Detection methods
View Patent ↗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.
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.
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.
6. The method of claim 1 for detecting an analyte A in a sample, wherein the method is a heterogeneous or a homogeneous sandwich test.
7. 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.
8. The method of claim 1 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.
9. The method of claim 1 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.
10. The method of claim 9 , wherein the suspendable solid phase comprises microparticles.
11. The method of claim 10 , wherein the microparticles function as a label.
12. 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.
13. The method of claim 12 , wherein the suspendable solid phase comprises microparticles.
14. The method of claim 13 , wherein the microparticles constitute the label L1.
15. 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.
16. The method of claim 15 , wherein the interaction comprises an energy transfer.
17. The method of claim 15 , wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.
18. 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.
19. The method of claim 18 for detecting an analyte A in a sample, wherein the method comprises at least one of detecting, avoiding, and decreasing a hook effect.
20. The method of claim 18 for detecting an analyte A in a sample, wherein the method comprises a heterogeneous or homogeneous sandwich test.
21. The method of claim 18 for detecting an analyte A in a sample, wherein the method comprises quantitatively or qualitatively detecting the analyte A in the sample.
22. 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.
23. 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.
24. The method of claim 5 for detecting an analyte A in a sample, wherein the method is a heterogeneous or a homogeneous sandwich test.
25. 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.
26. The method of claim 5 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.
27. The method of claim 5 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.
28. The method of claim 27 , wherein the suspendable solid phase comprises microparticles.
29. The method of claim 28 , wherein the microparticles function as a label.
30. 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.
31. The method of claim 30 , wherein the suspendable solid phase comprises microparticles.
32. The method of claim 31 , wherein the microparticles constitute the label L1.
33. 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.
34. The method of claim 33 , wherein the interaction comprises an energy transfer.
35. The method of claim 34 , wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.
36. The method of claim 18 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.
37. The method of claim 18 for detecting an analyte A in a sample, wherein L1 and L2 are the same label.
38. The method of claim 18 for detecting an analyte A in a sample, wherein the solid phase is a suspendable solid phase.
39. The method of claim 18 , wherein the suspendable solid phase comprises microparticles.
40. The method of claim 39 , wherein the microparticles function as a label.
41. The method of claim 18 for detecting an analyte A in a sample, wherein the binding partner R2 is associated with a suspendable solid phase.
42. The method of claim 41 , wherein the suspendable solid phase comprises microparticles.
43. The method of claim 42 , wherein the microparticles constitute the label L1.
44. The method of claim 18 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.
45. The method of claim 44 , wherein the interaction comprises an energy transfer.
46. The method of claim 45 , wherein the signal-forming system comprises photosensitizers which are associated with microparticles and chemiluminescent substances which are associated with microparticles.