Simultaneous calibration heterogeneous immunoassay

• US 5,342,759 A
• Filed: 04/03/1989
• Issued: 08/30/1994
• Est. Priority Date: 12/26/1979
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Syva Co. (Siemens AG)
• Inventors: Ullman, Edwin F., Litman, David J.
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US07/333,745
• Time in Patent Office: 1,975 Days
• Field of Search: 422/56, 435/4, 435/7.92, 435/805, 435/967, 435/970, 435/973, 435/975, 436/37, 436/518
• US Class Current: 435/7.92
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/581 with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• 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 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 436/82 Hepatitis associated antigens and antibodies
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives

Abstract

An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) comprising ligand and receptor. By providing a first measurement surface capable of specifically binding a labelled reagent in an amount depending upon the presence of analyte in the sample and a second calibration surface capable of binding a second labeled reagent in a manner unaffected by the presence of analyte in the sample, calibration of individual tests can be accomplished simulataneously with the performance of the test itself. A signal producing system includes an enzyme bonded to a mip which defines the first labeled reagent for binding to the measurement surface and the same enzyme conjugated to a ligand capable of binding to the calibration surface. Preferably, both labeled reagents have the same composition and the calibration surface includes anti-(first enzyme).

Claims

1. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing at least one catalyst including a catalyst bound to a mip ("catalyst-bound-mip") and a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in proportion to the amount of catalyst-bound-mip bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-mip results in binding of said catalyst-bound-mip to said first surface in proportion to the amount of analyte in said sample,

the improvement which comprises

having adjacent to said first surface on a common underlying structure a calibration second surface to which said catalyst becomes bound, by the intermediacy of specific binding pair complex formation involving a determinant site other than that involved in the binding of the catalyst-bound-mip to said first surface, in an amount to produce a signal level at said second surface and said analyte in said sample is determined by comparing the intensity of the signal at said second surface to the intensity of the signal at said first surface.

2. A method according to claim 1, wherein said catalyst is an enzyme.

3. A method according to claim 1, wherein catalyst becomes bound to said second surface during said contacting by means of mip complex formation.

4. A method according to claim 3 wherein said mip complex formation involves anti-enzyme on said calibration surface.

5. A method according to claim 3 wherein said mip of said catalyst-bound-mip is a first receptor and said mip complex formation involves a second receptor for said first receptor on said calibration surface.

6. A method according to claim 5 wherein said second receptor is anti-(antibody).

7. A method according to claim 6 wherein said anti-(antibody) is an anti-idiotype antibody.

8. A method according to claim 1 wherein said catalyst-bound-mip is enzyme-bound-ligand.

9. A method according to claim 1 wherein said catalyst-bound-mip is enzyme-bound-antiligand.

10. An internally calibrated diagnostic apparatus for determining the presence or amount of an analyte in a sample, said apparatus comprising a support, a measurement first surface of a porous material mounted on said support;

a calibration second surface of a porous material mounted on said support in close proximity to said first surface;

a member of a specific binding pair (mip) non-diffusively bound to said first surface; and

at least one of an enzyme, anti-enzyme or receptor for a receptor for said mip non-diffusively bound to said second surface in an amount such that the difference in the detectable signal between the calibration second surface and the measurement first surface is qualitatively or quantitatively related to the amount of analyte in the sample.

11. A device according to claim 10, wherein both said first and second surfaces have the same enzyme non-diffusively bound to said surfaces.

12. A kit for use in a diagnostic assay comprising a device according to claim 10 and an enzyme-bound-mip, where the mip bound to said first surface and the mip of said enzyme-bound-mip are the same, homologous or both bind to different binding sites of the same ligand.

13. A method for conducting an assay for the presence or amount of an analyte selected from the group consisting of ligands and receptors in a liquid medium, which comprises:

subjecting an underlying structure comprising measurement and calibration surfaces to the same assay environment whereby a signal resulting from an enzyme on each of said surfaces is generated, said enzyme either being initially bound to said calibration surface or becoming bound to said calibration surface by the intermediacy of specific binding pair complex formation involving a determinant site other than that involved in the binding of the enzyme to said measurement surface, and

comparing the intensities of signals at said measurement and calibration surfaces to determine the presence or amount of said analyte in said liquid medium.

