Assay method using internal calibration to measure the amount of analyte in a sample

• US 5,387,503 A
• Filed: 11/12/1992
• Issued: 02/07/1995
• Est. Priority Date: 06/06/1990
• Status: Expired due to Fees

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Selmer, Johan, Poulsen, Fritz
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US07/938,039
• Time in Patent Office: 817 Days
• Field of Search: 435/5, 435/7.2, 435/7.21, 435/7.23, 435/7.32, 435/7.4, 435/7.8, 435/7.94, 435/7.95, 435/967, 435/975, 436/501, 436/518, 436/807
• US Class Current: 435/5
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/96 involving blood or serum control standard
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

This invention relates to a method of determining the amount of test analyte in a sample using internal calibration comprising mixing a sample with a predetermined amount of a calibrator analyte foreign to the sample and with a comparable behavior in an assay to that of the test analyte, contacting the mixture with a solid support containing, each in a separate area, a regent for binding the test and calibrator analytes, respectively, contacting the solid support with a mixture of labelled reagents for binding the test and calibrator analytes, respectively, and determining the amount of test analyte in the sample by comparing the levels of labelled reagent bound to the test and calibrator analytes.

Claims

1. An assay for determining the amount of a test analyte contained in a sample, comprising the following sequential steps:

(a) providing a mixture of a known volume of said sample and a known amount of a calibrator analyte, wherein said calibrator analyte is foreign to said sample and the behavior of said calibrator analyte is comparable to that of said test analyte under the same assay conditions;

(b) contacting said mixture with a solid support, wherein said solid support comprises a first reagent capable of selectively binding said test analyte bound to a first discrete area of said solid support and a second reagent capable of selectively binding said calibrator analyte bound to a second discrete area of said solid support;

(c) contacting said solid support with a first labelled substance capable of selectively binding said test analyte and a second labelled substance capable of selectively binding said calibrator analyte, wherein said first and second labelled substances use an identical label; and

(d) determining the amount of said test analyte in said sample by comparing the level of said first labelled substance bound to said test analyte and of said second labelled substance bound to said calibrator analyte.

2. The assay according to claim 1, wherein said calibrator analyte is an animal or a plant protein which does not cross-react with said test analyte.

3. The assay according to claim 2, wherein said calibrator analyte is selected from the group consisting of aprotinin, a synthetic polypeptide, a hapten bound to a macromolecular compound, a polysaccharide, and an antibody.

4. The assay according to claim 2, wherein said test analyte is selected from the group consisting of a hormone, a blood component, a coagulation factor, a protease, a growth factor, a microbial component, a pancreatic amylase, a heart structural protein, a C-reactive protein, a α

-foetoprotein, a creatine kinase type MB, a creatine kinase type MM, a bacterial or a viral surface protein, and an antibody.

5. The assay according to claim 4, wherein said test analyte is a hormone selected from the group consisting of insulin, a growth hormone, a chorionic gonadotropin, a luteinizing hormone, a follicle stimulating hormone, and a steroid hormone.

6. The assay according to claim 4, wherein said test analyte is a blood component selected from the group consisting of a blood coagulation factor, a protease, and a precursor of a protease.

7. The assay according to claim 4, wherein said test analyte is a coagulation factor selected from the group consisting of Factor VII, Factor VIII, Factor IX, Factor X, and Factor XIII.

8. The assay according to claim 4, wherein the test analyte is a protease selected from the group consisting thrombin, prothrombin, plasmin, plasminogen, and tissue plasminogen activator.

9. The assay according to claim 4, wherein said test analyte is a growth factor selected from the group consisting of insulin-like growth factor, platelet-derived growth factor, epidermal growth factor, colony stimulating factor, transforming growth factor, and fibroblast growth factor.

10. The assay according to claim 4, wherein said test analyte is a microbial component selected from the group consisting of cholera toxin, diphtheria toxin, Salmonella toxin, Shiga toxin, Clostridium botulinum toxin, aflatoxin, endotoxin, and Chlamydia-specific antigen.

