Assay particles and methods of use
View Patent ↗The invention provides assay particles useful, for example, for detecting analytes and binding molecule interactions. One type of assay particle includes a core portion encased by a shell portion, wherein the shell portion comprises an inorganic phosphor that binds selectively to a target molecule. Another type of an assay particle includes a core portion encased by a shell portion, and a coat portion covering the shell portion, wherein the coat portion comprises an inorganic phosphor that binds selectively to a target molecule. A further type of assay particle includes a core portion encased by a shell portion, and a coat portion covering the shell portion, wherein the coat portion comprises an inorganic phosphor and a target selective binding moiety, and wherein the assay particle is buoyant in aqueous media. An additional type of assay particle includes a core portion encased by a shell portion, and a coat portion covering the shell portion, wherein the shell portion comprises an inorganic phosphor and the coat portion comprises a target selective binding moiety, and wherein the assay particle is buoyant in aqueous media. Also provided are kits and related methods.
1. A method for detecting protein kinase activity, the method comprising:
contacting a sample suspected of containing a protein kinase with:
(i) an assay particle, wherein the assay particle comprises:
a core portion encased by a shell portion; and
a coat portion covering the shell portion,
wherein the shell portion of the assay particle comprises titanium dioxide and a material selected from the group consisting of glass, ceramic, and combinations thereof;
wherein the coat portion comprises titanium dioxide and a target selective binding agent; and
wherein the assay particle is hollow and is buoyant in aqueous media; and
(ii) a protein kinase substrate comprising a moiety capable of emitting radiation under conditions wherein the protein kinase can phosphorylate the substrate to produce a phosphorylated substrate;
wherein binding of the phosphorylated substrate to titanium dioxide produces a light signal; and
determining a protein kinase activity based on a level of detected light signal.
2. The method of claim 1 , wherein the core portion comprises a gas selected from the group consisting of air, nitrogen, oxygen, and combinations thereof.
3. The method of claim 1 , wherein the assay particle has a density of less than 1 g/cm 3 .
4. The method of claim 1 , wherein the shell portion of the assay particle comprises glass.
5. The method of claim 1 , wherein the shell portion of the assay particle comprises ceramic.
6. The method of claim 1 , wherein the core portion comprises air.
7. The method of claim 1 , wherein the core portion comprises nitrogen.
8. The method of claim 1 , wherein the core portion comprises oxygen.
9. The method of claim 1 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
10. The method of claim 1 , wherein the coat portion has a thickness of less than 3000 μm.
11. The method of claim 1 , wherein the coat portion has a thickness of less than 2000 μm.
12. The method of claim 1 , wherein the coat portion has a thickness of less than 1000 μm.
13. The method of claim 1 , wherein the shell portion of the assay particle comprises a target selective binding agent.
14. The method of claim 13 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
15. The method of claim 1 , wherein the shell portion of the assay particle has a thickness of about 1 mm to about 500 mm.
16. The method of claim 1 , wherein the shell portion of the assay particle has a thickness of about 1 mm to about 200 mm.
17. The method of claim 1 , wherein the assay particle comprises an outer layer covering the coat portion.
18. The method of claim 17 , wherein the outer layer comprises a target selective binding agent.
19. The method of claim 18 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
20. The method of claim 1 , wherein the assay particle comprises an encoding element.
21. The method of claim 20 , wherein the encoding element comprises a radio frequency identifier, holographic identifier, fluorophore, or quantum dot.
22. A method for detecting protein kinase activity, the method comprising:
contacting a sample suspected of containing a protein kinase with:
(i) an assay particle, wherein the assay particle comprises:
a core portion encased by a shell portion; and
a coat portion covering the shell portion,
wherein the shell portion of the assay particle comprises a target selective binding agent and a material selected from the group consisting of glass, ceramic, and combinations thereof;
wherein the coat portion comprises titanium dioxide and a target selective binding agent; and
wherein the assay particle is hollow and is buoyant in aqueous media; and
(ii) a protein kinase substrate comprising a moiety capable of emitting radiation under conditions wherein the protein kinase can phosphorylate the substrate to produce a phosphorylated substrate;
wherein binding of the phosphorylated substrate to titanium dioxide produces a light signal; and
determining a protein kinase activity based on a level of detected light signal.
