Apparatus and method for homogeneous immunoassay

• US 4,537,861 A
• Filed: 02/03/1983
• Issued: 08/27/1985
• Est. Priority Date: 02/03/1983
• Status: Expired due to Term

• Current Assignee: Nicoli, David F., Virgil B. Elings
• Original Assignee: Nicoli, David F., Virgil B. Elings
• Inventors: Nicoli, David F., Elings, Virgil B.
• Primary Examiner(s): Marantz, Sidney
• Assistant Examiner(s): Wieder, Stephen C.
• Application Number:
  US06/463,658
• Time in Patent Office: 936 Days
• Field of Search: 250/461.1, 250/461.2, 356/317, 356/318, 356/417, 436/517, 436/518, 436/528, 436/819, 436/805, 436/807
• US Class Current: 436/518
• CPC Class Codes: G01N 2015/0222 from dynamic light scattering, e.g. photon correlation spectroscopy
• CPC Class Codes: G01N 2021/752 Devices comprising reaction zones
• CPC Class Codes: G01N 21/17 Systems in which incident light is modified in accordance with the properties of the material investigated where the material investigated is optically excited causing a change in wavelength of the incident light G01N21/63
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

Apparatus and method for an immunoassay of a binding reaction between a ligand and an antiligand which are typically an antigen and an antibody, including a spatial pattern formed by a spatial array of separate regions of antiligand material, and ligand material dispersed to interact with the spatial array of separate regions of antiligand material for producing a binding reaction between the ligand and the antiligand in the spatial patterns and with the bound complexes labeled with a particular physical characteristic. A source of input energy and with the input energy at a particular spectrum for interacting with particular physical characteristic of the labeled binding reaction. Scanning the spatial pattern with the input energy at the particular spectrum for producing output energy having amplitude levels formed by a substantially random background component and a non-random component representing the labeled bound complexes, and the non-random component representing the labeled bound complexes detected to produce an output signal in accordance with the labeled binding reaction.

Claims

1. Apparatus for providing an immunoassay of a binding reaction between an ligand and an antiligand, including

a spatial pattern formed by a predetermined known spatially periodic array of stripes of antiligand material and with the stripes having a predetermined width and spacing,

ligand material dispersed to interact with the spatially periodic array of stripes of antiligand material for producing a binding reaction between the ligand and the antiligand in the spatial pattern of stripes,

means for labeling the binding reaction with a particular physical characteristic,

a source of input energy and with the input energy at a particular spectrum for interacting with the particular physical characteristic of the labeled binding reaction,

means for scanning the spatial pattern with the input energy at the particular spectrum for producing output energy having amplitude levels formed by a substantially random background component and a periodic component representing the labeled binding reaction, and

means responsive to the output energy for detecting the periodic component representing the labeled binding reaction and for producing an output signal in accordance with the labeled binding reaction.

2. The apparatus for providing an immunoassay of claim 1 additionally including a cylindrical member and with the spatial pattern formed as a plurality of vertical stripes arranged circumferentially around the cylindrical member.

3. The apparatus for providing an immunoassay of claim 2 wherein the plurality of vertical stripes are on the inside surface of the cylindrical member and the ligand material is dispersed in a liquid contained within the cylindrical member.

4. The apparatus for providing an immunoassay of claim 2 wherein the means for scanning includes means for rotating the cylindrical member while the input energy is directed toward the spatial pattern.

5. The apparatus for providing an immunoassay of claim 1 additionally including a disc member and with the spatial pattern formed as a plurality of radial stripes arranged around a center point of the disc.

6. The apparatus for providing an immunoassay of claim 5 wherein the radial stripes are on a top surface of the disc and the ligand material is dispersed in a layer of liquid on the top surface of the disc.

7. The apparatus for providing an immunoassay of claim 5 wherein the means for scanning includes rotating the disc member while the input energy is directed toward the spatial pattern.

8. The apparatus for providing an immunoassay of claim 1 additionally including a substantially flat member and with the spatial pattern formed as a plurality of horizontal stripes arranged across the flat member.

9. The apparatus for providing an immunoassay of claim 8 wherein the horizontal stripes are on a top surface of the flat member and the ligand material is dispersed in a layer of liquid on the top surface of the flat member.

10. The apparatus for providing an immunoassay of claim 8 wherein the means for scanning includes scanning the input energy back and forth across the flat member in a direction perpendicular to the horizontal stripes.

11. The apparatus for providing an immunoassay of claim 1 wherein the labeled binding reaction between the ligand and antiligand is performed as a competitive assay including a known labeled ligand and an unknown unlabeled ligand both interacting with the antiligand to produce the binding reaction.

12. The apparatus for providing an immunoassay of claim 11 wherein the ligand to be labeled is attached to labeled carrier particles.

