Analytical test apparatus with on board negative and positive control

• US 5,356,782 A
• Filed: 09/03/1992
• Issued: 10/18/1994
• Est. Priority Date: 09/03/1992
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics Operations Incorporated (Roche Holding AG)
• Original Assignee: Boehringer Mannheim Corporation (Roche Holding AG)
• Inventors: Heald, Brian A., Webster, David D., Ledden, David J., Moorman, David R.
• Primary Examiner(s): Scheiner, Toni R.
• Application Number:
  US07/940,059
• Time in Patent Office: 775 Days
• Field of Search: 422/56, 422/57, 422/68.1, 422/69, 422/61, 435/5, 435/6, 435/7, 435/4, 435/7.1, 435/7.5, 435/7.9, 435/7.92, 435/805, 435/967, 435/970, 435/975, 436/4, 436/165, 436/166, 436/168, 436/169, 436/820, 436/501, 436/512, 436/518, 436/523, 436/536
• US Class Current: 435/7.9
• 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 435/805 Test papers
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/975 Kit

Abstract

The invention is an apparatus useful in carrying out an analytical assay. The apparatus has a positive and negative control, as well as a site for determining the presence, amount, or lack of an analyte in a sample.

Claims

1. An apparatus for determining an analyte of interest in a liquid sample comprising an absorptive material and having a plurality of zones in the direction of fluid flow, wherein:

(i) a first negative control zone of said absorptive material contains an immobilized reactant which does not bind to said analyte of interest;

(ii) a second analytical zone of said absorptive material contains an immobilized reactant which specifically binds to said analyte of interest; and

(iii) a third positive control zone of said absorptive material contains both (a) an immobilized reactant which specifically binds to said analyte of interest, and (b) a portion of the analyte of interest in solubilizible form, wherein said portion of the analyte is solubilized when contacted with the liquid sample and reacts with the immobilized reactant of said third zone.

2. The apparatus of claim 1, wherein the immobilized reactant of said second zone is an immobilized antibody or an antibody fragment.

3. The apparatus of claim 1, wherein said immobilized reactant is bound to a carrier which is immobilized in said second zone.

4. The apparatus of claim 3, wherein said carrier comprises a material selected from the group consisting of latex, glass, sepharose, agarose, polystyrene, mica and diatomaceous earth.

5. The apparatus of claim 3, wherein said carrier is a bead.

6. The apparatus of claim 5, wherein said carrier has a diameter of from 1.0 nm to 20 μ

m.

7. The apparatus of claim 6, wherein said carrier has a diameter of from 0.3 to 1.0 μ

m.

8. The apparatus of claim 7, wherein said carrier has a diameter of from 0.4 to 0.5 μ

m.

9. The apparatus of claim 1, wherein the immobilized reactant of said second zone is selected from the group consisting of an antigen and an epitope containing fragment of an antigen.

10. The apparatus of claim 1, wherein said second zone contains at least one substance selected from the group consisting of protein A, protein G, streptavidin, avidin, and a lectin.

11. The apparatus of claim 1, wherein the immobilized reactants of said second zone and said third zone are identical.

12. The apparatus of claim 1, wherein the immobilized reactant of said third zone and the immobilized reactant of said second zone are identical, and the immobilized reactant of said first zone is an inactive form of the immobilized reactants of said second and third zones, wherein said inactive form cannot specifically bind the analyte.

13. The apparatus of claim 1, wherein said immobilized reactant in said second zone is in a position perpendicular to the direction of fluid flow in said absorptive material.

14. The apparatus of claim 1, wherein said immobilized reactant in said third zone is in a position perpendicular to the direction of the fluid flow in said absorptive material.

15. The apparatus of claim 1, further comprising an additional zone positioned downstream of said third zone for absorption of excess fluid.

16. The apparatus of claim 15, wherein said additional zone contains a reagent which changes color in the presence of a fluid.

17. The apparatus of claim 15, wherein said additional zone contains a reagent which changes color when subjected to a pH change.

18. The apparatus of claim 15, wherein said additional zone contains a reagent which inhibits formation of detectable signal.

19. A kit for determining an analyte in a sample comprising the apparatus of claim 1 and a separate portion of a second, labeled receptor which specifically binds to said analyte of interest.

20. The kit of claim 19, wherein said labeled receptor is labeled with a label selected from the group consisting of an enzyme, gold, a dye sol, a colored particle, a fluorescer, a chemiluminescer, and a radiolabel.

21. The kit of claim 19, further comprising a separate portion of a substance which reacts with said labeled receptor to form a detectable signal.

22. The kit of claim 19, further comprising a separate portion of an inert run buffer.

23. The kit of claim 19, wherein said second, labeled receptor is an antibody.

24. The apparatus of claim 1, further comprising a further zone positioned upstream of said first zone, said further zone containing a substance which reacts with a labeled receptor to provide a detectable signal, and an absorptive means positioned on top of said further zone and in fluid contact therewith, wherein said further zone is in fluid contact with said first zone.

25. The apparatus of claim 24, further comprising a fluid flow inhibition means positioned so as to prevent fluid flow from said absorptive means into said first zone and to direct fluid flow from said absorptive means into said further zone.

26. The apparatus of claim 25, further comprising a second fluid flow inhibition means positioned between said further and first zones and extending along all of a region of fluid contact therebetween.

27. The apparatus of claim 24, further comprising a fluid flow regulating means positioned between said further and first zones which directs fluid flow vertically.

28. The apparatus of claim 1, further comprising an inert support on which said first, second and third zones are positioned.

29. The apparatus of claim 1, wherein the immobilized reactant of said second zone is an immobilized antibody fragment.

30. The kit of claim 19 wherein said second, labelled receptor is an antibody fragment.