Method and apparatus for molecular analysis in small sample volumes

• US 7,344,832 B2
• Filed: 12/30/2003
• Issued: 03/18/2008
• Est. Priority Date: 01/02/2003
• Status: Expired due to Fees

• Current Assignee: BioForce Nanosciences, Inc.
• Original Assignee: BioForce Nanosciences, Inc.
• Inventors: Mosher, Curtis, Huff, Janice, Henderson, Eric
• Primary Examiner(s): Lam; Ann Y.
• Application Number:
  US10/541,013
• Publication Number:
  US 20060035234A1
• Time in Patent Office: 1,540 Days
• Field of Search: 435/6, 435/4, 435/7.1, 435/7.92, 435/283.1, 435/287.1, 435/287.2, 436/518, 422/50, 422/58
• US Class Current: 435/6.19
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 35/00 Methods or apparatus for measurement or analysis of nanostructures
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01Q 60/42 Functionalisation

Abstract

The interrogation of extremely small sample volumes can be accomplished with the present invention. Provided are probes having disposed thereon a plurality of domains forming an array, which is suitably a nanoarray. Also provided are methods of detecting molecules and molecular interaction events, retrieving and analyzing analytes, and delivering substances to cells or tissues using probes of the invention.

Claims

1. An apparatus for analyzing a sample comprising a probe, the probe comprising a pointed member, the pointed member having a plurality of domains disposed thereon, wherein the domains comprise one or more biomolecules and wherein the domains form an array.

2. The apparatus of claim 1, wherein the array is a nanoarray.

3. The apparatus of claim 1, wherein the one or more biomolecules are selected from the group consisting of drugs, drug candidates, chemical groups, lipids, DNA, RNA, proteins, peptide species, carbohydrates, and any combination thereof.

4. The apparatus of claim 1, further comprising nanosensors operably connected to one or more of the domains.

5. The apparatus of claim 1, wherein the probe is a dual element probe.

6. The apparatus of claim 1, wherein the probe is a multielement probe.

7. The apparatus of claim 1, wherein the probe is sized to interrogate a sample comprising a volume of about 50 femtoliters to about 10 microliters.

8. The apparatus of claim 1, the apparatus comprising at least one microdisrupter disposed on the probe.

9. The apparatus of claim 8, wherein the microdisrupter comprises the pointed member.

10. The apparatus of claim 1, wherein the probe further comprises at least one hydrophobic region.

11. The apparatus of claim 1, further comprising a molecular detection device operably connected to the probe.

12. The apparatus of claim 11, wherein the molecular detection device is a scanning tunneling microscope, atomic force microscope, mass spectrometer, fluorescence microscope, flow cytometer, Raman spectrometer, Infra-red spectrometer, UV spectrometer, electronic system, electrochemical system, optical system, magnetic and electromagnetic system, or mass measuring system.

13. A method of detecting a molecular interaction event comprising:

contacting a sample with the probe of claim 1;

providing an incubation period;

washing unbound molecules from the domains; and

detecting the molecular interaction event.

14. The method of claim 13 wherein the sample comprises at least one cell.

15. The method of claim 13 wherein the sample comprises at least one cell lysate.

16. A method of detecting one or more molecules in a sample comprising:

contacting the sample with the probe of claim 4; and

detecting binding of one or more molecules to one or more of the domains.

17. The apparatus of claim 1, wherein the domains are spatially arranged in known locations.

18. The apparatus of claim 1, wherein the probe is sized to interrogate a single cell.

19. The apparatus of claim 1, wherein the probe is sized to interrogate a lysate of a single cell.

20. The apparatus of claim 1, wherein the probe is sized to interrogate a sub-cellular species of a cell.

21. The apparatus of claim 20, wherein the sub-cellular species is selected from the group consisting of a Golgi complex, a mitochondria, a lysosome, an endoplasmic reticulum, a lipid raft, and a cytoskeletal system.

22. The apparatus of claim 1, wherein the pointed member is sized to be inserted into a cell.

23. The apparatus of claim 1, wherein the pointed member comprises an anti-wicking feature.

24. The apparatus of claim 23, wherein the anti-wicking feature comprises a hydrophobic domain.

25. The apparatus of claim 1, wherein at least one domain has a substance reversibly attached thereto.

26. The apparatus of claim 25, wherein the at least one domain is reversibly attached by a tether, the tether comprising a protease substrate, a photolyzable tether, a chemically reactive tether, an ionically reactive tether, or a thermally sensitive tether.

27. A method of delivering at least one substance to a cell, comprising:

passing the pointed member of the probe of claim 25 through the membrane of the cell into the intracellular space; and

releasing the substance into the intracellular space.

28. A method of analyzing one or more analytes in a cell, comprising:

passing the pointed member of the probe of claim 1 through the membrane of the cell into the intracellular space; and

detecting the binding of the analyte to the domains of the array.

29. The method of claim 28, wherein the array is a nanoarray.

30. A method of retrieving an analyte from a cell, comprising:

passing the pointed member of the probe of claim 1 through the membrane of the cell into the intracellular space, wherein the probe has at least one domain capable of binding to the analyte; and

retrieving the analyte from the domain.

31. A method of detecting an in situ molecular interaction event comprising:

contacting a sample with the pointed member of the probe of claim 1; and

detecting the molecular interaction event.

32. The apparatus of claim 3 wherein the one or more biomolecule is a drug.

33. The apparatus of claim 3 wherein the one or more biomolecule is a drug candidate.

34. The apparatus of claim 3 wherein the one or more biomolecule is a chemical group.

35. The apparatus of claim 3 wherein the one or more biomolecule is a lipid.

36. The apparatus of claim 3 wherein the one or more biomolecule is DNA.

37. The apparatus of claim 3 wherein the one or more biomolecule is RNA.

38. The apparatus of claim 3 wherein the one or more biomolecule is a protein.

39. The apparatus of claim 3 wherein the one or more biomolecule is a peptide species.

40. The apparatus of claim 3 wherein the one or more biomolecule is a carbohydrate.

41. An apparatus for analyzing a sample comprising a probe, the probe comprising a pointed member, the pointed member having a plurality of domains disposed thereon, wherein the domains form an array and wherein the probe is sized to interrogate a single cell.

42. An apparatus for analyzing a sample comprising a probe, the probe comprising a pointed member, the pointed member having a plurality of domains disposed thereon, wherein the domains form an array and wherein the pointed member is sized to be inserted into a cell.

43. An apparatus for analyzing a sample comprising a probe, the probe comprising a pointed member, the pointed member having a plurality of domains disposed thereon, wherein the domains form an array and wherein at least one domain has a substance reversibly attached thereto reversibly attached by a tether, the tether comprising a protease substrate, a photolyzable tether, a chemically reactive tether, an ionically reactive tether, or a thermally sensitive tether.