11. The assay according to claim 1, wherein said test analyte is selected from the group consisting of a creatine kinase type MB, a creatine kinase type MM, a pancreatic amylase, a heart structural protein, a C-reactive protein, a α

-foetoprotein, and a Chlamydia-specific antigen.

12. The assay according to claim 1, wherein said sample is selected from the group consisting of a body fluid, a tissue extract, a culture medium in which a cell has been grown, and a foodstuff.

13. The assay according to claim 12, wherein the body fluid is one selected from the group consisting of blood, serum, plasma, amniotic fluid, sputum, urine, cerebrospinal fluid, lymph, tear fluid, feces, and gastric fluid.

14. The assay according to claim 1, wherein said first reagent is selected from the group consisting of an antibody reactive with said test analyte, a cell surface receptor, a ligand-binding part of a cell surface receptor, a lectin, an antigen, and an anti-idiotype antibody.

15. The assay according to claim 14, wherein said first reagent is a monoclonal antibody or a polyclonal antibody.

16. The assay according to claim 14, wherein said first reagent is a glycoprotein or a glycolipid receptor.

17. The assay according to claim 14, wherein said first reagent is selected from the group consisting of lentil lectin, wheat germ, lectin, peanut lectin, soybean lectin and concanavalin A.

18. The assay according to claim 14, wherein said first reagent is a natural or synthetic antigen containing at least one epitope which is reactive with a test analyte antibody.

19. The assay according to claim 14, wherein said first reagent is an anti-idiotype antibody which is reactive with a test analyte antibody.

20. The assay according to claim 14, wherein said first reagent is a monoclonal or a polyclonal antibody.

21. The assay according to claim 1, wherein said second reagent is selected from the group consisting of an antibody reactive with the calibrator analyte, a cell surface receptor, a ligand-binding part a cell surface receptor, an antigen, a lectin, and an anti-idiotype antibody.

22. The assay according to claim 21, wherein said second reagent is a monoclonal or polyclonal antibody.

23. The assay according to claim 21, wherein said second reagent is a glycoprotein or a glycolipid receptor.

24. The assay according to claim 21, wherein said second reagent is a lectin selected from the group consisting of lentil lectin, wheat germ lectin, peanut lectin, soybean lectin, and concanavalin A.

25. The assay according to claim 21, wherein said second reagent is an anti-idiotype antibody which is reactive with a calibrator analyte antibody.

26. The assay according to claim 25, wherein said second reagent is a monoclonal or a polyclonal antibody.

27. The assay according to claim 1, wherein the label of said first and second labelled substances is selected from the group consisting of an enzyme, a colored substance, a fluorescent substance, and a radioactive isotope.

28. The assay according to claim 1, wherein said calibrator analyte is aprotinin and said second reagent is an anti-aprotinin antibody.

29. The assay according to claim 28, wherein said first labelled substance is an enzyme-labelled antibody, and said second labelled substance is an enzyme labelled anti-aprotinin antibody.

30. The assay according to claim 28, wherein said test analyte is selected from the group consisting of a creatine kinase type MB, a creatine kinase type MM, a pancreatic amylase, a heart structural protein, a C-reactive protein, a α

-foetoprotein, and a Chlamydia-specific antigen.

31. The assay according to claim 1, wherein said solid support further comprises an antibody nonreactive with either the test or calibrator analyte bound to a third discrete area of said solid support.

32. The assay according to claim 1, wherein said solid support further comprises one or more additional reagents capable of selectively binding to one or more additional test analytes bound to discrete areas of said solid support, and step (c) further comprises contacting said solid support with one or more additional labelled substances each capable of selectively binding said one or more additional test analytes, wherein said one or more additional labelled substances use the identical label.

