Solid phase chromatographic immunoassay

• US 5,591,645 A
• Filed: 04/20/1993
• Issued: 01/07/1997
• Est. Priority Date: 03/27/1987
• Status: Expired

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Rosenstein, Robert W.
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/049,247
• Time in Patent Office: 1,358 Days
• Field of Search: 435/7.92-7.95, 435/970, 435/310, 435/7.1, 435/805, 436/501, 436/514, 436/515, 436/518, 436/523, 436/524, 436/541, 436/810, 436/829, 436/528, 422/56, 422/58, 422/60, 422/57
• US Class Current: 436/514
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• 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 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

A chromatographic test strip comprising a solid support having at least a first portion and a second portion with said portions being in the same plane so as to permit capillary flow communication with each other. The sample is added to the first portion. The first portion also may comprise a tracer portion having a tracer movably supported therein. The tracer consists of a visible particulate marker. In the second portion, a binder is immobilized. The test strip is useful in a variety of immunoassays.

Claims

1. A test strip for determining the presence of an analyte in a liquid sample comprising a solid support, said solid support comprising at least a first portion and a second portion, said portions being in the same plane so as to permit capillary flow communication with each other;

said first portion being the site for application of the liquid sample and further comprising a tracer site, said tracer site consisting of a tracer movably supported therein wherein said tracer comprises a ligand, which specifically binds to the analyte, conjugated to a visible particulate marker; and

said second portion being the site for visually determining the presence of the visible particulate marker, said second portion consisting of a binder immobilized therein which specifically binds to the analyte.

2. The test strip of claim 1 wherein the solid support comprises nitrocellulose.

3. The test strip of claim 1 wherein the visible particulate marker is selected from the group consisting of colloidal metals, colored liposomes, colored polymeric beads and polymerized dye molecules.

4. The test strip of claim 3 wherein the visible particulate marker is a colored liposome.

5. The test strip of claim 3 wherein the visible particulate marker is a colored polymeric bead.

6. The test strip of claim 1 wherein the analyte is an antigen and the ligand and the binder are antibodies thereto.

7. The test strip of claim 1 wherein the ligand and the binder are antigens or analogs thereof and the analyte is an antibody thereto.

8. The test strip of claim 1 wherein the first portion and the tracer portion are spatially separate from each other with the first portion being upstream of the tracer portion.

9. A no-wash, one-step method for determining the presence of an analyte in a liquid sample consisting of the steps of:

a) adding a liquid sample to the first portion of the test strip of claim 1;

b) allowing sufficient time for the liquid sample to flow to the second portion of the test strip; and

c) determining the presence of the analyte in the liquid sample by visual inspection of the second portion for the visible particulate marker wherein the presence of the analyte is indicated by the presence of the visible particulate marker.

10. The method of claim 9 wherein the liquid sample is added to the test strip by immersing the first portion into the liquid sample.

11. A test strip for determining the amount of an analyte in a liquid sample comprising a solid support, said solid support comprising at least a first portion and a second portion, said portions being in the same plane so as to permit capillary flow communication with each other;

said first portion being the site for application of the liquid sample and further comprising a tracer site, said tracer site consisting of a tracer movably supported therein wherein said tracer comprises a ligand, which is the analyte or an analog thereof, conjugated to a visible particulate marker; and

said second portion being the site for visually determining the amount of the visible particulate marker, said second portion consisting of a binder immobilized therein which specifically binds to the ligand.

12. The test strip of claim 11 which consists of a third portion in the same plane as the first and the second portions, all of said portions being in capillary flow communication with each other, and said third portion being an additional site for visually determining the amount of visible particulate marker bound therein.

13. The test strip of claim 11 wherein the visible particulate marker is selected from the group consisting of colloidal metals, colored liposomes, colored polymeric beads and polymerized dye molecules.

14. The test strip of claim 13 wherein the visible particulate marker is a colored liposome.

15. The test strip of claim 13 wherein the visible particulate marker is a colored polymeric bead.

16. The test strip of claim 11 wherein the analyte is an antigen.

17. The test strip of claim 11 wherein the analyte is an antibody.

18. The test strip of claim 11 wherein the first portion and the tracer portion are spatially separate from each other with the first portion being upstream of the tracer portion.

19. A no-wash, one-step method for determining the presence of an analyte in a liquid sample consisting of the steps of:

a) adding a liquid sample to the first portion of the test strip of claim 11;

b) allowing sufficient time for the liquid sample to flow to the second portion of the test strip; and

c) determining the presence of the analyte in the liquid sample by visual inspection of the second portion for the visible particulate marker wherein the presence of the analyte is indicated by the absence of the visible particulate marker.

20. A no-wash, one-step method for determining the amount of an analyte in a liquid sample consisting of the steps of:

a) adding a liquid sample to the first portion of the test strip of claim 12;

b) allowing sufficient time for the liquid sample to flow to the second portion and the third portions of the test strip; and

c) determining the amount of the analyte present in the liquid sample by visual inspection of the second portion and the third portion for the amount of the visible particulate marker bound in each portion wherein the presence of the analyte is indicated by the absence of the visible particulate marker.

21. The method of claim 19 wherein the liquid sample is added to the test strip by immersing the first portion into the liquid sample.

22. The method of claim 20 wherein the liquid sample is added to the test strip by immersing the first portion into the liquid sample.

23. The test strip of claim 11 wherein the solid support comprises nitrocellulose.