DETECTION METHODS

• US 20100311185A1
• Filed: 12/21/2001
• Published: 12/09/2010
• Est. Priority Date: 12/22/2000
• Status: Active Grant

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Inventors: Schelp, Carsten, Pauly, Hans-Erwin
• Granted Patent: US 7,867,781 B2
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: G01N 2470/04 Sandwich assay format
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms

Abstract

The invention relates to a method for quantitatively or qualitatively detecting an analyte in a sample, with the sample being incubated, for the purpose of avoiding, diminishing and/or detecting the high-dose hook effect, with an analyte-specific binding partner R1, which is associated with a solid phase, an analyte-specific binding partner R2, which is associated with a label L1, and an analyte-specific binding partner R3, which is associated with a label L2, and the L1-dependent measurement signal being determined either at a different time from the L2-dependent or L1 plus L2-dependent measurement signal or using a different measurement method.

Claims

1. A method for detecting an analyte A in a sample, comprising:

(i) incubating an incubation mixture comprising a sample with an analyte A-specific binding partner R1, which is associated with a solid phase, an analyte A-specific binding partner R2, which is associated with a label L1, and an analyte A-specific binding partner R3, which is associated with a label L2, wherein binding partners R2 and R3 are selected such that saturation of analyte A-binding sites of the binding partner R2 requires a) a higher analyte A concentration, b) a longer incubation, or c) a higher analyte A concentration and a longer incubation, than does saturation of analyte A-binding sites of the binding partner R3; and

(ii) determining an L1-dependent measurement signal at time T1 and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal at time T2, wherein time T1 is earlier than time T2, time T2 is after addition of label L2, and time T1 occurs before 30% of time from addition of label L2 to time T2 has elapsed;

or determining an L1-dependent measurement signal using a first measurement method and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal using a second measurement method, wherein the first and second measurement methods are different.

2. The method of claim 1 for detecting an analyte A in a sample, wherein the method comprises a quantitative measurement.

3. The method of claim 1 for detecting an analyte A in a sample, wherein the method comprises a qualitative measurement.

4. The method of claim 1 for detecting an analyte A in a sample, wherein the method comprises at least one of detecting, avoiding, and decreasing a hook effect.

7. The method of claim 1 for detecting an analyte A in a sample, wherein the method is a heterogeneous or a homogeneous sandwich test.

8. The method of claim 1 for detecting an analyte A in a sample, wherein R1 and R2;

R1 and R3;

R1, R2, and R3;

or R2 and R3 are the same binding partner.

9. The method of claim 1 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.

10. The method of claim 1 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.

11. The method of claim 10, wherein the suspendable solid phase comprises microparticles.

12. The method of claim 11, wherein the microparticles function as a label.

13. The method of claim 1 for detecting an analyte A in a sample, wherein the binding partner R2 is associated with a suspendable solid phase.

14. The method of claim 13, wherein the suspendable solid phase comprises microparticles.

15. The method of claim 14, wherein the microparticles constitute the label L1.

16. The method of claim 1 for detecting an analyte A in a sample, wherein, as a consequence of formation of a sandwich, components of a signal-forming system, which include at least one of L1 and L2, are brought to a distance from each other which permits an interaction between these components, and the extent of the interaction is measured.

17. The method of claim 16, wherein the interaction comprises an energy transfer.

18. The method of claim 16, wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.

5. A method for detecting an analyte A in a sample, comprising:

(i) incubating an incubation mixture comprising a sample with an analyte A-specific binding partner R1, which is associated with a solid phase, an analyte A-specific binding partner R2, which is associated with a label L1, and an analyte A-specific binding partner R3, which is associated with a first member of a specific binding pair, wherein binding partners R2 and R3 are selected such that saturation of analyte A-binding sites of the binding partner R2 requires a) a higher analyte A concentration, b) a longer incubation, or c) a higher analyte A concentration and a longer incubation, than does saturation of analyte A-binding sites of the binding partner R3;

(ii) incubating the incubation mixture of (i) with a label L2, which is associated with a second member of the specific binding pair; and

(iii) determining an L1-dependent measurement signal at time T1 and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal at time T2, wherein time T1 is earlier than time T2, time T2 is after addition of label L2, and time T1 occurs before 30% of time from addition of label L2 to time T2 has elapsed;

or determining an L1-dependent measurement signal using a first measurement method and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal using a second measurement method, wherein the first and second measurement methods are different.

32. The method of claim 5 for detecting an analyte A in a sample, wherein the method comprises quantitatively or qualitatively detecting the analyte A in the sample.

33. The method of claim 5 for detecting an analyte A in a sample, wherein the method comprises at least one of detecting, avoiding, and decreasing a hook effect.

34. The method of claim 5 for detecting an analyte A in a sample, wherein the method is a heterogeneous or a homogeneous sandwich test.

35. The method of claim 5 for detecting an analyte A in a sample, wherein R1 and R2;

R1 and R3;

R1, R2, and R3;

or R2 and R3 are the same binding partner.

36. The method of claim 5 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.

37. The method of claim 5 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.

38. The method of claim 37, wherein the suspendable solid phase comprises microparticles.

39. The method of claim 38, wherein the microparticles function as a label.

40. The method of claim 5 for detecting an analyte A in a sample, wherein the binding partner R2 is associated with a suspendable solid phase.

