Chromatographic test strip for determining ligands or receptors

• US 4,960,691 A
• Filed: 09/29/1986
• Issued: 10/02/1990
• Est. Priority Date: 09/29/1986
• Status: Expired due to Term

• Current Assignee: Abbott Laboratories North Chicago IL
• Original Assignee: Abbott Laboratories
• Inventors: Ching, Shanfun, Gordon, Julian, McMahon, Michael E.
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): Saunders, David A.
• Application Number:
  US06/912,878
• Time in Patent Office: 1,464 Days
• Field of Search: 435/67, 435/805, 436/501, 436/518, 436/530, 436/807, 436/810, 436/162, 436/517, 422/56, 422/58, 422/69, 422/61, 422/70
• US Class Current: 435/6.12
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: G01N 30/38 Flow patterns
• CPC Class Codes: G01N 30/6065 with varying cross section
• CPC Class Codes: G01N 30/92 Construction of the plate
• 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 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

A test strip for analysis of analytes such as antigens, antibodies or polynucleotides employs a chromatographic medium and a solvent capable of transporting reagents and/or sample. Reagents are selected and disposed on the medium such that a labeled (first) reagent arrives at the detection (third) zone only after analyte is immobilized there and non-reactive sample componets have been transported beyond the detection zone. This sequential arrival is accomplished by the relative mobility of the reagent or sample; or by the site relationship of the zones. Preferably, the site relationship of the sample (second) zone and the label (first) zone is such that a plurality of pathways guide the sample and labeled reagent and any subsequent reagents to the detection zone in the recited order. Single and multiple pathway devices are disclosed.

Claims

1. A test strip for analysis of an analyte in a sample by means of a sequential series of reactions, said strip comprising:

a length of chromatographic material having capillarity and the capacity for chromatographic solvent transport of non-immobilized reagents and reactive sample components by means of a selected chromatographic solvent, said strip including,

a first end at which chromatographic solvent transport begins;

a second end, at which chromatographic solvent transport ends;

a plurality of chromatographic solvent transport pathways;

a plurality of zones positioned between said first and second ends, said zones including,

a first zone impregnated with a first reagent which is mobile in said solvent and capable of a specific binding reaction with said analyte,

a second zone for receiving said sample, and

a third zone, downstream of said second zone, impregnated with a second reagent which is immobilized against solvent transport and is capable of a specific binding reaction with the analyte so as to immobilize the analyte in said third zone, said first and second zones being spaced sufficiently from said first end to permit contact of said first end but not said first and second zones with said chromatographic solvent, and

means for detecting said first reagent at said third zone as a measure of the analyte,

whereby, after said first end is dipped into said chromatographic solvent, the relative mobility of said sample components and said first reagent is controlled by said plurality of chromatographic solvent transport pathways such that the analyte is transported to and immobilized against solvent transport at the third zone prior to the first reagent reaching the third zone, and whereby interfering sample components and non-analyte components of the sample which are reactive with said first reagent are cleared from said third zone by chromatographic solvent transport prior to the first reagent reaching said third zone.

2. The test strip according to claim 1 wherein said means for detecting said first reagent at said third zone is a label on said first reagent.

3. The test strip according to claim 2 wherein said label is selected from the group consisting of radiolabels, chromophores, fluorophores and enzyme labels.

4. The test strip according to claim 1 wherein the chromatographic solvent transport pathways of said sample and of said first reagent are partially non-coincident.

5. The test strip according to claim 3 wherein said label is an enzyme label and a third reagent reactive with said label is disposed at a fourth zone and has a relative mobility in said chromatographic solvent such that the third reagent is transported to said third zone by said solvent after said first reagent is transported to said third zone.

6. The test strip according to claim 5 wherein said third reagent is an indicator substance.

7. The test strip according to claim 5 wherein the chromatographic solvent transport pathways of said first reagent and said third reagent are partially non-coincident.

8. The test strip according to claim 1 wherein said chromatographic solvent transport pathways are separated by means of solvent impermeable barriers.

9. The test strips according to claim 8 wherein said solvent impermeable barriers are formed by laser etching.

10. The test strips according to claim 1 wherein said chromatographic material is thin layer chromatographic material.

11. The test strips according to claim 10 wherein said thin layer chromatographic material is nitrocellulose.

12. The test strips according to claim 1 wherein said analyte is an antibody.

13. The test strips according to claim 1 wherein said analyte is an antigen.

14. The test strips according to claim 1 wherein said analyte is a polynucleotide.

15. A method for analysis of analyte in a sample by means of a sequential series of reactions, said method employing:

a strip comprising a length of chromatographic material having capillarity and the capacity for chromatographic solvent transport of non-immobilized reagents and reactive sample components by means of a selected chromatographic solvent, said strip including,

