Method for immunochromatographic analysis

• US 5,164,294 A
• Filed: 07/07/1989
• Issued: 11/17/1992
• Est. Priority Date: 05/17/1988
• Status: Expired due to Term

• Current Assignee: DADE Behring Beteiligungs GmbH (Siemens AG)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Ullman, Edwin F., Shanafelt, Armen B., Ghazarossian, Vartan, Skold, Carl N.
• Primary Examiner(s): Saunders, David
• Application Number:
  US07/376,723
• Time in Patent Office: 1,229 Days
• Field of Search: 435/4, 435/7.8, 435/7.92, 435/7.93, 435/805, 435/970, 435/7.5, 436/501, 436/514, 436/518, 436/541, 436/810, 436/161, 436/162, 436/169, 436/170, 436/73, 422/56
• US Class Current: 435/7.5
• CPC Class Codes: G01N 33/54386 Analytical elements
• 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 436/81 Tube, bottle, or dipstick

Abstract

Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present. The distance the component has migrated into the second zone or the difference in the distances the medium and the component have migrated into the second zone is determined, the distance or the difference being related to the amount of the component in the liquid medium or the amount of the reagent.

Claims

1. A method for determining the presence or amount of an analyte, which comprises:

causing a reagent in a first liquid medium to become bound to a first bibulous member zone in relation to the presence or amount of an analyte by contacting said first bibulous member zone with said medium under conditions wherein said first medium flows through or traverses said first bibulous member zone by capillary action;

causing a component in a second liquid medium to be absorbed by a second bibulous member zone in relation to the presence of said analyte in said first medium by contacting a portion of said first bibulous member zone with said second medium, under conditions wherein said second medium traverses said first bibulous member zone and at least a portion of said second bibulous member zone by capillary action; and

determining the distance said component traverses said second bibulous member zone to determine the amount of said analyte, or determining the presence of said component on at least a portion of said second bibulous member to determine the presence of said analyte, said first bibulous member zone being caused to come into liquid receiving relationship with said second bibulous member zone after said reagent is caused to become bound to said first bibulous member zone.

2. The method of claim 1 wherein said first bibulous member zone and said second bibulous member zone are brought into liquid receiving relationship prior to contacting said first bibulous member zone with said second medium.

3. The method of claim 1 wherein said second bibulous member zone is an immunochromatographic member

4. The method of claim 1 wherein a binding partner capable of binding said reagent is non-diffusively bound to at least a portion of said first bibulous member zone.

5. The method of claim 1 wherein said reagent and said analyte bind together to form a complex and said complex becomes bound to said first bibulous member zone.

6. The method of claim 1 wherein a binding partner for said component is non-diffusively bound to said second bibulous member zone.

7. The method of claim 1 where said analyte is an sbp member.

8. A method for conducting an assay for an analyte, said method comprising:

providing in combination in a first liquid medium (1) a first bibulous member, at least a portion thereof having a first reagent non-diffusively bound thereto and (2) a sample suspected of containing an analyte with the proviso that, when said analyte is not capable of binding to said first reagent, a second reagent capable of binding to said analyte and to said first reagent is present in said first liquid medium or on said first bibulous member, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first liquid medium traverses said first bibulous member and at least a portion of said absorbent member by capillary action;

bringing said first bibulous member into liquid receiving relationship with a second bibulous member;

providing in combination in a second liquid medium a portion of said first bibulous member and a component, said component being capable of binding to said first reagent in relation to the presence of said analyte in the first liquid medium, said second bibulous member having non-diffusively and uniformly bound to at least a portion thereof a specific binding partner for said component, under conditions wherein said second liquid medium transverses at least a portion of said first bibulous member and at least a portion of said second bibulous member by capillary action; and

determining the presence of said component on said second bibulous member, the presence thereof being related to the presence of said analyte in said sample, or determining the distance of migration of said component on said second bibulous member, the distance being related to the amount of said analyte in said sample.

9. The method of claim 8 wherein said liquid receiving relationship between said first bibulous member and said absorbent member is terminated prior to contacting said second bibulous member with said second medium.

10. The method of claim 8 wherein said second bibulous member is an immunochromatograph and said component includes an enzyme and said amount of said component bound to said immunochromatographic member is determined by the location on said immunochromatographic member of a border.

11. The method of claim 10 wherein said immunochromatographic member is a strip of bibulous material.

12. The method of claim 10 wherein said immunochromatographic member is contacted with an enzyme substrate solution to determine said border.

13. The method of claim 8 wherein said analyte is a drug.

14. The method of claim 8 wherein said analyte is digoxin.

15. The method of claim 8 wherein said second bibulous member is a paper strip.

16. The method of claim 8 wherein said second reagent comprises (1) a first sbp member and (2) a second sbp member complementary to said first reagent.

