Immunosensor

• US 8,685,714 B2
• Filed: 04/22/2004
• Issued: 04/01/2014
• Est. Priority Date: 07/14/2000
• Status: Expired due to Term

• Current Assignee: Universal Biosensors Pty Ltd. (Universal Biosensors, Inc.)
• Original Assignee: Universal Biosensors Pty Ltd. (Universal Biosensors, Inc.)
• Inventors: Hodges, Alastair, Chatelier, Ron
• Primary Examiner(s): Brown, Melanie Y
• Assistant Examiner(s): Crawford, Erik B
• Application Number:
  US10/830,841
• Publication Number:
  US 20040203137A1
• Time in Patent Office: 3,631 Days
• Field of Search: 435/4, 435/7.1, 435/7.92, 435174-177, 435/287.1, 435/287.2, 435/288.5, 435/288.7, 435/808, 435/7.91, 435/14, 435/190, 435/19, 435/2, 435/288.4, 435/966, 435/7.93, 436518-528, 436/164, 436/172, 436/805, 436/806, 436/904, 436/809, 422/68.1, 422/82.01, 422/82.05, 422/947
• US Class Current: 435/288.5
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: C12Q 1/006 for glucose
• 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 33/84 involving inorganic compounds or pH

Abstract

This invention describes a quantitative, inexpensive, disposable immunosensor that requires no wash steps and thus generates no liquid waste. Moreover, in preferred embodiments of the sensor no timing steps are required of the user, and the sensor can be readily adapted to antigen-antibody interactions over a wide kinetic range.

Claims

1. An electrochemical device for use in detecting a target antigen in a fluid sample, the device comprising:

a reaction chamber including

a) an internal surface, a proximal end, and a distal end,

b) an immobilized antibody that can bind to the target antigen, said antibody being immobilized to an internal surface of the reaction chamber or said antibody being immobilized onto a support placed within the reaction chamber, and

c) a reporter comprising an enzyme probe and an antigen configured for binding with the immobilized antibody;

a detection chamber including

a) an internal surface, a distal end, and a proximal end,

b) electrodes for detecting an electrochemical reaction in the detection chamber, and

c) a zone having a wall at the distal end of the detection chamber, wherein the wall is capable of being pierced to form a vent; and

a sample passageway between the distal end of the reaction chamber and the proximal end of the detection chamber, the detection chamber trapping air when the fluid sample enters the sample passageway, preventing ingress of the fluid sample into the detection chamber;

wherein the device is adapted to move the sample from the reaction chamber to the detection chamber via capillary action upon opening the vent by piercing the wall of the zone in the detection chamber to release trapped air, and

wherein the presence of a target antigen in the fluid sample results in a measurable change in an electrochemical reaction in the detection chamber.

2. The device of claim 1, wherein the reaction chamber includes an excess of antibody.

3. The device of claim 1, wherein the enzyme comprises glucose oxidase.

4. The device of claim 1, wherein the enzyme comprises glucose dehydrogenase.

5. The device of claim 1, wherein the detection chamber further comprises an enzyme substrate.

6. The device of claim 5, wherein the enzyme substrate is an oxidizable enzyme substrate.

7. The device of claim 6, wherein the oxidizable enzyme substrate is selected from the group consisting of galactose and acetic acid.

8. The device of claim 6, wherein the oxidizable enzyme substrate comprises glucose.

9. The device of claim 1, wherein the detection chamber further comprises a mediator.

10. The device of claim 9, wherein the mediator is selected from the group consisting of dichlorophenolindophenol and complexes between the transition metals and nitrogen-containing heteroatomic species.

11. The device of claim 9, wherein the mediator comprises ferricyanide.

12. The device of claim 1, wherein the detection chamber further comprises a buffer for adjusting the pH of a sample.

13. The device of claim 12, wherein the buffer comprises phosphate.

14. The device of claim 12, wherein the buffer comprises citrate.

15. The device of claim 1, further comprising an agent contained within the reaction chamber, the agent adapted to prevent non-specific binding of proteins to the reaction chamber internal surface.

16. The device of claim 15, wherein the agent is selected from the group consisting of a surfactant and a blocking protein.

17. The device of claim 15, wherein the agent is bovine serum albumin.

18. The device of claim 1, wherein the reporter is spaced from the immobilized antibody by less than about 1 millimeter.

19. The device of claim 1, wherein the device further comprises a support material.

20. The device of claim 19, wherein the support material is positioned within the detection chamber and wherein a first substance selected from the group consisting of an enzyme substrate, a mediator, a buffer, and combinations thereof is supported on or contained within the support material.

21. The device of claim 19, wherein the support material is positioned within the reaction chamber and wherein a second substance selected from the group consisting of the immobilized antibody, the reporter, and an agent adapted to prevent non-specific binding of proteins to the reaction chamber internal surface is supported on or contained within the support material.

22. The device of claim 19, wherein the support material comprises a mesh material.

23. The device of claim 19, wherein the support material comprises a fibrous filling material.

24. The device of claim 19, wherein the support material comprises a porous material.

25. The device of claim 24, wherein the porous material comprises sintered powder.

26. The device of claim 24, wherein the porous material comprises a macroporous membrane.

27. The device of claim 1, further comprising a detector adapted to detect a condition wherein the reaction chamber is substantially filled.

28. The device of claim 1, further comprising a reaction chamber vent at the distal end of the reaction chamber.