a first end at which chromatographic solvent transport begins;

a second end, at which chromatographic solvent transport ends;

a plurality of chromatographic solvent transport pathways;

a plurality of zones positioned between said first and second ends, said zones including,

a first zone impregnated with a first reagent which is mobile in said solvent and capable of a specific binding reaction with said analyte,

a second zone for receiving said sample, and

a third zone, downstream of said second zone, impregnated with a second reagent which is immobilized against solvent transport and is capable of a specific binding reaction with the analyte so as to immobilize the analyte in said third zone, said first and second zones being spaced sufficiently from said first end to permit contact of said first end but not said first and second zones with said chromatographic solvent, and

means for directing said first reagent at said third zone,

said method comprising;

(1) disposing said sample in said second zone;

(2) dipping said first end into chromatographic solvent for a time sufficient to chromatographically transport said analyte and said first reagent to said third zone, wherein the relative mobility of said sample components and said first reagent is controlled by said plurality of chromatographic solvent transport pathways such that the analyte is transported to and immobilized against solvent transport at the third zone prior to the first reagent reaching the third zone, and whereby interfering sample components and non-analyte components of the sample which are reactive with said first reagent are cleared from said third zone by the chromatographic solvent transport prior to the first reagent reaching said third zone; and

(3) detecting the presence of said first reagent in said third zone as a measures of the analyte.

16. A method according to claim 15 wherein said means for detecting said first reagent at said third zone is a label on said first reagent.

17. A method according to claim 15 wherein said means for detecting said first reagent at said third zone is a third reagent reactive with said first reagent.

18. A method according to claim 17 wherein the third reagent is an indicator substance.

19. A method according to claim 17 wherein said third reagent is disposed at a fourth zone and is chromatographically transported to said third zone after said first reagent is transported to said third zone.

20. A method according to claim 19 wherein said first reagent and said third reagent are transported along pathways of travel which are partially noncoincident.

21. A method according to claim 19 wherein said sample and said third reagent are transported along pathways of travel which are partially non-coincident.

22. A method according to claim 15 wherein said chromatographic solvent pathways are partially noncoincident as a result of solvent impermeable barriers.

23. A method according to claim 15 wherein said chromatographic material is thin layer chromatographic material.

24. A method according to claim 23 wherein said thin layer chromatographic material is nitrocellulose.

25. A method according to claim 24 wherein said nitrocellulose is characterized by an average pore size ranging from about 0.1 micron to about 1 micron.

26. A method according to claim 15 wherein said analyte is an antibody.

27. A method according to claim 15 wherein said analyte is an antigen.

28. A method according to claim 15 wherein said analyte is a polynucleotide.

29. A method for analysis of analyte in a sample by means of a sequential series of reactions, said method employing:

a strip comprising a length of chromatographic material having capillarity and the capacity for chromatographic solvent transport of non-immobilized reagents and reactive sample components by means of a selected chromatographic solvent, said strip including,

a first end at which chromatographic solvent transport begins;

a second end, at which chromatographic solvent transport ends;

a plurality of zones positioned between said first and second ends, said zones including,

a first zone impregnated with a first reagent which is mobile in said solvent and capable of a specific binding reaction with an analyte,

a second zone for receiving said sample, and

a third zone, coincident with said second zone, impregnated with a second reagent which is immobilized against solvent transport and is capable of a specific binding reaction with the analyte so as to immobilize the analyte in said third zone, said first and second zones being spaced sufficiently from said first end to permit contact of said first end but not said first and second zones with said chromatographic solvent, and

means for detecting said first reagent at said third zone as a measure of the analyte,

said method comprising;

(1) disposing said sample in said second zone;

(2) dipping said first end into said chromatographic solvent for a time sufficient to chromatographically transport said first reagent to said third zone, whereby after said first end is dipped into said chromatographic solvent, the relative mobility of said sample components and said first reagent is such that the analyte is immobilized against solvent transport at the third zone prior to the first reagent reaching the third zone, and whereby interfering sample components and non-analyte components of the sample which are reactive with said first reagent are cleared from said third zone by chromatographic solvent transport prior to the first reagent reaching said third zone, and (3) detecting the presence of said first reagent in said third zone as a measure of the analyte.

30. A method according to claim 29 wherein said means for detecting said first reagent at said third zone comprises a third reagent reactive with said first reagent.

31. A method according to claim 30 wherein said third reagent is disposed at a fourth zone and is chromatographically transported to said third zone after said first reagent is transported to said third zone.

32. A method according to claim 31 wherein said first reagent and said third reagent are transported along pathways of travel which are partially non-coincident.

33. A method according to claim 29 wherein said chromatographic material comprises thin layer nitrocellulose.