Membrane-Based Lateral Flow Assay Devices that Utilize Phosphorescent Detection

• US 20080090253A1
• Filed: 12/14/2007
• Published: 04/17/2008
• 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
• Granted Patent: US 8,557,604 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• 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 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 a phosphorescent particle, the phosphorescent particle comprising a phosphorescent label encapsulated within a barrier matrix comprising polymethylmethacrylate, the phosphorescent label capable of emitting a detection signal following excitation of the phosphorescent label, wherein the porous membrane defines a detection zone within which is immobilized a capture reagent;

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

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

iv) thereafter, measuring the intensity of the detection signal, wherein the amount of the analyte within the test sample is proportional to the intensity of the detection 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 phosphorescent particle is conjugated with a specific binding member for an analyte.

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

11. 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, wherein the porous membrane defines a detection zone within which a capture reagent is immobilized that is configured to bind to the detection probes or complexes thereof to generate a detection signal, wherein the amount of the analyte in the test sample is proportional to the intensity of the detection signal.

12. A device as in claim 11, wherein the metal complex comprises platinum(II) tetra-meso-fluorophenylporphine.

13. A device as in claim 11, wherein the metal complex comprises palladium(II) tetra-meso-fluorophenylporphine.

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

15. A device as in claim 11, wherein the barrier matrix is capable of shielding the encapsulated phosphorescent label from phosphorescence quenchers.

16. A device as in claim 11, wherein the specific binding member is selected from the group consisting of antigens, haptens, aptamers, primary or secondary antibodies, biotin, and combinations thereof.