33. A test kit for determining the amount of a test analyte in a sample, comprising

(a) a first container comprising a known amount of a calibrator analyte, wherein said calibrator analyte is foreign to said sample and the behavior of said calibrator analyte is comparable to that of said test analyte under the same assay conditions;

(b) a solid support comprising a first reagent capable of selectively binding said test analyte bound to a first discrete area of said solid support and a second reagent capable of selectively binding said calibrator analyte bound to a second discrete area of said solid support;

(c) a second container comprising a first labelled substance capable of selectively binding the test analyte; and

(d) a third container comprising a second labelled substance capable of selectively binding the calibrator analyte;

wherein said first and second labelled substances use an identical label.

34. The test kit according to claim 33, wherein the second and third containers are the same container.

35. The test kit according to claim 33, wherein said calibrator analyte is an animal or a plant protein which does not cross-react with the test analyte.

36. The test kit according to claim 35, wherein said calibrator analyte is selected from the group consisting of an aprotinin, a synthetic polypeptide, a hapten bound to a macromolecular compound, a polysaccharide, and an antibody.

37. The test kit according to claim 33, wherein said first reagent is selected from the group consisting of an antibody reactive with said test analyte, a cell surface receptor, a ligand-binding part of a cell surface receptor, a lectin, an antigen, and an anti-idiotype antibody.

38. The test kit according to claim 37, wherein said first reagent is a monoclonal or a polyclonal antibody or a fragment thereof.

39. The test kit according to claim 37, wherein said first reagent is a glycoprotein or a glycolipid receptor.

40. The test kit according to claim 37, wherein said first reagent is selected from the group consisting of lentil lectin, wheat germ lectin, peanut lectin, soybean lectin, and concanavalin A.

41. The test kit according to claim 37, wherein said first reagent is a natural or synthetic antigen containing at least one epitope which is reactive with a test analyte antibody.

42. The test kit according to claim 37, wherein said first reagent is an anti-idiotype antibody which is reactive with a test analyte antibody.

43. The test kit according to claim 33, wherein said second reagent is selected from the group consisting of an antibody reactive with said calibrator analyte, a cell surface receptor, a ligand-binding part of a cell surface receptor, an antigen, a lectin, and an anti-antibody.

44. The test kit according to claim 43, wherein said second reagent is a monoclonal or polyclonal antibody.

45. The test kit according to claim 43, wherein said second reagent is a glycoprotein or a glycolipid receptor.

46. The test kit according to claim 43, wherein said second reagent is selected from the group consisting of lentil lectin, wheat germ lectin, peanut lectin, soybean lectin, concanavalin A.

47. The test kit according to claim 43, wherein said second reagent is an anti-idiotype antibody which is reactive with a calibrator analyte antibody.

48. The test kit according to claim 47, wherein said second reagent is a monoclonal or polyclonal antibody or a fragment thereof.

49. The test kit according to claim 33, wherein said solid support is a polymer or a ceramic material.

50. The test kit according to claim 49, wherein said polymer is selected from the group consisting of plastic, a polysaccharide, an ion exchange resin, a cellulose polymer, and a silicon polymer.

51. The test kit according to claim 49, wherein said ceramic material is glass.

52. The test kit according to claim 49, wherein said solid support is selected from the group consisting of a plate, a paper strip, a dipstick, a membrane, and a solid particle.

53. The test kit according to claim 33, wherein said solid support further comprises an antibody nonreactive with either the test or calibrator analyte bound to a third discrete area of said solid support.

54. The test kit according to claim 33, wherein the label of said first and second labelled substances is selected from the group consisting of an enzyme, a colored substance, a fluorescent substance, and a radioactive isotope.

55. The test kit according to claim 33, further comprising one or more additional containers comprising one or more additional reagents capable of selectively binding to one or more additional test analytes bound to discrete areas of said solid support, and one or more additional containers comprising one or more additional labelled substances each capable of selectively binding said one or more additional test analytes, wherein said one or more additional labelled substances use the identical label.