17. The method of claim 8 wherein said first reagent comprises an antibody for said analyte and a molecule selected from the group consisting of fluorescein, LSD, biotin, morphine, benzoylecgonine, avidin, intrinsic factor, and folate binding protein.

18. The method of claim 8 wherein said second reagent comprises an antibody for said analyte conjugated to an organic molecule of molecular weight less than 1500.

19. The method of claim 18 wherein said first reagent is an antibody for said organic molecule.

20. The method of claim 8 wherein said absorbent member is a strip of bibulous material.

21. The method of claim 8 wherein said absorbent material has a liquid holding capacity to absorb all of the liquid in said first liquid medium.

22. The method of claim 8 wherein said component is comprised of said analyte bound to a label.

23. The method of claim 22 wherein said label is an enzyme.

24. The method of claim 8 wherein said specific binding partner for said component is an antibody.

25. The method of claim 8 wherein said first reagent is an analyte analog.

26. The method of claim 8 wherein said second reagent is an antibody for said analyte.

27. The method of claim 8 wherein said component is comprised of an antibody for said first reagent bound to a label.

28. The method of claim 27 wherein said label is an enzyme.

29. The method of claim 8 wherein said specific binding partner is a receptor for an immunoglobulin.

30. The method of claim 8 wherein said first reagent is antibody for said analyte.

31. A method for conducting an assay, said method comprising:

combining a solution of a drug with a first bibulous member wherein an antibody for said drug is or becomes nondiffusively bound to said bibulous member,

combining a liquid medium containing an enzyme labeled drug analog with said first bibulous member wherein said component is interreactive with said reagent, said first bibulous member being in liquid receiving relationship with or being a portion of a second bibulous member of a composition different from said first bibulous member and incapable of specifically binding said enzyme labeled drug analog, under conditions wherein said liquid medium transverses said first bibulous member and at least a portion of said second bibulous member by capillary action, and

determining a border on said second bibulous member, said border being defined by a signal produced by the enzyme label and the position of said border being related to the amount of drug in said solution or the amount of antibody for said drug on said first bibulous member.

32. The method of claim 31 wherein said first bibulous member is in liquid receiving relationship with said second bibulous member.

33. A method for conducting an assay, said method comprising:

providing in combination in a first liquid medium (1) a first bibulous member and (2) an analyte, wherein a reagent for said analyte is present in said medium or on said first bibulous member, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium flows through said first bibulous member and at least a portion of said absorbent member by capillary action and said analyte becomes non-diffusively bound to said bibulous member;

terminating the liquid receiving relationship between said bibulous member and said absorbent member;

providing in combination in a second liquid medium a portion of said first bibulous member and a component interreactive with said reagent, said first bibulous member being in liquid receiving relationship with a second bibulous member, under conditions wherein said second medium traverses said first bibulous member and at least a portion of said second bibulous member by capillary action, with the provision that said first bibulous member is in liquid receiving relationship with said absorbent member and said second bibulous member at substantially different times; and

determining a border on said second bibulous member, said border being defined by a signal produced by contacting said second bibulous member with a detection agent, the position of said border being related to the amount of analyte in said first medium or the amount of reagent or the amount of said component in said second liquid medium.

34. The method of claim 33 wherein said analyte is a drug, said reagent is antibody for said drug, and said component is enzyme labeled drug analog.

35. A method for determining the amount of an analyte, which comprises:

causing a reagent in a first liquid medium to become bound to a first bibulous member zone in relation to the presence or amount of analyte by contacting said first bibulous member zone with said medium under conditions wherein said first medium flows through or traverses said first bibulous member zone by capillary action;

causing a component in a second liquid medium to be absorbed by a second bibulous member zone in relation to the presence of said analyte in said first medium by contacting a portion of said first bibulous member with said second medium, under conditions wherein said second member transverse said first bibulous member zone and at least a portion of said second bibulous member zone by capillary action, sad second bibulous member zone having a binding partner for said component non-diffusively bound therein; and

determining the distance said component traverses said second bibulous member zone to determine the amount of said analyte, or determining the presence of said component on at least a portion of said second bibulous member to determine the presence of said analyte.

36. The method of claim 35 wherein said second bibulous member zone is an immunochromatographic member.

37. The method of claim 35 wherein a binding partner capable of binding said reagent is non-diffusively bound to at least a portion of said first bibulous member zone.

38. The method of claim 35 wherein said reagent and said analyte bind together to form a complex and said complex becomes bound to said first bibulous member zone.

39. The method of claim 35 where said analyte is an sbp member.