Photoactivatable chemiluminescent matrices

• US 5,709,994 A
• Filed: 06/06/1995
• Issued: 01/20/1998
• Est. Priority Date: 07/31/1992
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Kirakossian, Hrair, Ullman, Edwin F., Pease, John S., Wagner, Daniel B.
• Primary Examiner(s): Myers, Carla J.
• Application Number:
  US08/470,862
• Time in Patent Office: 959 Days
• Field of Search: 435/6, 435/7.1, 435/7.92, 435/4, 435/810, 436/501, 436/3, 436/172, 436/805, 436/800, 436/829, 935/77, 935/78, 252/700, 252/582
• US Class Current: 435/4
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10S 436/905 Photochemical activation of reactions
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing

Abstract

Methods for labeling a material are disclosed. The methods comprise combining with the material (a) a photosensitizer capable upon irradiation of generating singlet oxygen and (b) a chemiluminescent compound capable of being activated by singlet oxygen wherein the photosensitizer and the chemiluminescent compound are incorporated in a particulate matrix or a non-particulate solid matrix. The particulate matrix can be solid or fluid. The methods allow for generating delayed luminescence, which can be realized upon irradiation of the matrix. The methods have application to the determination of an analyte in a medium suspected of containing the analyte. One method comprises subjecting a medium suspected of containing an analyte to conditions under which a complex of specific binding pair (sbp) members is formed in relation to the presence of the analyte and determining whether the sbp member complex has formed by employing as a label a single composition having both chemiluminescent and photosensitizer properties. Upon activation of the photosensitizer property singlet oxygen is generated and activates the chemiluminescent property. Compositions and kits are also disclosed.

Claims

1. A method for determining the presence or absence of an analyte, said method comprising:

irradiating a composition suspected of containing the analyte, said composition comprising a non-particulate solid matrix or a particulate matrix, said matrix having incorporated therein (1) a photosensitizer that upon irradiation generates singlet oxygen and (2) a chemiluminescent compound activatable by the singlet oxygen; and

determining the presence of the analyte.

2. The method of claim 1 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

3. A method for labeling a material, said method comprising:

combining with said material (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen, wherein said photosensitizer and said chemiluminescent compound are incorporated in a non-particulate solid matrix or a particulate matrix.

4. The method of claim 3 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

5. A method for detecting a material, said method comprising:

(a) combining with said material (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen, wherein said photosensitizer and said chemiluminescent compound are incorporated in a solid or particulate matrix;

(b) irradiating said material with light of wavelength 200-1000 nm;

(c) detecting light emitted by said chemiluminescent compound; and

(d) correlating the amount of detected light to the presence of the material.

6. The method of claim 5 wherein said material is selected from the group consisting of (a) an analyte, (b) a fluid, (c) an abraded material, (d) a cell, and (e) a biological tissue.

7. The method of claim 5 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

8. A method for generating delayed luminescence, said method comprising the step of irradiating a composition comprising a solid or particulate matrix having incorporated therein (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen.

9. The method of claim 8 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

10. The method of claim 8 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, phthalocyanines, and chlorophyll.

11. The method of claim 8 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

12. The method of claim 8 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

13. A method for determining the presence of an analyte, comprising the steps of:

subjecting a medium suspected of containing an analyte to conditions under which a complex of specific binding pair (sbp) members is formed in relation to the presence of said analyte;

determining whether said sbp member complex has formed by employing as a label a single composition having both chemiluminescent and photosensitizer properties such that upon activation of said photosensitizer property singlet oxygen is generated and activates said chemiluminescent property; and

determining the presence of said analyte.

14. The method of claim 13, wherein said single composition is a solid matrix or a particle having incorporated therein a photosensitizer that upon irradiation generates singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen.

15. The method of claim 13 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

16. The method of claim 13 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, chlorophyll and phthalocyanines.

17. The method of claim 13 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

18. The method of claim 13 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

19. A method for determining an analyte, said method comprising:

providing in combination (1) a medium suspected of containing said analyte, (2) a label reagent comprising a first specific binding pair (sbp) member associated with a single composition having both photosensitizer and chemiluminescent properties such that upon activation of said photosensitizer property singlet oxygen is generated and activates said chemiluminescent property, wherein said first sbp member binds to said analyte or to a second sbp member to form a complex related to the presence of said analyte, activating said photosensitizer property and detecting the amount of luminescence generated by said chemiluminescent property, the amount thereof being related to the amount of analyte in said medium.

20. The method of claim 19, wherein said single composition is a solid matrix or a particle having incorporated therein a photosensitizer that upon irradiation generates singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen.

21. The method of claim 20 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

22. The method of claim 20 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, chlorophyll and phthalocyanines.

23. The method of claim 20 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

24. The method of claim 20 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

25. A method for determining the presence of an analyte, said method comprising:

providing in combination (1) a medium suspected of containing said analyte, (2) a label reagent comprising a first specific binding pair (sbp) member associated with a particle having a photosensitizer that upon irradiation generates singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen such that, upon irradiating said photosensitizer, singlet oxygen is generated and activates said chemiluminescent compound, wherein said first sbp member binds to said analyte or to a second sbp member to form a complex related to the presence of said analyte;

irradiating said photosensitizer; and

detecting the amount of luminescence generated by the chemiluminescent compound, the amount thereof being related to the amount of analyte in said medium.