14. A method for conducting an assay for the presence or amount of an analyte, which comprises:

subjecting to the same assay environment (1) a measurement surface to which a catalyst-specific binding pair member ("mip") conjugate becomes bound in relation to the amount of an analyte in a liquid medium and (2) a calibration surface to which an amount of catalyst becomes bound, by the intermediacy of specific binding pair complex formation involving a determinant site other than that involved in the binding of the catalyst-mip conjugate to said measurement surface, that is substantially independent of the amount of analyte in said liquid medium whereby a signal is generated on each of said surfaces, said measurement surface and said calibration surface being on a common support, and

comparing the intensity of the signal at said measurement surface with the intensity of the signal at said calibration surface to determine the presence or amount of said analyte.

15. A method according to claim 14 wherein said calibration surface has a receptor for an enzyme catalyst bound thereto.

16. A method according to claim 15 wherein said receptor for an enzyme is an anti-enzyme.

17. A method according to claim 14 wherein said mip is an antibody and said calibration surface has anti-(antibody) bound thereto.

18. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing at least one catalyst including an enzyme catalyst bound to a mip ("catalyst-bound-mip") and a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in relation to the amount of catalyst-bound-mip bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-mip results in binding of said catalyst-bound-mip to said first surface in relation to the amount of analyte in said sample,

the improvement which comprises:

having adjacent to said first surface a calibration second surface to which a receptor for enzyme catalyst is bound in an amount to cause binding of such catalyst-bound-mip and produce a signal at said second surface and the presence or amount of said analyte in said sample is determined by comparing the intensity of said signal at said first surface to the intensity of said signal at said second surface.

19. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing (1) at least one catalyst including a catalyst bound to a mip ("catalyst-bound-mip") which mip is a first receptor and (2) a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in relation to the amount of catalyst-bound-mip bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-mip results in binding of said catalyst-bound-mip to said first surface in relation to the amount of analyte in said sample,

the improvement which comprises:

having adjacent to said first surface on a common underlying support a calibration second surface to which a second receptor for said first receptor is bound in an amount to cause binding of said catalyst-bound-mip and thereby produce a signal at said second surface and the presence or amount of said analyte in said sample is determined by comparing the intensity of said signal at said first surface to the intensity of said signal at said second surface.

20. A method according to claim 19, wherein said catalyst is an enzyme.

21. A method according to claim 19, wherein said first receptor is an antibody and said second receptor for said first receptor is an anti-(antibody).

22. A method according to claim 21, wherein said anti-(antibody) is an anti-idiotype antibody.

23. A method according to claim 21 wherein said anti-(antibody) binds to the Fc portion of the antibody.

24. A device for conducting an assay for the presence or amount of an analyte in a sample suspected of containing said analyte, which comprises:

a support comprising at least one measurement first surface of a porous material,

at least one calibration second surface of a porous material on said support in close proximity to said first surface,

a member of a specific binding pair non-diffusively bound to said first surface, and

a receptor non-diffusively bound to said second surface, said receptor being selected from the group consisting of a receptor for an enzyme, a receptor for a second receptor that is part of a conjugate of said second receptor and an enzyme, and a receptor for a calibration hapten,

said receptor being bound to said second surface in an amount such that the difference in a detectable signal between the second surface and the first surface is qualitatively or quantitatively related to the amount of analyte in the sample.

25. A device according to claim 24 wherein said receptor is an antibody.

26. A device according to claim 24 wherein said receptor is an anti-(antibody).

27. A device according to claim 24 wherein said receptor is an anti-idiotype antibody.

28. A device according to claim 24 comprising a plurality of one or both of said measurement and calibration surfaces.

29. A kit comprising the device of claim 24.

30. The kit of claim 29 comprising as a reagent separate from said device a conjugate of a second receptor and an enzyme.

