responsive signal occurs simultaneously with and continues after the step of observing the indicator electrode.

31. The method of claim 28, wherein the step of generating an analyte responsive signal occurs after the step of observing the indicator electrode.