Small volume in vitro analyte sensor

• US 6,551,494 B1
• Filed: 04/06/2000
• Issued: 04/22/2003
• Est. Priority Date: 02/06/1997
• Status: Expired due to Term

• Current Assignee: Abbott Diabetes Care Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Therasense Incorporated (Abbott Laboratories Incorporated)
• Inventors: Say, James, Heller, Adam, Vreeke, Mark S., Feldman, Benjamin J.
• Primary Examiner(s): Tung, T.
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/544,593
• Time in Patent Office: 1,111 Days
• Field of Search: 205/777.5, 205/792, 204/403.14, 422/8.05, 600/583, 600/347
• US Class Current: 205/777.5
• CPC Class Codes: A61B 2560/0223 of calibration, e.g. protocols for calibrating sensors
• CPC Class Codes: A61B 2562/0295 Strip shaped analyte sensors for apparatus classified in A61B5/145 or A61B5/157 disposable multilayered or laminated electrodes G01N27/307; details of handling chemical test elements G01N33/4875
• CPC Class Codes: A61B 5/1411 by percutaneous method, e.g. by lancet
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/14546 for measuring analytes not otherwise provided for, e.g. ions, cytochromes
• CPC Class Codes: A61B 5/1486 using enzyme electrodes, e.g. with immobilised oxidase
• CPC Class Codes: A61B 5/14865 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
• CPC Class Codes: A61B 5/1495 Calibrating or testing of in-vivo probes
• CPC Class Codes: A61B 5/150022 for capillary blood or interstitial fluid
• CPC Class Codes: A61B 5/150358 Strips for collecting blood, e.g. absorbent optical reagent test strips G01N21/8483; chemical reagent test strips G01N33/4875, G01N33/52
• CPC Class Codes: A61B 5/150412 Pointed piercing elements, e.g. needles, lancets for piercing the skin blade-like piercing elements A61B5/150442
• CPC Class Codes: A61B 5/150503 Single-ended needles hollow piercing element tip design A61B5/150396, needle sets for catheters A61M25/06, needles for bringing media into the body A61M5/32
• CPC Class Codes: A61B 5/15105 Purely manual piercing, i.e. the user pierces the skin without the assistance of any driving means or driving devices
• CPC Class Codes: A61B 5/15113 Manually triggered, i.e. the triggering requires a deliberate action by the user such as pressing a drive button
• CPC Class Codes: A61B 5/15144 comprising driving means, e.g. a spring, for retracting the piercing unit into the housing
• CPC Class Codes: A61B 5/157 Devices characterised by integrated means for measuring characteristics of blood
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: G01N 2333/904 acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
• 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 27/3273 : Devices therefor, e.g. test element readers, circuitry details not specific to biochemical electrodes G01N33/4875
• CPC Class Codes: G01N 33/5438 : Electrodes

Abstract

A sensor designed to determine the amount and concentration of analyte in a sample having a volume of less than about 1 μL. The sensor has a working electrode coated with a non-leachable redox mediator. The redox mediator acts as an electron transfer agent between the analyte and the electrode. In addition, a second electron transfer agent, such as an enzyme, can be added to facilitate the electrooxidation or electroreduction of the analyte. The redox mediator is typically a redox compound bound to a polymer. The preferred redox mediators are air-oxidizable.

The amount of analyte can be determined by coulometry. One particular coulometric technique includes the measurement of the current between the working electrode and a counter or reference electrode at two or more times. The charge passed by this current to or from the analyte is correlated with the amount of analyte in the sample. Other electrochemical detection methods, such as amperometric, voltammetric, and potentiometric techniques, can also be used.

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. An enzyme capable of catalyzing the electrooxidation or electroreduction of the biomolecule is provided as a second electron transfer agent.