23. The method of claim 22 , wherein the core portion comprises a gas selected from the group consisting of air, nitrogen, oxygen, and combinations thereof.
24. The method of claim 22 , wherein the assay particle has a density of less than 1 g/cm 3 .
25. The method of claim 22 , wherein the shell portion of the assay particle comprises glass.
26. The method of claim 22 , wherein the shell portion of the assay particle comprises ceramic.
27. The method of claim 22 , wherein the core portion comprises air.
28. The method of claim 22 , wherein the core portion comprises nitrogen.
29. The method of claim 22 , wherein the core portion comprises oxygen.
30. The method of claim 22 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
31. The method of claim 22 , wherein the coat portion has a thickness of less than 3000 μm.
32. The method of claim 22 , wherein the coat portion has a thickness of less than 2000 μm.
33. The method of claim 22 , wherein the coat portion has a thickness of less than 1000 μm.
34. The method of claim 22 , wherein the shell portion of the assay particle comprises titanium dioxide.
35. The method of claim 22 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
36. The method of claim 22 , wherein the shell portion of the assay particle has a thickness of about 1 mm to about 500 mm.
37. The method of claim 22 , wherein the shell portion of the assay particle has a thickness of about 1 mm to about 200 mm.
38. The method of claim 22 , wherein the assay particle comprises an outer layer covering the coat portion.
39. The method of claim 38 , wherein the outer layer comprises a target selective binding agent.
40. The method of claim 39 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
41. The method of claim 22 , wherein the assay particle comprises an encoding element.
42. The method of claim 41 , wherein the encoding element comprises a radio frequency identifier, holographic identifier, fluorophore, or quantum dot.
43. A method for detecting protein kinase activity, the method comprising:
contacting a sample suspected of containing a protein kinase with:
(i) an assay particle, wherein the assay particle comprises:
a core portion encased by a shell portion; and
a coat portion covering the shell portion,
wherein the shell portion of the assay particle comprises a material selected from the group consisting of glass, ceramic, and combinations thereof, and wherein the shell portion of the assay particle has a thickness of about 1 mm to about 500 mm;
wherein the coat portion comprises titanium dioxide and a target selective binding agent; and
wherein the assay particle is hollow and is buoyant in aqueous media; and
(ii) a protein kinase substrate comprising a moiety capable of emitting radiation under conditions wherein the protein kinase can phosphorylate the substrate to produce a phosphorylated substrate;
wherein binding of the phosphorylated substrate to titanium dioxide produces a light signal; and
determining a protein kinase activity based on a level of detected light signal.
44. The method of claim 43 , wherein the core portion comprises a gas selected from the group consisting of air, nitrogen, oxygen, and combinations thereof.
45. The method of claim 43 , wherein the assay particle has a density of less than 1 g/cm 3 .
46. The method of claim 43 , wherein the shell portion of the assay particle comprises glass.
47. The method of claim 43 , wherein the shell portion of the assay particle comprises ceramic.
48. The method of claim 43 , wherein the core portion comprises air.
49. The method of claim 43 , wherein the core portion comprises nitrogen.
50. The method of claim 43 , wherein the core portion comprises oxygen.
51. The method of claim 43 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
52. The method of claim 43 , wherein the shell portion of the assay particle comprises titanium dioxide.
53. The method of claim 43 , wherein the shell portion of the assay particle comprises a target selective binding agent.
54. The method of claim 53 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
55. The method of claim 43 , wherein the shell portion of the assay particle has a thickness of about 1 mm to about 200 mm.
56. The method of claim 43 , wherein the assay particle comprises an outer layer covering the coat portion.
57. The method of claim 56 , wherein the outer layer comprises a target selective binding agent.
58. The method of claim 57 , wherein the target selective binding agent is selected from the group consisting of an antibody, aptamer, protein A, protein G, streptavidin, avidin, captavidin, neutravidin, metal chelate, siderophore, lectin, and oligonucleotide.
59. The method of claim 43 , wherein the assay particle comprises an encoding element.
60. The method of claim 59 , wherein the encoding element comprises a radio frequency identifier, holographic identifier, fluorophore, or quantum dot.