Non-instrumented assay with quantitative and qualitative results

• US 5,998,221 A
• Filed: 09/25/1996
• Issued: 12/07/1999
• Est. Priority Date: 09/25/1996
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: McFarland, Edward Charles, Malick, Adrien Paul
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US08/719,223
• Time in Patent Office: 1,168 Days
• Field of Search: 422/56-61, 435/7.1, 435/7.2, 435/7.93, 435/7.94, 435/7.95, 435/287.7, 435/288.6, 435/969, 435/970, 435/973, 436/518, 436/530, 436/514
• US Class Current: 436/514
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• 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 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte

Abstract

The invention is directed to a non-instrumented assay giving quantitative and/or qualitative results. Specifically, the invention is directed to an assaying process wherein a first portion of binder supported on substantially the entire length of a solid is contacted with a sample suspected of containing analyte. The binder is a binder for at least the analyte. The first portion is also contacted with a tracer. The process includes contacting a second binder portion also supported on the solid with tracer. The second portion is juxtaposed to the first portion and has a known amount of analyte thereon. The amount of analyte in the sample is determined by comparing the visibility of tracer in the first and second portions. In a preferred embodiment, a third binder portion supported on the solid juxtaposed to the second portion is contacted with tracer. The third portion has a known amount of analyte thereon. The amount of analyte in the third portion is different from the amount on the second portion. According to another aspect, the invention is also directed to a kit for performing the above-described process.

Claims

1. A process for assaying for an analyte in a sample, said process comprising:

providing a solid support having at least two lanes wherein each lane has a contact zone for applying sample to each lane and further wherein the first lane of the solid support has binder immobilized on the entire length of the first lane of the solid support, said binder being a binder for at least the analyte, and the second lane of the solid support is juxtaposed to the first lane of the solid support, and said second lane has binder immobilized on the entire length of said second lane of the solid support, said binder being a binder for at least the analyte, and a known amount of said analyte is immobilized on said binder which is immobilized on the entire length of the second lane of the solid support;

contacting the contact zone of the first lane with sample and tracer wherein said tracer is comprised of a ligand labeled with a visible label;

contacting the contact zone of the second lane with tracer; and

determining the visibility of tracer bound in the first and second lanes of binder as a measure of analyte in the sample.

2. The process of claim 1 wherein sample and tracer are sequentially contacted with the contact zone of the first lane of the solid support.

3. The process of claim 1 wherein the sample and tracer are simultaneously contacted with the contact zone of the first lane of the solid support.

4. The process of claim 1 wherein each lane is separated by an impermeable wall or membrane.

5. The process of claim 1 wherein the visible label comprises a sac including a visible dye.

6. The process of claim 5 wherein the sac is a liposome.

7. The process of claim 6 wherein the visibility of the bound tracer is determined without lysis of the liposome.

8. The process of claim 1 wherein the solid support is in sheet form.

9. The process of claim 1 wherein the solid support is comprised of nitrocellulose.

10. The process of claim 9 wherein the solid support is a nitrocellulose test card.

11. The process of claim 1 wherein the binder is an antibody.

12. The process of claim 11 wherein the tracer is a labeled form of the analyte.

13. The process of claim 1 1 wherein the tracer is a labeled form of an antibody for the analyte.

14. The process of claim 1 wherein the analyte is digoxin.

15. The process of claim 1 wherein the visible label comprises a colored particle.

16. The process of claim 1 wherein said assay has a third lane with a contact zone, wherein said third lane is juxtaposed to the first or second lane and further comprising contacting the contact zone of said third lane with tracer, said third lane having binder immobilized on the entire length of said third lane and having a known amount of analyte immobilized on said binder which is immobilized on the entire length of the third lane, said amount of analyte in the third lane being different from the amount of analyte on the second lane.

17. The process of claim 16 wherein said assay has one to ten more lanes each having a contact zone.

18. The process of claim 17 wherein each lane is juxtaposed to the other, and each lane has binder immobilized on the entire length of said lane and having a known amount of analyte immobilized on said binder which is immobilized on the entire length of said lane, said amount of analyte being different from any other lane.

19. The process of claim 16 wherein each lane is separated by an impermeable wall or membrane.

20. The process of claim 18 wherein each lane is separated by an impermeable wall or membrane.

21. A kit for assaying for an analyte in a sample comprising:

a solid support having at least two lanes wherein each lane has a contact zone for applying sample to each lane and further wherein the first lane of the solid support has binder immobilized on the entire length of said lane of the solid support, and the second lane of the solid support is juxtaposed to the first lane of the solid support, and said second lane has binder immobilized on the entire length of said second lane of the solid support and a known amount of analyte is immobilized on said binder which is immobilized on the entire length of the second lane of the solid support;

binder, said binder being a binder for at least the analyte;

a tracer comprised of a ligand labeled with a visible label, wherein said tracer together with sample is contacted to the contact zone of the first lane or portion and wherein tracer is contacted to the contact zone of the second lane or portion after sample has been added thereon;

and further, said binder being supported in a concentration whereby bound tracer is visible on the support under assay conditions.

22. The kit of claim 21 wherein each lane is separated by an impermeable wall or membrane.

23. The kit of claim 21 wherein the binder is an antibody.

24. The kit of claim 21 wherein the visible label of the tracer is a liposome, including a visible dye.

25. The kit of claim 21 wherein the analyte is digoxin, the binder is an antibody to digoxin and the ligand of the tracer is an analog of digoxin.

26. The kit of claim 21 further comprising a third lane having binder immobilized on the entire length of said lane on said solid support juxtaposed to either the first or second binder lanes wherein said third lane includes a known amount of analyte immobilized on said binder which is immobilized on the entire length of the third lane, said amount being different than the amount on said second lane.

27. The kit of claim 24 further comprising one to ten more lanes juxtaposed to said third lane having binder immobilized on the entire length of said lane, said further lane or lanes including a known amount of analyte immobilized on said binder which is immobilized on the entire length of said lane, said amount being different than the amounts on said second and third lanes.

28. The kit of claim 26 wherein each lane is separated by an impermeable wall or membrane.

29. The kit of claim 27 wherein each lane is separated by an impermeable wall or membrane.