41. The method of claim 40, wherein the suspendable solid phase comprises microparticles.

42. The method of claim 41, wherein the microparticles constitute the label L1.

43. The method of claim 5 for detecting an analyte A in a sample, wherein, as a consequence of formation of a sandwich, components of a signal-forming system, which include at least one of L1 and L2, are brought to a distance from each other which permits an interaction between these components, and the extent of the interaction is measured.

44. The method of claim 43, wherein the interaction comprises an energy transfer.

45. The method of claim 44, wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.

6. (canceled)

19. A method for detecting an analyte A in a sample, comprising:

(i) incubating an incubation mixture comprising a sample with an analyte A-specific binding partner R1, which is associated with a solid phase, an analyte A-specific binding partner R2, which is associated with a label L1, and an analyte A-specific binding partner R3, which is associated with a first member of a specific binding pair, and a label L2, which is associated with a second member of the specific binding pair, wherein binding partners R2 and R3 are selected such that saturation of analyte A-binding sites of the binding partner R2 requires a) a higher analyte A concentration, b) a longer incubation, or c) a higher analyte A concentration and a longer incubation, than does saturation of analyte A-binding sites of the binding partner R3; and

(ii) determining an L1-dependent measurement signal at time T1 and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal at time T2, wherein time T1 is earlier than time T2, time T2 is after addition of label L2, and time T1 occurs before 30% of time from addition of label L2 to time T2 has elapsed;

or determining an L1-dependent measurement signal using a first measurement method and an L2-dependent measurement signal or an L1 plus L2-dependent measurement signal using a second measurement method, wherein the first and second measurement methods are different.

20. The method of claim 19 for detecting an analyte A in a sample, wherein the method comprises at least one of detecting, avoiding, and decreasing a hook effect.

21. The method of claim 19 for detecting an analyte A in a sample, wherein the method comprises a heterogeneous or homogeneous sandwich test.

22. The method of claim 19 for detecting an analyte A in a sample, wherein the method comprises quantitatively or qualitatively detecting the analyte A in the sample.

46. The method of claim 19 for detecting an analyte A in a sample, wherein R1 and R2;

R1 and R3;

R1, R2, and R3;

or R2 and R3 are the same binding partner.

47. The method of claim 19 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.

48. The method of claim 19 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.

49. The method of claim 48, wherein the suspendable solid phase comprises microparticles.

50. The method of claim 49, wherein the microparticles function as a label.

51. The method of claim 19 for detecting an analyte A in a sample, wherein the binding partner R2 is associated with a suspendable solid phase.

52. The method of claim 51, wherein the suspendable solid phase comprises microparticles.

53. The method of claim 52, wherein the microparticles constitute the label L1.

54. The method of claim 19 for detecting an analyte A in a sample, wherein, as a consequence of formation of a sandwich, components of a signal-forming system, which include at least one of L1 and L2, are brought to a distance from each other which permits an interaction between these components, and the extent of the interaction is measured.

55. The method of claim 54, wherein the interaction comprises an energy transfer.

56. The method of claim 55, wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.

23. A test kit for detecting an analyte A in a sample, comprising:

an analyte A-specific binding partner R1, which is associated with a solid phase;

an analyte A-specific binding partner R2, which is associated with a label L1; and

an analyte A-specific binding partner R3, which is associated with a label L2;

wherein saturation, in an incubation mixture of a sandwich test, of analyte A-binding sites of the binding partner R2 takes place at a higher analyte A concentration, at a later time in the incubation, or at a higher analyte A concentration and at a later time in the incubation, than does saturation of analyte A-binding sites of the binding partner R3.

24. The test kit of claim 23, wherein the test kit comprises a heterogeneous sandwich test kit or a homogeneous sandwich test kit.

25. The test kit of claim 23, wherein the test kit comprises a quantitative measurement test kit or a qualitative measurement test kit.

26. The test kit of claim 23, wherein the analyte A-specific binding partners R1, R2, and R3 are in separate receptacles.

27. A test kit for detecting an analyte A in a sample, comprising:

an analyte A-specific binding partner R1, which is associated with a solid phase;

an analyte A-specific binding partner R2, which is associated with a label L1;

an analyte A-specific binding partner R3, which is associated with a member X of a specific binding pair; and

a label L2, which is associated with the binding pair member Y, corresponding to X, of the specific binding pair;

wherein saturation, in an incubation mixture of a sandwich test, of analyte A-binding sites of the binding partner R2 takes place at a higher analyte A concentration, at a later time in the incubation, or at a higher analyte A concentration and at a later time in the incubation, than does saturation of analyte A-binding sites of the binding partner R3.

28. The test kit of claim 27, wherein the test kit comprises a heterogeneous sandwich test kit or a homogeneous sandwich test kit.

29. The test kit of claim 27, wherein the test kit comprises a quantitative measurement test kit or a qualitative measurement test kit.

30. The test kit of claim 27, wherein the analyte A-specific binding partners R1, R2, and R3 are in separate receptacles.

31. The test kit of claim 27, wherein the analyte A-specific binding partner R2, which is associated with the label L1, and the analyte A-specific binding partner R3, which is associated with the member X of the specific binding pair, are present together in one receptacle.