13. The apparatus for providing an immunoassay of claim 1 wherein the labeled binding reaction between the ligand and antiligand is performed as a sandwich assay including a known labeled antiligand and a known unlabeled antiligand and with an initial binding reaction between the ligand and the known unlabeled antiligand of a quantity sufficient to bind substantially all of the ligand and with a subsequent binding reaction between the known labeled antiligand and the product of the initial binding reaction.

14. The apparatus for providing an immunoassay of claim 13 wherein the antiligand to be labeled is attached to labeled carrier particles.

15. The apparatus for providing an immunoassay of claim 1 wherein the labeling of the binding reaction is a fluorescent label.

16. The apparatus for providing an immunoassay of claim 1 wherein the means for scanning includes moving the spatial pattern while the input energy is directed toward the moving spatial pattern.

17. The apparatus for providing an immunoassay of claim 1 wherein the means for scanning includes maintaining the spatial pattern stationary while scanning the input energy across the stationary spatial pattern.

18. The apparatus for providing an immunoassay of claim 1 wherein the means for detecting includes an electronic filter having a frequency range for passing the periodic component while discriminating against the random background component.

19. The apparatus for providing an immunoassay of claim 1 wherein the detecting includes a phase-lock detector for amplifying the periodic component while discriminating against the background component.

20. The apparatus for providing an immunoassay of claim 1 wherein the means for detecting includes an autocorrelator for extracting the periodic component from the random background component.

21. The apparatus for providing an immunoassay of claim 1 where the antiligand is attached to carrier particles and the periodic pattern is formed by a spatially periodic force field to which the carrier particles are responsive.

22. The apparatus for providing an immunoassay of claim 21 wherein the carrier particle is magnetic and the force field is a magnetic field.

23. The apparatus for providing an immunoassay of claim 21 wherein the carrier particle is electrically charged and the force field is an electric field.

24. The apparatus for providing an immunoassay of claim 21 wherein the carrier particle is electrically polarizable and the force field is an electric field.

25. Apparatus for providing an immunoassay of a binding reaction between a ligand and an antiligand, including

a spatial pattern formed by a predetermined known spatial array of separate areas of antiligand material,

ligand material dispersed to interact with the spatial array of separate areas of antiligand material for producing a binding reaction between the ligand and the antiligand in the spatial pattern,

means for labeling the binding reaction with a particular physical characteristic,

a source of input energy and with the input energy at a particular spectrum for interacting with the particular physical characteristic of the labeled binding reaction,

means for scanning the spatial pattern with the input energy at the particular spectrum for producing output energy having amplitude levels formed by a substantially random background component and a non-random component representing the labeled binding reaction, and

means responsive to the output energy for detecting the non-random component representing the labeled binding reaction and for producing an output signal in accordance with the labeled binding reaction.

26. The apparatus for providing an immunoassay of claim 25 additionally including a cylindrical member and with the spatial pattern formed as a plurality of vertical areas arranged circumferentially around the cylindrical member.

27. The apparatus for providing an immunoassay of claim 26 wherein the plurality of vertical areas are on the inside surface of the cylindrical member and the antigen material is dispersed in a liquid contained within the cylindrical member.

28. The apparatus for providing an immunoassay of claim 26 wherein the means for scanning includes means for rotating the cylindrical member while the input energy is directed toward the spatial pattern.

29. The apparatus for providing an immunoassay of claim 25 additionally including a disc member and with the spatial pattern formed as a plurality of radial areas arranged around a center point of the disc.

30. The apparatus for providing an immunoassay of claim 29 wherein the radial areas are on a top surface of the disc and the antigen material is dispersed in a layer of liquid on the top surface of the disc.

31. The apparatus for providing an immunoassay of claim 29 wherein the means for scanning includes rotating the disc member while the input energy is directed toward the spatial pattern.

32. The apparatus for providing an immunoassay of claim 25 additionally including a substantially flat member and with the spatial pattern formed as a plurality of horizontal areas arranged across the flat member.

33. The apparatus for providing an immunoassay of claim 32 wherein the horizontal areas are on a top surface of the flat member and the ligand material is dispersed in a layer of liquid on the top surface of the flat member.

34. The apparatus for providing an immunoassay of claim 32 wherein the means for scanning includes scanning the input energy back and forth across the flat member in a direction perpendicular to the horizontal stripes.

35. The apparatus for providing an immunoassay of claim 25 wherein the labeled binding reaction between the ligand and antiligand is performed as a competitive assay including a known labeled ligand and an unknown unlabeled ligand both interacting with the antiligand to produce the binding reaction.

36. The apparatus for providing an immunoassay of claim 35 wherein the ligand to be labeled is attached to labeled carrier particles.

37. The apparatus for providing an immunoassay of claim 25 wherein the labeled binding reaction between the ligand and antiligand is performed as a sandwich assay including a known labeled antiligand and a known unlabeled antiligand and with an initial binding reaction between the ligand and the known unlabeled antiligand of a quantity sufficient to bind all of the ligand and with a subsequent binding reaction between the known labeled antiligand and the product of the initial binding reaction.