31. The kit of claim 30 wherein said catalyst is an enzyme.

32. The kit of claim 30 when said mip is an antibody.

33. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing at least one enzyme catalyst including an enzyme catalyst bound to a mip ("catalyst-bound-mip") and a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in proportion to the amount of catalyst-bound-mip bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-mip results in binding of said catalyst-bound-mip to said first surface in proportion to the amount of analyte in said sample,

the improvement which comprises"

having adjacent to said first surface a calibration second surface to which said enzyme catalyst becomes bound, during said contacting by means of mip complex formation involving anti-enzyme catalyst on said calibration surface, in an amount to produce a signal level at said second surface and said analyte in said sample is determined by comparing the intensity of the signal at said second surface to the intensity of the signal at said first surface.

34. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing at least one catalyst including a catalyst bound to an antibody ("catalyst-bound-antibody") and a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in proportion to the amount of catalyst-bound-antibody bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-antibody results in binding of said catalyst-bound-antibody to said first surface in proportion to the amount of analyte in said sample,

the improvement which comprises"

having adjacent to said first surface a calibration second surface to which said catalyst becomes bound, during said contacting by means of mip complex formation involving an anti-(antibody) on said calibration surface, in an amount to produce a signal level at said second surface, and said analyte in said sample is determined by comparing the intensity of the signal at said second surface to the intensity of the signal at said first surface.

35. A method according to claim 34 wherein said anti-(antibody) is an anti-idiotype antibody.

36. A method for conducting an assay for the presence or amount of an analyte, which comprises:

subjecting to the same assay environment (1) a measurement surface to which a catalyst-specific binding pair member ("mip") conjugate becomes bound in relation to the amount of an analyte in a liquid medium and (2) a calibration surface having a receptor for an enzyme catalyst to which an amount of enzyme becomes bound that is substantially independent of the amount of analyte in said liquid medium whereby a signal is generated on each of said surfaces and

comparing the intensity of the signal at said measurement surface with the intensity of the signal at said calibration surface to determine the presence or amount of said analyte.

37. A method according to claim 36 wherein said receptor for an enzyme is an anti-enzyme.

38. A method for conducting an assay for the presence or amount of an analyte, which comprises:

subjecting to the same assay environment (1) a measurement surface to which a catalyst-antibody conjugate becomes bound in relation to the amount of an analyte in a liquid medium and (2) a calibration surface having an anti-(antibody) to which an amount of catalyst becomes bound that is substantially independent of the amount of analyte in said liquid medium whereby a signal is generated on each of said surfaces and

comparing the intensity of the signal at said measurement surface with the intensity of the signal at said calibration surface to determine the presence or amount of said analyte.

39. In a method for determining the presence or amount in a sample of an analyte, which analyte is a member of a specific binding pair ("mip") consisting of ligand and receptor ("antiligand"),

said method employing (1) at least one catalyst including a catalyst bound to a mip ("catalyst-bound-mip"), which mip is an antibody, and (2) a solute which is catalytically transformed by a catalyst bound to a mip-containing measurement first surface to produce a detectable signal at said first surface in relation to the amount of catalyst-bound-mip bound to said first surface, wherein contacting of said first surface with said sample and said catalyst-bound-mip results in binding of said catalyst-bound-mip to said first surface in relation to the amount of analyte in said sample,

the improvement which comprises:

having adjacent to said first surface a calibration second surface to which an anti-(antibody) is bound in an amount to cause binding of said catalyst-bound-mip and thereby produce a signal at said second surface and the presence or amount of said analyte in said sample is determined by comparing the intensity of said signal at said first surface to the intensity of said signal at said second surface.

40. A method according to claim 39 wherein said anti-(antibody) is anti-idiotype antibody.

41. A device for conducting an assay for the presence or amount of an analyte in a sample suspected of containing said analyte, which comprises:

at least one measurement first surface of a porous material,

at least one calibration second surface of a porous material in close proximity to said first surface,

a member of a specific binding pair non-diffusively bound to said first surface, and

an anti-(antibody) non-diffusively bound to said second surface, said anti-(antibody) binding to an antibody that is part of a conjugate of said antibody and an enzyme,

said anti-(antibody) being bound to said second surface in an amount such that the difference in a detectable signal between the second surface and the first surface is qualitatively or quantitatively related to the amount of analyte in the sample.