Claims

1. A method for determining a concentration of glucose in blood, comprising the steps of:

piercing the forearm of a patient to draw a blood sample of about 1 microliter or less of blood, wherein piercing is performed by piercing with an analyte measurement device comprising an electrochemical sensor and a piercing member adapted to pierce the skin and draw blood;

directly transferring about 1 microliter or less of the blood sample to the electrochemical sensor, the electrochemical sensor comprising;

a first non-conductive substrate and a second non-conductive substrate each having parallel side edges and ends, the substrates held together with an adhesive,

a sample chamber between the substrates having a thickness of less than about 0.2 mm, the sample chamber comprising a measurement zone sized to contain less than about 1 microliter and including a glucose-responsive enzyme,

a working electrode positioned on one of the substrates, and

a counter electrode positioned on one of the substrates;

filling the measurement zone with at least a portion of the blood sample;

holding the sample within the measurement zone in a non-flowing manner; and

determining the concentration of glucose in the sample using a coulometric technique.

2. The method of claim 1, wherein the sample chamber holds the sample in electrolytic contact with the working electrode.

3. The method of claim 2 wherein the sensor further comprises a reference electrode, and the sample chamber holds the sample in electrolytic contact with the working electrode, the counter electrode and the reference electrode.

4. The method of claim 2, wherein the working electrode and the counter electrode are a facing electrode pair.

5. The method of claim 2, wherein the glucose-responsive enzyme is non-leachably disposed on the working electrode.

6. The method of claim 2, wherein the glucose-responsive enzyme is immobilized on the working electrode.

7. The method of claim 2, wherein the sensor further comprises a redox mediator in the measurement zone.

8. The method of claim 2, wherein the sensor further comprises a redox mediator disposed on the working electrode.

9. The method of claim 8, wherein the redox mediator is immobilized on the working electrode.

10. The method of claim 8, wherein the redox mediator is non-leachable.

11. The method of claim 1, further comprising positioning the sensor before the forearm of the patient is pierced.

12. The method of claim 1, wherein directly transferring about 1 microliter or less of the sample to the electrochemical sensor comprises transferring to the electrochemical sensor comprising a measurement zone sized to contain less than about 0.5 microliter.

13. The method of claim 1, wherein directly transferring about 1 microliter or less of the sample to the electrochemical sensor comprises transferring to the electrodchemical sensor by capillary action.

31. The method of claim 1, wherein directly transferring about 1 microliter or less of the sample to the electrochemical sensor comprises transferring to the electrochemical sensor comprising a sample chamber having thickness of less than about 0.1 mm.

14. A method for determining a concentration of glucose in blood of a patient, comprising the steps of:

creating an unassisted flow of less than about 1 microliter of blood from the forearm of the patient, wherein creating an unassisted flow is performed by piercing with an analyte measurement device comprising an electrochemical sensor and a piercing member adapted to pierce the skin and draw blood;

directly transferring a portion of the blood into the electrochemical glucose sensor configured and arranged to determine the concentration of glucose from 1 microliter or less of blood, the sensor comprising;

a first non-conductive substrate and a second non-conductive substrate each having parallel side edges and ends, the substrates held together with an adhesive,

a sample chamber between the substrates having a thickness of less than about 0.2 mm, the sample chamber comprising a measurement zone sized to contain less than about 1 microliter and including a glucose-responsive enzyme,

a working electrode positioned on one of the substrates, and

a counter electrode positioned on one of the substrates;

holding the blood within the measurement zone in a non-flowing manner; and

determining the concentration of the glucose in the blood from the portion of the blood transported into the measurement zone.

15. The method of claim 14, wherein the sample chamber holds the blood in electrolytic contact with the working electrode.

16. The method of claim 14, wherein the electrochemical sensor further comprises a reference electrode, and the sample chamber holds the sample in electrolytic contact with the working electrode, the counter electrode and the reference electrode.

17. The method of claim 15, wherein the working electrode and the counter electrode are a facing electrode pair.

18. The method of claim 14, wherein the glucose-responsive enzyme is disposed on the working electrode.

19. The method of claim 14, wherein the sensor comprises a non-leachable glucose-responsive enzyme disposed on the working electrode.

20. The method of claim 14, wherein the glucose-responsive enzyme is immobilized on the working electrode.

21. The method of claim 14, wherein the sensor further comprises a redox mediator disposed on the working electrode.

22. The method of claim 21, wherein the redox mediator comprises a non-leachable redox mediator disposed on the working electrode.

