Asymmetrically branched polymer conjugates and microarray assays

• US 20110065090A1
• Filed: 07/12/2010
• Published: 03/17/2011
• Est. Priority Date: 11/21/2003
• Status: Active Grant

• Current Assignee: ANP Technologies, Inc.
• Original Assignee: ANP Technologies, Inc.
• Inventors: Pan, Jing, Yin, Ray, Qin, Dujie
• Granted Patent: US 8,501,399 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• 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

A method of detection comprising a conjugate of a randomly and asymmetrically branched dendritic polymer.

Claims

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

2. The method for detecting a target molecule of claim 1, further comprising,

exposing said sample suspected of containing said target molecule to:

a. a first member of a first binding pair, wherein said first member of said first binding pair binds said target molecule, and wherein said first member further comprises a first member of a second binding pair, and then to

b. said conjugate, wherein said conjugate comprises

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

(b) a second member of said second binding pair;

wherein said complexes are formed on a solid phase comprising a first member of a third binding pair, wherein said first member of said third binding pair binds said target molecule.

3. The method of claim 2, wherein said determining step comprises a reporter.

4. The method of claim 3, wherein presence of said reporter is determined visually.

5. The method of claim 3, wherein presence of said reporter molecule is detected with a mechanical device.

6. The method of claim 5, wherein said mechanical device comprises a detecting device.

7. The method of claim 5, wherein said mechanical device comprises a sensing device that digitizes data and a data storage device.

8. The method of claim 5, wherein said mechanical device comprises a display device.

9. The method of claim 5, wherein said mechanical device is a hand held device.

10. The method of claim 5, further comprising a data communication device.

11. The method of claim 5, wherein said mechanical device is portable.

12. The method of claim 5, wherein said mechanical device comprises a detection signal quantifying device.

13. The method of claim 10, wherein said communication device is wireless.

14. The method of claim 2, wherein said randomly branched dendritic polymer comprises a modified polyethyleneimine.

15. The method of claim 2, wherein said randomly branched dendritic polymer comprises a modified polyoxazoline.

16. The method of claim 15, wherein said modified polyoxazoline comprises a modified poly (2-methyoxazoline) or a modified poly (2-ethyloxazoline).

17. The method of claim 2, wherein said randomly branched dendritic polymer comprises a modified polylysine.

18. The method of claim 2, wherein said first member of said first, second or third binding pair comprises a polynucleotide, an antibody, an antigen-binding portion thereof, an antigen or an epitope-containing portion thereof.

19. The method of claim 3, wherein said reporter comprises a colored, luminescent or fluorescent particulate or moiety, an enzyme, or a combination thereof.

20. The method of claim 19, wherein said fluorescent or luminescent particulate or moiety comprises quantum dots, nanocrystals, up-converting phosphorescent particles, a Lanthanide metal or fluorophore containing latex beads.

21. The method of claim 19, wherein said colored particulate or moiety comprises colloidal metals, comprising gold or silver, colored latex beads or colored dyes.

22. The method of claim 3, wherein said reporter yields a product which is detectable by color or by light.

23. The method of claim 2, wherein said first member of said first binding pair binds to a biological polymer, a hormone, a neurotransmitter, a pathogen, a toxin, or a drug.

24. The method of claim 23, wherein said pathogen comprises a bacterium, a spore, a parasite or a virus.

25. The method of claim 23, wherein said biological polymer comprises a polypeptide, a polysaccharide or a polynucleotide.

26. The method of claim 25, wherein said polypeptide comprises an enzyme.

27. The method of claim 2, wherein said target molecule comprises a polyethylene glycol.

28. The method of claim 2, wherein said second member of said second binding pair comprises an avidin, an antibody which binds a drug, or both, wherein said drug comprises a digoxin.

29. The method of claim 2, wherein said second member of said second binding pair comprises a biotin, an antibody which binds a drug, or both, wherein said drug comprises a digoxin.

30. The method of claim 2, wherein said second member of said second binding pair comprises streptavidin.

31. The method of claim 3, wherein said reporter comprises colloidal gold.

32. The method of claim 2, wherein said solid phase comprises a membrane.

33. The method of claim 32, wherein said membrane comprises nitrocellulose.

34. An method for determining presence of a first member of a binding pair in a sample, comprising, exposing said sample and then a conjugate comprising:

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

(b) a bioactive molecule;

to a solid phase comprising a second member of said binding pair, wherein said bioactive molecule is a first member of said binding pair, and wherein said bioactive molecule of said conjugate and said first member of said binding pair of said sample compete for binding to said second member.

35. The method of claim 34, wherein said conjugate comprises a reporter.

36. The method of claim 34, wherein said randomly branched dendritic polymer comprises a modified polyethyleneimine.

37. The method of claim 34, wherein said randomly branched dendritic polymer comprises a modified polyoxazoline.

38. The method of claim 37, wherein said modified polyoxazoline comprises a modified poly (2-methyoxazoline) or a modified poly (2-ethyloxazoline).

39. The method of claim 34, wherein said randomly branched dendritic polymer comprises a modified polylysine.