Membrane-based lateral flow assay devices that utilize phosphorescent detection

• US 8,703,504 B2
• Filed: 09/04/2013
• Issued: 04/22/2014
• Est. Priority Date: 11/21/2003
• Status: Expired due to Term

• 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:
  US14/017,342
• Publication Number:
  US 20140004506A1
• Time in Patent Office: 230 Days
• 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 33/54388 based on lateral flow
• 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 33/582 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 method for detecting an analyte within a test sample, the method comprising:

i) providing a lateral flow assay device that comprises a porous membrane in fluid communication with detection probes, the detection probes comprising a phosphorescent particle, the phosphorescent particle 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 is immobilized a capture reagent 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 to generate a calibration signal;

ii) contacting the lateral flow assay device with the test sample;

iii) subjecting the detection zone to illumination pulses to generate the detection signal;

iv) subjecting the calibration zone to illumination pulses to generate the calibration signal; and

v) thereafter, measuring the intensity of the detection signal and the calibration signal, wherein the amount of the analyte within the test sample is proportional to a ratio of the intensity of the detection signal to the intensity of the calibration signal.

2. A method 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. A method 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. A method as in claim 1, wherein the metal complex comprises a porphine complex.

5. A method 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. A method as in claim 1, wherein the particle comprises a nanoparticle.

7. A method as in claim 6, wherein the nanoparticle has a diameter of from about 1 nanometer to about 10 microns.

8. A method as in claim 1, wherein the barrier matrix shields the encapsulated phosphorescent label from phosphorescence quenchers.

9. A method 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 method as in claim 1, wherein the metal complex comprises platinum(II) tetra-meso-fluorophenylporphine.

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

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

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

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

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

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

17. The method as in claim 1, wherein the detection zone and the calibration zone are simultaneously subjected to one or more pulses of light.

18. The method as in claim 1, wherein the intensity of the detection signal and the intensity of the calibration signal are measured simultaneously.

19. The method as in claim 1, wherein the intensity of the detection signal is measured after about 1 to about 100 microseconds.