Membrane-based assay devices

• US 7,285,424 B2
• Filed: 08/27/2002
• Issued: 10/23/2007
• Est. Priority Date: 08/27/2002
• Status: Active Grant

• Current Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Original Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Inventors: Song, Xuedong, Kaylor, Rosann
• Primary Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US10/228,836
• Publication Number:
  US 20040043511A1
• Time in Patent Office: 1,883 Days
• Field of Search: 435/7.92, 435/7.1, 435/7.2, 435/2, 435/5, 435/7.21, 435/7.32, 435/239, 435/803, 435/805, 435/287.2, 435/81, 435/287.9, 435/970, 436/513, 436/512, 436/518, 436/531, 436/526, 436/533, 436/538, 436/806, 436/824, 422/56
• US Class Current: 436/514
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/274 Calibration, base line adjustment, drift correction
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/824 Immunological separation techniques
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

A membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample is provided. The device utilizes a self-calibrated magnetic binding assay format (e.g., sandwich, competitive, etc.) that includes detection probes capable of generating a detection signal (e.g., fluorescent non-magnetic particles) and calibration probes capable of generating a calibration signal (e.g., fluorescent magnetic particles). The amount of the analyte within the test sample is proportional (e.g., directly or inversely) to the intensity of the detection signal calibrated by the intensity of the calibration signal. It has been discovered that the fluidics-based device of the present invention provides an accurate, inexpensive, and readily controllable method of determining the presence of an analyte in a test sample.

Claims

1. A membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample, the device comprising:

detection probes that comprise non-magnetic particles conjugated with a first specific binding member and labeled with a detectable luminescent substance that is configured to emit a detection signal, the first specific binding member being capable of binding with the analyte;

calibration probes that comprise magnetic particles conjugated with a second specific binding member and labeled with a detectable luminescent substance that is configured to emit a calibration signal that is distinguishable from the detection signal, the second specific binding member being capable of binding with the analyte;

a porous membrane in fluid communication with the detection probes and the calibration probes, wherein the porous membrane defines a detection zone; and

a magnetic device positioned adjacent to the detection zone, the magnetic device being configured to immobilize the calibration probes within the detection zone, the magnetic device further being configured to immobilize within the detection zone complexes formed from the detection probes, the analyte, and the calibration probes.

2. The assay device of claim 1, further comprising a conjugate pad in fluid communication with the porous membrane and positioned upstream from the detection zone, the conjugate pad comprising the detection probes and calibration probes.

3. The assay device of claim 1, wherein the porous membrane is in fluid communication with a sampling pad that is positioned upstream from the detection zone.

4. The assay device of claim 1, further comprising a wicking pad in fluid communication with the porous membrane and positioned downstream from the detection zone.

5. The assay device of claim 1, wherein the detectable luminescent substance of both the detection probes and the calibration probes is a fluorescent compound.

6. The assay device of claim 1, wherein the first specific binding member, second specific binding member, or both include an antigen, hapten, aptamer, antibody, or a combination thereof.

7. The assay device of claim 1, wherein the amount of the analyte within the test sample is directly proportional to the intensity of the detection signal produced at the detection zone calibrated by the intensity of the calibration signal produced at the detection zone.

8. The assay device of claim 1, wherein the detectable luminescent substance of both the detection probes and the calibration probes includes a phosphorescent compound.

9. The assay device of claim 1, wherein the detectable luminescent substance of both the detection probes and the calibration probes includes a fluorescent compound.

10. A membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample, the device comprising:

detection probes that comprise non-magnetic particles conjugated with a first binding member and labeled with a detectable luminescent substance that is configured to emit a detection signal, the first specific binding member being capable of binding with the analyte;

calibration probes that comprise magnetic particles labeled with a detectable luminescent substance that is configured to emit a calibration signal that is distinguishable from the detection signal;

non-labeled magnetic particles conjugated with a second binding member, the second binding member also being capable of binding with the analyte;

a porous membrane in fluid communication with the detection probes, the calibration probes, and the second magnetic particles, wherein the porous membrane defines a detection zone; and

a magnetic device positioned adjacent to the detection zone, the magnetic device being configured to immobilize the calibration probes within the detection zone, the magnetic device further being configured to immobilize within the detection zone complexes formed from the detection probes, the analyte, and the non-labeled magnetic particles.

11. The assay device of claim 10, wherein the magnetic particles of the calibration probes are treated with a blocking agent.

12. The assay device of claim 11, wherein the blocking agent includes β

-casein, albumins, polyethylene glycol, polyvinyl alcohol, or combinations thereof.

13. The assay device of claim 10, further comprising a conjugate pad in fluid communication with the porous membrane and positioned upstream from the detection zone, the conjugate pad comprising the detection probes and calibration probes.

14. The assay device of claim 10, wherein the porous membrane is in fluid communication with a sampling pad that is positioned upstream from the detection zone.

15. The assay device of claim 10, further comprising a wicking pad in fluid communication with the porous membrane and positioned downstream from the detection zone.

16. The assay device of claim 10, wherein the detectable substance of both the detection probes and the calibration probes is a fluorescent compound.

17. The assay device of claim 10, wherein the first specific binding member, second specific binding member, or both include an antigen, hapten, aptamer, antibody, or a combination thereof.

18. The assay device of claim 10, wherein the amount of the analyte within the test sample is directly proportional to the intensity of the detection signal produced at the detection zone calibrated by the intensity of the calibration signal produced at the detection zone.

19. The assay device of claim 10, wherein the detectable luminescent substance of both the detection probes and the calibration probes includes a phosphorescent compound.

20. The assay device of claim 10, wherein the detectable luminescent substance of both the detection probes and the calibration probes includes a fluorescent compound.