Polymer Conjugate Enhanced Bioassays

• US 20080200562A1
• Filed: 05/02/2006
• Published: 08/21/2008
• Est. Priority Date: 05/02/2005
• Status: Active Grant

• Current Assignee: ANP Technologies, Inc.
• Original Assignee: ANP Technologies, Inc.
• Inventors: Pan, Jing, Yin, Ray, Qin, Dujie
• Granted Patent: US 8,563,329 B2
• Time in Patent Office: Days
• US Class Current: 514/772.3
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

Modified branched polymers are combined with bioactive agents which are one member of a binding pair for use in an assay.

Claims

1. A modified branched polymer conjugate which comprises at least one modified branched polymer associated with at least one bioactive agent

2. The conjugate of claim 1, wherein said polymer is symmetric.

3. The conjugate of claim 1, wherein said polymer is asymmetric.

4. The conjugate of claim 1, wherein said bioactive agent is a diagnostic agent, therapeutic agent, or a combination of diagnostic and therapeutic agent.

5. The conjugate of claim 1, wherein said modified branched polymer connects with another bioactive agent through either covalent or non-covalent linkages.

6. The conjugate of claim 1, wherein said modified branched polymer is either a terminal or a chain branched polymer or a combination thereof.

7. The conjugate of claim 1, wherein said modified branched polymer is derived from a dendrimer, a dendrigraft, a star shaped, or a comb-shaped polymer, or a combination thereof.

8. The conjugate of claim 1, wherein said modified branched polymer is derived from a polyethyleneimine or a polypropyleneimine dendrimer.

9. The conjugate of claim 1, wherein said modified branched polymer is derived from a polyethyleneimine or a polypropyleneimine dendrigrafts.

10. The conjugate of claim 3, wherein said asymmetrically branched polymer is a random or regular asymmetrically branched polymer.

11. The conjugate of claim 10, wherein said random asymmetrically branched polymer is polyethyleneimine or a modified polyethyleneimine.

12. The conjugate of claim 10 wherein said random asymmetrically branched polymer is polyoxazoline or modified polyoxazoline.

13. The conjugate of claim 10, wherein said random asymmetrically branched polyoxazoline is poly (2-methyoxazoline) or poly (2-ethyloxazoline).

14. The conjugate of claim 10, wherein said random asymmetrically branched polyoxazoline is modified poly (2-methyoxazoline) or modified poly (2-ethyloxazoline).

15. The conjugate of claim 10, wherein said regular asymmetrically branched polymer is polylysine or a modified polylysine.

16. The conjugate of claim 1, wherein said bioactive agent is a metal.

17. The conjugate of claim 16, wherein said metal is a transition metal, an alkali metal, an alkaline-earth metal, a Lanthanide series element or an Actinide series element.

18. The conjugate of claim 1, wherein said conjugate can be used as a diagnostic agent, a therapeutic agent or both.

19. The conjugate of claim 1, wherein said conjugate can be used in conjunction with a nano and/or microparticle as a medical diagnostic agent, a therapeutic agent or both.

20. The conjugate of claim 19, wherein said nano and/or microparticle is biodegradable or non-biodegradable.

21. The conjugate of claim 1 or 19, wherein said conjugate is used for radiotherapy, immunotherapy, cell therapy, gene therapy or drug delivery.

22. An assay kit comprising the conjugate of claim 1 and a reporter molecule.

23. The kit of claim 22, wherein said bioactive agent is a member of a binding pair.

24. The kit of claim 23, wherein said binding pair comprises an antibody, an antigen binding portion thereof, an antigen or an epitope-containing portion thereof.

25. The kit of claim 22, wherein said reporter molecule comprises a colored, luminescent or fluorescent particulate or moiety, an enzyme, or a combination thereof.

26. The kit of claim 25, wherein said fluorescent or luminescent particulate or moiety comprises quantum dots, nanocrystals, up-converting phosphorescent particles or fluorophore containing latex beads.

27. The kit of claim 25, wherein said colored particulate comprises colloidal metals, comprising gold or silver, colored latex beads or colored dyes.

28. The kit of claim 22, wherein said reporter comprises at least one modified branched polymer.

29. The kit of clam 22, wherein said kit comprises a solid phase comprising said conjugate and said reporter molecule.

30. The kit of claim 29, wherein said solid phase comprises a flat surface or a membrane.

31. The kit of claim 29, wherein said flat surface comprises a silicon wafer, quartz, glass, a metal, or a plastic.

32. The kit of claim 30, wherein said membrane comprises a paper, a plastic membrane, a nylon membrane or nitrocellulose.

33. The kit of claim 29, wherein said solid phase and said conjugate comprise a microarray or a bead array.

34. The kit of claim 22, wherein said reporter molecule yields a product which is detectable by color or by light.

35. The kit of claim 29, wherein said solid phase comprises a dipstick, a lateral flow immunoassay or a microarray.

36. The kit of claim 23, wherein said member of a binding pair is an antibody that binds to a detector that binds to a ligand or analyte in a sample.

37. The kit of claim 23, wherein said member of a binding pair is a molecule that binds biotin, fluorescein, nucleic acid, albumin, a hapten or a combination thereof.

38. The kit of claim 29, wherein said conjugate is bound to said solid phase.

39. The kit of claim 29, wherein said conjugate is reversibly bound to said solid phase.

40. The kit of claim 22, further comprising a detector molecule that binds to a ligand or analyte in a sample.

41. The kit of claim 40, wherein said detector molecule is an antibody or antigen-binding portion thereof.

42. The kit of claim 41, wherein said antibody or antigen binding portion thereof comprises a member of a binding pair.

43. The kit of claim 42, where said member of a binding pair is biotin, fluorescein, nucleic acid, albumin, a hapten or a combination thereof.

44. The kit of claim 29, further comprising a detector molecule that binds to a ligand or analyte in a sample reversibly bound to said solid phase.

45. The kit of claim 44. wherein said detector molecule is an antibody or antigen-binding portion thereof.

46. The kit of claim 45, wherein said antibody or antigen binding portion thereof comprises a member of a binding pair.

47. The kit of claim 46, wherein said member of a binding pair is biotin fluorescein, nucleic acid, albumin, a hapten or a combination thereof.

48. A method for detecting a target molecule, comprising, obtaining a sample suspected of containing said target molecule, exposing said sample to the conjugate of claim 1 and a detector molecule that specifically binds said target molecule, and determining whether complexes of said conjugate, detector molecule and target molecule are formed.

49. The method of claim 48, wherein said determining step comprises a reporter molecule.

50. The method of claim 49, wherein presence of said reporter molecule is determined visually.

51. The method of claim 49, wherein presence of said reporter molecule is detected with a mechanical means.

52. The method of claim 51, wherein said mechanical means comprises a detecting means.

53. The method of claim 51, wherein said mechanical means comprises a sensing means that digitizes data and a data storage means.

54. The method of claim 51, wherein said mechanical means comprises a display means.

55. The method of claim 51, wherein said mechanical means is a hand held device.

56. The method of claim 51, further comprising a data communication means.

57. The method of claim 51, wherein said mechanical means is portable.

58. The method of claim 51, wherein said mechanical means comprises a detection signal quantifying means.

59. The method of claim 56, wherein said communication means is a wireless means.

60. The kit of claim 22, further comprising a housing.

61. The method of claim 48, wherein said target molecule is immobilized on a surface prior to reacting with said detector molecule.

62. The method of claim 61, wherein said immobilizing comprises a binding molecule which specifically binds said target affixed to said surface.