Biosensor Apparatus and Methods of Use

• US 20100006452A1
• Filed: 09/18/2009
• Published: 01/14/2010
• Est. Priority Date: 03/21/2002
• Status: Active Grant

• Current Assignee: Universal Biosensors Pty Ltd. (Universal Biosensors, Inc.)
• Original Assignee: Universal Biosensors Pty Ltd. (Universal Biosensors, Inc.)
• Inventors: Chatelier, Ronald, Hodges, Alastair McIndoe, Rylatt, Dennis
• Granted Patent: US 9,863,942 B2
• Time in Patent Office: Days
• US Class Current: 205/777.500
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: Y10T 29/49124 On flat or curved insulated base, e.g., printed circuit, etc.

Abstract

Disclosed herein are methods and devices for detecting the presence of an analyte of interest. A biosensor device can include a reaction chamber and an electrochemical detection chamber. The reaction chamber can include at least one immobilized binding site and a probe conjugate adapted to bind to at least one of the target analyte and the immobilized binding site while the detection chamber can include electrodes for detecting an electrochemical reaction. If present, the target analyte in the fluid sample results in a change in the amount of probe conjugate bound in the reaction chamber, which can be detected electrochemically in the detection chamber.

Claims

1-24. -24. (canceled)

25. A biosensor for use in detecting a target analyte in a fluid sample, the device comprising:

a reaction chamber including an immobilized binding site and a probe conjugate, wherein the probe conjugate comprises a binding partner adapted to bind to the immobilized binding site, wherein the immobilized binding site is positioned on at least one magnetic bead, wherein the immobilized binding site and the probe conjugate are positioned on different surfaces of the reaction chamber;

a detection chamber including electrodes for detecting an electrochemical reaction in the detection chamber; and

a fluid passageway between the reaction chamber and the detection chamber,

wherein, the presence or absence of a target analyte in the fluid sample results in a change in the amount of probe conjugate bound in the reaction chamber, the change detectable with an electrochemical reaction in the detection chamber.

26. The biosensor of claim 25, wherein the immobilized binding site is an antigen and the binding partner is an antibody.

27. The biosensor of claim 25, wherein the immobilized binding site is an antibody and the binding partner is an antigen.

28. The biosensor of claim 25, further comprising a filling chamber adapted to be filled by capillary action.

29. The biosensor of claim 28, wherein the capillary dimension of the reaction chamber is smaller in size than that of the filling chamber.

30. The biosensor of claim 25, wherein the detection chamber includes a vent.

31. The biosensor of claim 30, wherein the vent can be opened by piercing an outer layer of the device.

32. The biosensor of claim 30, wherein the vent can be opened by removing a portion of an outer layer of the device.

33. The biosensor of claim 30, wherein the vent can be opened by tearing along a perforation.

34. The biosensor of claim 25, wherein the reaction chamber includes an opening to the atmosphere.

35. The biosensor of claim 25, further comprising a magnet.

36. The biosensor of claim 35, wherein the magnet is disposed so as to move the at least one magnetic bead to be in intimate contact with the probe conjugate after the fluid sample has been introduced into the reaction chamber.

37. The biosensor of claim 25, wherein the probe conjugate comprises an enzyme.

38. The biosensor of claim 37, further comprising a mediator and an enzyme substrate.

39. A method of detecting a target analyte in a fluid sample, the method comprising the steps of:

delivering a sample to the biosensor of claim 25;

allowing a reaction to proceed in the reaction chamber between an immobilized binding site and a probe conjugate; and

moving the sample into the detection chamber and electrochemically detecting the level of probe conjugate, wherein the presence of the target analyte in the sample results in an increase or a decrease in the amount of probe conjugate detected in the detection chamber.

40. The method of claim 39, wherein the immobilized binding site includes a target antigen and the probe conjugate includes an antibody adapted to bind to the target antigen.

41. The method of claim 39, wherein the probe conjugate includes an enzyme.

42. The method of claim 39, wherein the sample is moved from the reaction chamber to the detection chamber via capillary action.

43. The method of claim 39, wherein the step of moving the sample includes opening a vent.

44. The method of claim 39, further comprising the step of quantifying the amount of target antigen in the sample based electrical signals received from the detection chamber.

45. A method of manufacture of the biosensor of claim 28, the method comprising the steps of:

providing a first layer;

removing a section of the first layer to form a filling chamber part, a reaction chamber part, and a detection chamber part;

providing a second layer;

mounting a first side of the second layer to a first side of the first layer;

removing a section of the second layer extending over the filling chamber part, in substantial registration with the filling chamber part, the second layer further extending over the reaction chamber part and the detection chamber part;

providing a third layer;

mounting a first side of the third layer to a second side of the first layer, the third layer extending over the filling chamber part, the reaction chamber part and the detection chamber part;

mounting a first side of a sealing layer to a second side of the second layer, whereby the layers comprising the first layer, the second layer, the third layer and the sealing layer form a strip having a plurality of exterior surfaces;

forming a passageway extending through an exterior surface of the strip into the detection chamber so as to form a detection chamber vent;

mounting a layer over the passageway opening to the exterior surface of the strip so as to close the detection chamber vent; and

placing an immobilized binding site and probe conjugate within the reaction chamber.

46. A method of manufacture of the biosensor of claim 28, the method comprising the steps of:

providing a first layer and a second layer;

mounting at least one section of the first layer on a first surface of the second layer to form a filling chamber part, a reaction chamber part, and a detection chamber part;

providing a third layer;

mounting a first side of the third layer to a second side of the first layer, the third layer extending over the filling chamber part, the reaction chamber part and the detection chamber part;

forming a passageway extending through an exterior surface of the strip into the detection chamber so as to form a detection chamber vent;

mounting a layer over the passageway opening to the exterior surface of the strip so as to close the detection chamber vent; and

placing an immobilized binding site and probe conjugate within the reaction chamber.

47. A method of manufacture of the biosensor claim 28, the method comprising the steps of:

providing a first layer;

providing a second layer;

mounting a first side of the second layer to a first side of the first layer;

providing a third layer;

mounting a first side of a third layer to a second side of the first layer;

removing a section of the first layer to form a filling chamber part;

removing a section of the first layer to form a reaction chamber part;

removing a section of the first layer to form a detection chamber part, whereby the reaction chamber and the detection chamber part are in fluid communication with the filling chamber part;

forming a passageway extending through an exterior surface of the strip into the detection chamber so as to form a detection chamber vent;

mounting a layer over the passageway opening to the exterior surface of the strip so as to close the detection chamber vent; and

placing an immobilized binding site and probe conjugate within the reaction chamber.