23. The method of claim 21, wherein the redox mediator is immobilized on the working electrode.

24. The method of claim 14, wherein determining the concentration of the glucose comprises determining the concentration of the glucose using a coulometric technique.

25. The method of claim 14, wherein determining the concentration of the glucose comprises determining the concentration of the glucose using an amperometric technique.

26. The method of claim 14, wherein directly transferring a portion of the blood into the sensor comprises wicking a portion of the blood into the sample chamber with a sorbent material disposed in the sample chamber.

27. The method of claim 14, wherein directly transferring a portion of the blood into the sensor comprises transferring by capillary action.

28. The method of claim 14, wherein directly transferring a portion of the blood into the sensor comprises directly transferring by application of a vacuum.

29. The method of claim 14, wherein directly transferring a portion of the blood into the sensor comprises transferring a portion of the blood into the sensor comprising a sample chamber having a thickness of less than about 0.1 mm.

30. The method of claim 14, wherein directly transferring a portion of the blood into the sensor comprises transferring a portion of the blood into the sensor comprising a measurement zone sided to contain less than about 0.5 microliter.

32. A method for determining the concentration of glucose in blood, the method comprising the steps of:

piercing the skin of a forearm with a lancet to draw 1 microliter or less of blood, wherein piercing is performed by piercing with an analyte measurement device comprising an electrochemical sensor and a piercing member adapted to pierce the skin and draw blood;

directly transferring the blood to the electrochemical sensor and into a sample chamber in the sensor, the sensor comprising;

two plates each having parallel side edges and ends, the plates being formed of non-conducting material held together with an adhesive, the sample chamber having a thickness between the plates of less than about 0.2 mm, and

positioned on the plates, a working electrode and a counter electrode, the sample chamber including a measurement zone between the working electrode and the counter electrode, the measurement zone being sized to contain less than about 0.5 microliter of the blood;

permitting the blood to come to rest in the sample chamber;

determining the concentration of glucose in the blood using an amperometric technique.

33. The method of claim 32, wherein the step of directly transferring the blood to the sensor comprises directly transferring the blood to an end of the plates.

34. A method for determining the concentration of glucose in blood, the method comprising the steps of:

piercing the skin of a forearm with a lancet to draw 1 microliter or less of blood, wherein piercing is performed by piercing with an analyte measurement device comprising an electrochemical sensor and a piercing member adapted to pierce the skin and draw blood;

directly transferring the blood to the electrochemical sensor and into a sample chamber in the sensor, the sensor comprising;

two plates each having parallel side edges and ends, the plates being formed of non-conducting material held together with an adhesive, the sample chamber having a thickness between the plates of less than about 0.2 mm, and

positioned on the plates, a working electrode and a counter electrode, the sample chamber including a measurement zone between the working electrode and the counter electrode, the measurement zone being sized to contain less than about 0.5 microliter of the blood;

permitting the blood to come to rest in the sample chamber;

determining the concentration of glucose in the blood using a coulometric technique.

35. The method of claim 34, wherein the step of directly transferring the blood to the sensor comprises directly transferring the blood to an end of the plates.

36. A method for determining the concentration of glucose in blood using an analyte measuring device comprising a piercing member and an electrochemical sensor, the method comprising:

piercing the skin of the forearm of a patient to draw a blood sample of about 1 microliter or less, wherein piercing is performed by piercing the skin using the piercing member of the analyte measuring device;

directly transferring about 1 microliter or less of the blood sample to the electrochemical sensor, the electrochemical sensor comprising;

a first non-conductive substrate and a second non-conductive substrate, the substrates held together with an adhesive,

a sample chamber between the substrates having a thickness of less than about 0.2 mm, the sample chamber comprising a measurement zone sized to contain less than about 1 microliter of the sample and including a glucose responsive enzyme,

a working electrode on one of the substrates,

a counter electrode positioned on one of the substrates;

filling the measurement zone with at least a portion of the sample;

holding the sample within the measurement zone in a non-flowing manner; and

determining the concentration of glucose in the sample using a coulometric or amperometric approach.

37. The method of claim 36, wherein directly transferring a portion of blood into the sensor comprises applying a vacuum.