38. The apparatus for providing an immunoassay of claim 37 wherein the antiligand to be labeled is attached to labeled carrier particles.

39. The apparatus for providing an immunoassay of claim 25 wherein the labeling of the binding reaction is a fluorescent label.

40. The apparatus for providing an immunoassay of claim 25 wherein the means for scanning includes moving the spatial pattern while the input energy is directed toward the moving spatial pattern.

41. The apparatus for providing an immunoassay of claim 25 wherein the means for scanning includes maintaining the spatial pattern stationary while scanning the input energy across the stationary spatial pattern.

42. The apparatus for providing an immunoassay of claim 25 wherein the means for detecting includes a filter having a frequency range for passing the non-random component while discriminating against the random background component.

43. The apparatus for providing an immunoassay of claim 25 wherein the means for detecting includes a phase lock detector for amplifying the non-random component while discriminating against the random background component.

44. The apparatus for providing an immunoassay of claim 25 wherein the means for detecting includes an autocorrelator for extracting the non-random component from the random background component.

45. The apparatus for providing an immunoassay of claim 25 where the antiligand is attached to carrier particles and the spatial pattern is formed by a spatial force field to which the carrier particles are responsive.

46. The apparatus for providing an immunoassay of claim 45 wherein the carrier particle is magnetic and the force field is a magnetic field.

47. The apparatus for providing an immunoassay of claim 45 wherein the carrier particle is electrically charged and the force field is an electric field.

48. The apparatus for providing an immunoassay of claim 45 wherein the carrier is electrically polarizable and the force field is an electric field.

49. A method for providing an immunoassay of a binding reaction between a ligand and an antiligand including the following steps,

providing a spatial pattern formed by a predetermined known spatial array of separate areas of antiligand material,

providing a ligand material dispersed to interact with the spatial array of separate areas of antiligand material for producing a binding reaction between the ligand and the antiligand in the spatial pattern,

labeling the binding reaction with a particular physical characteristic,

providing input energy at a particular spectrum for interacting with the paticular physical characteristic of the labeled binding reaction,

scanning the spatial pattern with the input energy at the particular spectrum for producing output energy having amplitude levels formed by a substantially random background component and a non-random component representing the labeled binding reaction, and

detecting the non-random component representing the labeled binding reaction and for producing an output signal in accordance with the labeled binding reaction.

50. The method of claim 49 wherein the spatial pattern is formed as a plurality of vertical areas arranged circumferentially around a cylindrical member.

51. The method of claim 50 wherein the plurality of vertical areas are on the inside surface of the cylindrical member and the antigen material is dispersed in a liquid contained within the cylindrical member.

52. The method of claim 50 wherein the scanning includes rotating the cylindrical member while the input energy is directed toward the spatial pattern.

53. The method of claim 49 wherein the spatial pattern is formed as a plurality of radial areas arranged around a center point of the disc.

54. The method of claim 53 wherein the radial areas are on a top surface of the disc and the antigen material is dispersed in a layer of liquid on the top surface of the disc.

55. The method of claim 53 wherein the scanning includes rotating the disc member while the input energy is directed toward the spatial pattern.

56. The method of claim 49 wherein the spatial pattern is formed as a pluraity of horizontal areas arranged across the flat member.

57. The method of claim 56 wherein the horizontal areas are on a top surface of the flat member and the ligand material is dispersed in a layer of liquid on the top surface of the flat member.

58. The method of claim 56 wherein the scanning includes scanning the input energy back and forth across the flat member in a direction perpendicular to the horizontal stripes.

59. The method of claim 49 wherein the labeled binding reaction between the ligand and antiligand is performed as a competitive assay including a known labeled ligand and an unknown unlabeled ligand both interacting with the antiligand to the binding reaction.

60. The method of claim 59 wherein the ligand to be labeled is attached to labeled carrier particles.

61. The method of claim 49 wherein the labeled binding reaction between the ligand and antiligand is performed as a sandwich assay including a known labeled antiligand and a known unlabeled antiligand and with an initial binding reaction between the ligand and the known unlabeled antiligand of a quantity sufficient to bind all of the ligand and with a subsequent binding reaction betwen the known labeled antiligand and the product of the initial binding reaction.

62. The method of claim 61 wherein the antiligand to be labeled is attached to labeled carrier particles.

63. The method of claim 49 wherein the labeling of the binding reaction is a fluorescent label.

64. The method of claim 49 wherein the scanning includes moving the spatial pattern while the input energy is directed toward the moving spatial pattern.

65. The method of claim 49 wherein the scanning includes maintaining the spatial pattern stationary while scanning the input energy across the stationary spatial pattern.

66. The method of claim 49 wherein the detecting includes filtering with a frequency range for passing the non-random component while discriminationg against the random background component.

67. The method of claim 49 wherein the detecting includes phase-lock detecting by amplifying the non-random component while discriminating against the random background component.