Asymmetrically branched polymer conjugates and microarray assays

• US 7,754,500 B2
• Filed: 11/22/2004
• Issued: 07/13/2010
• Est. Priority Date: 11/21/2003
• Status: Active Grant

• Current Assignee: ANP Technologies, Inc.
• Original Assignee: ANP Technologies, Inc.
• Inventors: Qin, Dujie, Pan, Jing, Yin, Ray
• Primary Examiner(s): Cano; Milton I
• Assistant Examiner(s): Mesh; Gennadiy
• Application Number:
  US10/993,474
• Publication Number:
  US 20060041058A1
• Time in Patent Office: 2,059 Days
• Field of Search: 424/78.17, 435/975, 436/525, 436/528, 436/529, 436/546
• US Class Current: 436/525
• CPC Class Codes: A61K 47/6949 inclusion complexes, e.g. clathrates, cavitates or fullerenes
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• 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 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10T 436/11 Automated chemical analysis

Abstract

Asymmetrically branched polymers are combined with bioactive agents for a variety of purposes including drug delivery and conjugation to one member of a binding pair for use in an assay.

Claims

1. A kit comprising:

(1) a reagent comprising a randomly branched dendritic polymer conjugate which comprises;

(a) at least one modified randomly branched dendritic polymer without a core comprising chain branches, terminal branches or both, wherein said randomly branched dendritic polymer is modified to comprise a functional group for branch formation, said randomly branched dendritic polymer comprising random, asymmetric branches and random, asymmetric branch junctions; and

covalently conjugated thereto (b) at least one bioactive agent; and

(2) a reporter.

2. The kit of claim 1, wherein said randomly branched dendritic polymer connects with another bioactive agent.

3. The kit of claim 1, wherein said randomly branched dendritic polymer comprises a modified polyethyleneimine.

4. The kit of claim 1, wherein said randomly branched dendritic polymer comprises a modified polyoxazoline.

5. The kit of claim 4, wherein said modified polyoxazoline comprises a modified poly(2-methyoxazoline) or a modified poly(2-ethyloxazoline).

6. The kit of claim 1, wherein said randomly branched dendritic polymer comprises a modified polylysine.

7. The kit of claim 1, wherein said bioactive agent comprises a metal.

8. The kit of claim 7, wherein said metal comprises a transition metal, an alkali metal, an alkaline-earth metal, a Lanthanide series element or an Actinide series element.

9. The kit of claim 1, wherein said bioactive agent comprises a member of a binding pair.

10. The kit of claim 9, wherein said binding pair comprises a polynucleotide, an antibody, an antigen-binding portion thereof, an antigen or an epitope-containing portion thereof.

11. The kit of claim 1, wherein said reporter comprises a colored, luminescent or fluorescent particulate or moiety, an enzyme, or a combination thereof.

12. The kit of claim 11, wherein said fluorescent or luminescent particulate or moiety comprises quantum dots, nanocrystals, up-converting phosphorescent particles, a Lanthanide metal or fluorophore containing latex beads.

13. The kit of claim 11, wherein said colored particulate or moiety comprises colloidal metals, comprising gold or silver, colored latex beads or colored dyes.

14. The kit of claim 1, wherein said kit comprises a solid phase comprising said reagent and said reporter.

15. The kit of claim 14, wherein said solid phase comprises a flat surface or a membrane.

16. The kit of claim 15, wherein said flat surface comprises a silicon wafer, quartz, glass, a metal, or a plastic.

17. The kit of claim 15, wherein said membrane comprises a paper, a plastic membrane, a nylon membrane or nitrocellulose.

18. The kit of claim 14, wherein said solid phase and said reagent comprise a microarray or a bead array.

19. The kit of claim 1, wherein said reporter yields a product which is detectable by color or by light.

20. The kit of claim 1, wherein said bioactive agent binds to a biological polymer, a hormone, a neurotransmitter, a pathogen, a toxin, or a drug.

21. The kit of claim 20, wherein said pathogen comprises a bacterium, a spore, a parasite or a virus.

22. The kit of claim 20, wherein said biological polymer comprises a polypeptide, a polysaccharide or a polynucleotide.

23. The kit of claim 22, wherein said polypeptide comprises an enzyme.

24. The kit of claim 22, wherein said polypeptide comprises an antibody, an antigen-binding portion thereof, an antigen or an epitope-containing portion thereof.

25. The kit of claim 14, wherein said solid phase comprises a dipstick, a microtiter plate, a lateral flow immunoassay or a microarray.

26. The kit of claim 1, wherein said bioactive agent comprises a drug.

27. The kit of claim 26, wherein said drug comprises an anesthetic, antibiotic, antifungal, antiviral, analgesic, antihypertensive, anti-inflammatory, antidote, antihistamine, chemotherapeutic agent, antidepressant, depressant, stimulant, tranquilizer, urinary anti-infective, vasoconstrictor, vitamin, cardioactive drug, immunosuppressive, or a nutritional supplement.

28. The kit of claim 26, wherein said drug comprises a biological polymer.

29. The kit of claim 26, wherein said drug comprises a hormone.

30. The kit of claim 28, wherein said biological polymer comprises a polypeptide or a polynucleotide.

31. The kit of claim 30, wherein said polynucleotide comprises a DNA or an RNA.

32. The kit of claim 30, wherein said polypeptide comprises ligand or antigen binding.

33. The kit of claim 32, wherein said antigen binding polypeptide comprises an immunoglobulin or antigen-binding portion thereof.

34. The kit of claim 31, wherein said RNA comprises an RNAi.

35. The kit of claim 1, wherein said polymer comprises a modified polyethylene glycol.

36. The kit of claim 12, wherein said Lanthanide metal is Europium.

37. The kit of claim 14, wherein said solid phase comprises a holder.

38. The kit of claim 37, further comprising identifying information.

39. The kit of claim 38, wherein said identifying information comprises a bar code.

40. The kit of claim 1, wherein said bioactive agent comprises an avidin, an antibody which binds a drug, or both, wherein said drug comprises a digoxin.

41. The kit of claim 1, wherein said bioactive agent comprises a biotin, an antibody which binds a drug, or both, wherein said drug comprises a digoxin.

42. The kit of claim 1, wherein said bioactive agent comprises streptavidin.

43. The kit of claim 1, wherein said bioactive agent comprises colloidal gold.

44. The kit of claim 1, further comprising a a device for detecting said reporter or a product of said reporter.

45. The kit of claim 44, wherein said device is hand-held or portable.

46. The kit of claim 44, wherein said device is a scintillation counter, a Geiger counter, a colorimeter, a refractometer, a reflectometer, a scanner, a charge coupled device or a complementary metal oxide semiconductor device.