26. The method of claim 25 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

27. The method of claim 25 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, chlorophyll and phthalocyanines.

28. The method of claim 25 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

29. The method of claim 25 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

30. A composition comprising a solid matrix having incorporated therein a photosensitizer that upon activation generates singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen.

31. The composition of claim 30 wherein said photosensitizer is bound to said chemiluminescent compound.

32. The composition of claim 30 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

33. The composition of claim 30 comprising a plurality of distinct chemiluminescent compounds.

34. The composition of claim 33 wherein said distinct chemiluminescent compounds differ by differing rates of decay of emission following activation by singlet oxygen.

35. The composition of claim 30 wherein said photosensitizer and said chemiluminescent compound are covalently linked to said matrix.

36. The composition of claim 30 which comprises an activator that enhances the decay of activated chemiluminescent compound.

37. The composition of claim 30 comprising a member of a specific binding pair (sbp) bound thereto.

38. The composition of claim 37 wherein said sbp member is selected from the group consisting of ligands, receptors, polynucleotides and polynucleotide binding agents.

39. The composition of claim 30 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, chlorophyll and phthalocyanines.

40. The composition of claim 30 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

41. The composition of claim 30 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

42. The composition of claim 30 wherein said solid matrix is a particle having as average diameter of about 20 nanometers to 20 microns.

66. A kit comprising:

(a) the composition of claim 36 and

(b) a member of a specific binding pair.

43. A composition comprising a particle having incorporated therein a photosensitizer that generates singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen, said particle being bound to a molecule useful in the detection of an analyte.

44. The composition of claim 43 wherein said molecule is a member of a specific binding pair.

45. The composition of claim 43 wherein said photosensitizer is covalently bound to said chemiluminescent compound.

46. The composition of claim 44 wherein said sbp member is selected from the group consisting of ligands, receptors, polynucleotides and polynucleotide binding agents.

47. The composition of claim 43 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrins, chlorophyll, and phthalocyanines.

48. The composition of claim 43 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

49. The composition of claim 43 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

50. The composition of claim 43 wherein said photosensitizer and said chemiluminescent compound are dissolved in said particle.

51. The composition of claim 43 comprising a plurality of distinct chemiluminescent compounds.

52. The composition of claim 51 wherein said distinct chemiluminescent compounds differ by differing rates of decay after activation by singlet oxygen.

53. The composition of claim 43 which comprises an activator that enhances the decay of activated chemiluminescent compounds.

67. A kit comprising:

(a) the composition of claim 43 and

(b) a member of a specific binding pair.

54. A composition comprising fluid particles selected from the group consisting of oil droplets, liposomes and emulsions having incorporated therein a photosensitizer that upon activation of generating singlet oxygen and a chemiluminescent compound activatable by the singlet oxygen.

55. The composition of claim 54 wherein said photosensitizer is bound to said chemiluminescent compound.

56. The composition of claim 54 wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

57. The composition of claim 54 comprising a plurality of distinct chemiluminescent compounds.

58. The composition of claim 57 wherein said distinct chemiluminescent compounds differ by differing rates of decay of emission following activation by singlet oxygen.

59. The composition of claim 54 wherein said photosensitizer and said chemiluminescent compound are covalently linked to molecules comprising said fluid particles.

60. The composition of claim 54 which comprises an activator that enhances the decay of activated chemiluminescent compound.

61. The composition of claim 54 comprising a member of a specific binding pair (sbp) bound thereto.

62. The composition of claim 61 wherein said sbp member is selected from the group consisting of ligands, receptors, polynucleotides and polynucleotide binding agents.

63. The composition of claim 54 wherein said photosensitizer is a dye selected from the group consisting of methylene blue, rose bengal, porphyrin, chlorophyll and phthalocyanines.

64. The composition of claim 54 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin.

65. The composition of claim 54 wherein said chemiluminescent compound is selected from the group consisting of 9-alkylidene-N-methyl acridans, enol ethers and enamines.

68. A kit comprising:

(a) the composition of claim 55 and

(b) a member of a specific binding pair.

69. A method for determining a leak in a fluidic system, said method comprising:

introducing into a fluid in the fluidic system a composition comprising a non-particulate solid matrix or a particulate matrix, said matrix having incorporated therein (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen;

irradiating the fluid to activate the photosensitizer;

determining the presence of a leak by examining the system to determine if any luminescence is escaping from the system.

70. The method of claim 69, wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

71. A method for determining wear in a mechanical pad comprising:

incorporating into the mechanical part a composition comprising a non-particulate solid matrix or a particulate matrix, said matrix having incorporated therein (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen;

determining wear in the part by irradiating the part and detecting the presence of luminescence.

72. The method of claim 71, wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.

73. A method for detecting the emission of light comprising:

irradiating a composition comprising a non-particulate solid matrix or a particulate matrix, said matrix having incorporated therein (1) a photosensitizer that upon irradiation generates singlet oxygen, and (2) a chemiluminescent compound activatable by the singlet oxygen; and

detecting light emitted by said chemiluminescent compound.

74. The method of claim 73, wherein at least one of said photosensitizer and said chemiluminescent compound is covalently linked to said matrix.