Membrane-based lateral flow assay devices that utilize phosphorescent detection

• US 8,557,604 B2
• Filed: 12/14/2007
• Issued: 10/15/2013
• Est. Priority Date: 11/21/2003
• Status: Active Grant

• Current Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Original Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Inventors: Song, Xuedong
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US11/956,397
• Publication Number:
  US 20080090253A1
• Time in Patent Office: 2,132 Days
• Field of Search: 436/501, 436/518, 436/164, 436/172, 436/543, 436/544, 436/546, 435/546, 435/4, 435/7.1, 435/287.1, 435/287.2, 435/288.7, 422/68.1, 422/82.05, 422/82.08, 422/82.09
• US Class Current: 436/514
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• 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: G01N 33/587 Nanoparticles
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

A lateral flow, membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample is provided. The device utilizes phosphorescence to detect the signals generated by excited phosphorescent labels. The labels may have a long emission lifetime so that background interference from many sources, such as scattered light and autofluorescence, is practically eliminated during detection. In addition, the phosphorescent labels may be encapsulated within particles to shield the labels from quenchers, such as oxygen or water, which might disrupt the phosphorescent signal.

Claims

1. A lateral flow assay device for detecting the presence or quantity of an analyte residing in a test sample, the assay device comprising a porous membrane in fluid communication with detection probes, the detection probes comprising a phosphorescent metal complex encapsulated within a barrier matrix comprising polymethylmethacrylate, the detection probes being conjugated with a specific binding member configured to bind with the analyte and form an analyte complex, wherein the porous membrane defines:

a conjugate pad, the conjugate pad including the detection probes,

a detection zone positioned downstream of the conjugate pad within which a capture reagent is immobilized that is configured to bind to the analyte complex to generate a detection signal, and

a calibration zone positioned downstream of the detection zone within which a capture reagent is immobilized, the capture reagent being capable of binding to uncaptured detection probes,

wherein the analyte is detected by subjecting the detection zone and calibration zone to pulses of light to generate the detection signal and the calibration signal, respectively, and after a certain period of time has elapsed following a pulse of light, the intensity of the detection signal and the calibration signal are measured, the amount of the analyte in the test sample is proportional to a ratio of the intensity of the detection signal to the intensity of the calibration signal.

2. The device as in claim 1, wherein the metal complex comprises a metal selected from the group consisting of ruthenium, osmium, rhenium, iridium, rhodium, platinum, indium, palladium, molybdenum, technetium, copper, iron, chromium, tungsten, zinc, and combinations thereof.

3. The device as in claim 1, wherein the metal complex comprises a ligand selected from the group consisting of pyridine, pyrazine, isonicotinamide, imidazole, bipyridine, terpyridine, phenanthroline, dipyridophenazine, porphyrin, porphine, derivatives thereof, and combinations thereof.

4. The device as in claim 1, wherein the metal complex comprises a porphine complex.

5. The device as in claim 1, wherein the metal complex is selected from the group consisting of platinum (II) tetra-meso-fluorophenylporphine and palladium (II) tetra-meso-fluorophenylporphine.

6. The device as in claim 1, wherein the metal complex comprises platinum (II) tetra-meso-fluorophenylporphine.

7. The device as in claim 1, wherein the metal complex comprises palladium (II) tetra-meso-fluorophenylporphine.

8. The device as in claim 1, wherein the barrier matrix is capable of shielding the encapsulated phosphorescent label from phosphorescence quenchers.

9. The device as in claim 1, wherein the specific binding member is selected from the group consisting of antigens, haptens, aptamers, primary or secondary antibodies, biotin, and combinations thereof.

10. The device as in claim 1, wherein the detection probes are in the form of a particle.

11. The device as in claim 10, wherein the particle is a nanoparticle.

12. The device as in claim 11, wherein the nanoparticle has a diameter of from about 1 nanometer to about 10 microns.

13. The device as in claim 1, wherein the phosphorescent metal complex comprises a phosphorescent label that has a phosphorescent lifetime of greater than about 1 microsecond.

14. The device as in claim 1, wherein the phosphorescent metal complex comprises a phosphorescent label that has a Stokes shift of greater than about 100 nm.

15. The device as in claim 1, wherein the polyelectrolyte capture reagent is a non-biological reagent.

16. The device as in claim 1, further comprising a sampling pad located upstream of the conjugate pad.

17. The device as in claim 1, wherein the capture reagent in the calibration zone is a polyelectrolyte capture reagent.

18. The device of claim 1, wherein the detection zone and the calibration zone are simultaneously subjected to one or more pulses of light.

19. The device of claim 1, wherein the intensity of the detection signal and the intensity of the calibration signal are measured simultaneously.

20. The device of claim 1, wherein the intensity of the detection signal is measured after about 1 to about 100 microseconds.