Method and apparatus for molecular analysis in small sample volumes

• US 20060035234A1
• Filed: 12/30/2003
• Published: 02/16/2006
• Est. Priority Date: 01/02/2003
• Status: Active Grant

• Current Assignee: Curtis Mosher, Eric Henderson, Janice Huff
• Original Assignee: Curtis Mosher, Eric Henderson, Janice Huff
• Inventors: Henderson, Eric, Mosher, Curtis, Huff, Janice
• Granted Patent: US 7,344,832 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 having a plurality of domains disposed thereon, 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 domains comprise one or more biomolecules 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 comprises a microcantilever.

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

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

8. The apparatus of claim 1, wherein the sample comprises a volume of about 50 femtoliters to about 10 microliters.

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

10. The apparatus of claim 9, wherein at least one microdisrupter comprises a tip or pointed member.

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

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

13. The apparatus of claim 12, 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.

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

contacting a sample with a probe having a plurality of domains disposed in an array;

providing an incubation period;

washing unbound molecules from the domains; and

detecting the molecular interaction event.

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

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

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

contacting the sample a probe having a plurality of domains disposed thereon, wherein the domains form an array, and wherein the domains are operably connected to one or more nanosensors; and

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

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

disrupting a cell with a microdisrupter disposed on a probe, wherein the probe has a plurality of domains disposed thereon, and wherein the domains form a nanoarray;

passing the nanoarray through the membrane of the cell such that the nanoarray contacts intracellular space; and

detecting the binding of one or more analytes to the nanoarray.

19. The method of claim 18, further comprising passing the probe through the nuclear membrane such that the nanoarray contacts intranuclear space.

20. The method of claim 18, further comprising inserting the probe into a sub-cellular species.

21. The method 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. A method of retrieving at least one analyte from a sample comprising:

contacting the sample with a probe having a plurality of domains disposed thereon, wherein the domains form an array; and

retrieving at least one analyte from the molecular domains.

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

reversibly attaching at least one substance to a probe having a plurality of domains disposed thereon, wherein the domains form an array;

passing the probe through the membrane of the cell into the intracellular space; and

releasing at least one substance into the intracellular space.

24. The method of claim 23 wherein reversibly attaching at least one substance to a probe comprises contacting the substance to the domains such that a binding event occurs.

25. The method of claim 23 wherein at least one substance is DNA, RNA, a peptide species, a chemical, a drug or a reactive species.

26. The method of claim 23 wherein reversibly attaching comprises tethering at least one substance to at least one domain with a protease substrate, a photolyzable tether, a chemically reactive tether, an ionically reactive tether or a thermally sensitive tether.

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

contacting a sample with a probe having a plurality of domains disposed in an array;

providing an incubation period; and

detecting the molecular interaction event.