Process for immunochromatography with colloidal particles

• US 5,120,643 A
• Filed: 07/13/1987
• Issued: 06/09/1992
• Est. Priority Date: 07/13/1987
• Status: Expired due to Term

• Current Assignee: Inverness Medical Switzerland GmbH (Abbott Laboratories)
• Original Assignee: Abbott Laboratories
• Inventors: Billing, Patricia, Gordon, Julian, Ching, Shanfun
• Primary Examiner(s): Saunders, David
• Application Number:
  US07/072,459
• Time in Patent Office: 1,793 Days
• Field of Search: 436/518, 436/528, 436/530, 436/544, 436/801, 436/807, 436/808, 436/824, 436/826, 436/161, 436/162, 436/514, 436/524, 436/525, 436/501, 422/56, 422/59, 422/69, 422/70, 210/656, 210/679, 210/198.2, 210/198.3, 435/7, 435/805, 435/7.93, 435/7.94, 435/7.92, 435/967, 435/970
• US Class Current: 435/7.92
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 Analytical elements
• 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 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• 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
• CPC Class Codes: Y10S 436/801 Electron dense compounds, e.g. ferritin
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives

Abstract

The present invention relates to improved specific binding assay methods, kits and devices utilizing chromatographically mobile specific binding reagents labelled with colloidal particles. Specific binding reagents labelled with colloidal particles such as gold and selenium may be subjected to rapid chromatographic solvent transport on chromatographic media by means of selected solvents and chromatographic transport facilitating agents. Further, impregnation of solid substrate materials with labile protein materials including colloidal particle and enzyme labelled reagents in the presence of meta-soluble proteins provides for the rapid resolubilization of such materials which have been dried onto such substrate materials.

Claims

1. A method for determining the presence or amount of an analyte in a test sample, comprising the steps of:

a) contacting a chromatographic medium with the test sample, said medium comprising at least two reaction sites,

a first reaction site comprising a dried solution of a labelled specific binding reagent in the presence of a meta-soluble protein, and

a second reaction site comprising an immobilized specific binding reagent capable of immobilizing said labelled reagent in relation to the presence or amount of the analyte in the test sample;

b) solubilizing said labelled reagent and transporting at least a portion of said labelled reagent to said second reaction site, and binding said labelled reagent to said second reaction site in relation to the presence or amount of the analyte in the test sample; and

c) detecting said labelled reagent at said second site to determine the presence or amount of the analyte in the test sample.

2. The method according to claim 1 wherein said immobilized specific binding reagent and said labelled reagent specifically bind the analyte.

3. The method according to claim 1 wherein said labelled reagent and the analyte compete in binding to said immobilized specific binding reagent.

4. The method according to claim 1, wherein said meta-soluble protein is selected from the group consisting of casein, zein and meta-soluble non-albumin egg white protein.

5. The method according to claim 4, wherein said meta-soluble protein is casein.

6. The method according to claim 1, wherein said labelled specific binding reagent is a colloidal particle labelled specific binding reagent.

7. The method according to claim 6, wherein said colloidal particles have diameters varying from about 1 nm to about 200 nm.

8. The method according to claim 7, wherein said colloidal particles have diameters ranging from about 40 nm to about 80 nm.

9. The method according to claim 6, wherein said colloidal particles are colloidal metal particles.

10. The method according to claim 9, wherein said colloidal particles are colloidal gold particles.

11. The method according to claim 6, wherein said colloidal particles are colloidal non-metal particles.

12. The method according to claim 11, wherein said colloidal particles are colloidal selenium particles.

13. The method according to claim 1, wherein said labelled specific binding reagent is an enzyme labelled specific binding reagent.

14. The method according to claim 1, wherein said chromatographic medium has a pore size from about 0.05 to about 20 microns.

15. The method according to claim 14, wherein said chromatographic medium comprises nitrocellulose having a pore size of about 5.0 microns.

16. A test device for determining the presence or amount of an analyte in a test sample, comprising:

a chromatographic medium having capillarity and the capacity for transport of non-immobilized reagents and reactive sample components, and comprising at least two reaction sites,

wherein a first reaction site comprises a dried solution of a labelled specific binding reagent in the presence of a meta-soluble protein, said labelled specific binding reagent being solubilized and transported through said chromatographic medium upon contact with a chromatographic solvent or a test sample, and

wherein a second reaction site comprises an immobilized specific binding reagent capable of immobilizing said labelled reagent in relation to the presence or amount of the analyte in the test sample.

17. The test device according to claim 16, wherein said immobilized specific binding reagent and said labelled reagent specifically bind the analyte.

18. The test device according to claim 16, wherein said labelled reagent and the analyte compete in binding to said immobilized specific binding reagent.

19. The test device according to claim 16, wherein said meta-soluble protein is selected from the group consisting of casein, zein and meta-soluble non-albumin egg white protein.

20. The test device according to claim 19, wherein said meta-soluble protein is casein.

21. The test device according to claim 16, wherein said labeled specific binding reagent is a colloidal particle labeled specific binding reagent.

22. The test device according to claim 21, wherein said colloidal particles have diameters ranging from about 1 nm to about 200 nm.

23. The test device according to claim 21, wherein said colloidal particles have diameters ranging from about 40 nm to about 80 nm.

24. The test device according to claim 21, wherein said colloidal particles are colloidal metal particles.

25. The test device according to claim 24, wherein said colloidal particles are colloidal gold particles.

26. The test device according to claim 21, wherein said colloidal particles are colloidal non-metal particles.

27. The test device according to claim 26, wherein said colloidal particles are colloidal selenium particles.

28. The test device according to claim 16, wherein said labeled specific binding reagent is an enzyme labeled specific binding reagent.

29. The device according to claim 16, wherein said chromatographic medium has a pore size from about 0.05 to about 20 microns.

30. The device according to claim 29, wherein said chromatographic medium comprises nitrocellulose having a pore size of about 5.0 microns.

31. A kit for use in specific binding assays for determining the presence or amount of an analyte in a test sample, comprising:

a) a chromatographic medium having a capillarity and the capacity for transport of non-immobilized reagents and reactive sample components, and comprising at least two reaction sites,

wherein a first reaction site comprises a labelled specific binding reagent, in the presence of a meta-soluble protein, wherein said labelled specific binding reagent is capable of being solubilized and transported through said chromatographic medium upon contact with a chromatographic solvent, and

wherein a second reaction site comprises an immobilized specific binding reagent capable of immobilizing said labelled reagent in relation to the presence or amount of the analyte in the test sample; and

b) a chromatographic solvent.