Open channel solid phase extraction systems and methods

• US 20040224362A1
• Filed: 03/04/2004
• Published: 11/11/2004
• Est. Priority Date: 06/10/2002
• Status: Active Grant

• Current Assignee: PhyNexus, Inc.
• Original Assignee: PhyNexus, Inc.
• Inventors: Gjerde, Douglas T., Hanna, Christopher T.
• Granted Patent: US 7,122,640 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 2030/009 Extraction
• CPC Class Codes: G01N 2030/062 extracting sample from raw material

Abstract

The invention provides, inter alia, methods of extracting an analyte from a solution comprising the steps of: passing a solution containing an analyte through an extraction channel having a solid phase extraction surface, whereby analyte adsorbs to the extraction surface of said extraction channel; purging bulk liquid from said extraction channel; and eluting the analyte by passing a desorption solvent through the channel. The invention further provides reagents, columns and instrumentation related to this and other methods.

Claims

1. A method of extracting an analyte from a solution comprising the steps of:

a. passing a solution containing an analyte through an extraction channel having a solid phase extraction surface, whereby analyte adsorbs to the extraction surface of said extraction channel;

b. purging bulk liquid from said extraction channel; and

c. eluting the analyte by passing a desorption solvent through the channel.

2. The method of claim 1, wherein as a result of the purge step the extraction channel is at least 50% free of bulk liquid.

3. The method of claim 2, wherein as a result of the purge step the extraction channel is at least 90% free of bulk liquid.

4. The method of claim 3, wherein as a result of the purge step the extraction channel is substantially free of bulk liquid.

5. The method of claim 1, wherein said extraction surface remains substantially solvated during step (b).

6. The method of claim 1, wherein said extraction channel is purged by blowing a gas through the channel.

7. The method of claim 6, wherein the gas is nitrogen or helium.

8. The method of claim 1, wherein a wash solution is passed through the channel between steps (a) and (b).

9. The method of claim 1, wherein said extraction channel comprises a capillary.

10. The method of claim 9, wherein said capillary is a fused silica capillary.

11. The method of claim 1, wherein said analyte is a biomolecule.

12. The method of claim 1, wherein said biomolecule is a protein.

13. The method of claim 12, wherein the eluted protein retains its native function.

14. The method of claim 1, wherein said biomolecule is a multi-protein complex.

15. The method of claim 1, wherein said analyte is eluted with a tube enrichment factor of at least one.

16. The method of claim 1, wherein the analyte containing solution is passed through the extraction channel at least twice.

17. The method of claim 1, wherein the desorption solvent is passed through the extraction channel at least twice.

18. The method of claim 1, wherein the extraction surface comprises an affinity binding agent.

19. The method of claim 18, wherein the affinity binding agents is:

a) a chelated metal having a binding affinity for a biomolecule analyte;

b) a protein having a binding affinity for a protein analyte;

c) an organic molecule or group having a binding affinity for a protein analyte;

d) a sugar having a binding affinity for a protein analyte;

e) nucleic acid having a binding affinity for a protein analyte;

f) a nucleic acid or a sequence of nucleic acids having a binding affinity for a nucleic acid analyte;

or

g) a small molecule binding agent having a binding affinity for a small molecule analyte.

20. The method of claim 1, wherein the solid phase extraction surface is a 3-dimensional solid phase extraction surface.

21. The method of claim 20, wherein the solid-phase extraction surface comprises a polymer.

22. The method of claim 21, wherein the polymer is a polysaccharide.

23. The method of claim 1, wherein the extraction surface is at least partially desolvated during step (b).

40. The method of claim 1, wherein the volume of desorption solvent is in the range of 10 pL to 500 μ

L.

24. An extraction channel comprising:

a. a channel;

b. a solid phase extraction surface bound to the inner surface of said channel; and

c. an analyte reversibly bound to said extraction surface, wherein the extraction channel is at least 50% free of bulk liquid.

25. The extraction channel of claim 24, wherein said analyte is a biomolecule.

26. The extraction channel of claim 25, wherein said biomolecule is a protein.

27. The extraction channel of claim 25, wherein the extraction channel is at least 90% free of bulk liquid.

28. The extraction channel of claim 27, wherein the extraction channel is substantially free of bulk liquid.

29. The extraction channel of claim 24, wherein the extraction surface is substantially solvated.

30. The extraction channel of claim 24, wherein said channel comprises a capillary.

31. The extraction channel of claim 30, wherein said capillary is a fused silica capillary.

32. The extraction channel of claim 26, wherein said protein is not denatured.

33. The extraction channel of claim 25, wherein said biomolecule is a multi-protein complex.

34. The extraction channel of claim 25, wherein the extraction surface comprises an affinity binding agent, and said analyte is reversibly bound to said affinity binding agent.

35. The extraction channel of claim 34, wherein the affinity binding agents is:

a) a chelated metal having a binding affinity for a biomolecule analyte;

b) a protein having a binding affinity for a protein analyte;

c) an organic molecule or group having a binding affinity for a protein analyte;

d) a sugar having a binding affinity for a protein analyte;

e) nucleic acid having a binding affinity for a protein analyte;

f) a nucleic acid or a sequence of nucleic acids having a binding affinity for a nucleic acid analyte;

or

g) a small molecule binding agent having a binding affinity for a small molecule analyte.

36. The extraction channel of claim 24, wherein the solid phase extraction surface is a 3-dimensional solid phase extraction surface.

37. The extraction channel of claim 36, wherein the solid-phase extraction surface comprises a polymer.

38. The extraction channel of claim 37, wherein the polymer is a polysaccharide.

39. The extraction channel of claim 24, wherein said extraction surface is at least partially desolvated.