Method and device for extracting an analyte

• US 20050255604A1
• Filed: 08/17/2004
• Published: 11/17/2005
• Est. Priority Date: 05/12/2004
• Status: Abandoned Application

• Current Assignee: PhyNexus, Inc.
• Original Assignee: PhyNexus, Inc.
• Inventors: Burge, Allen, Gjerde, Douglas T., Hanna, Christopher P.
• Application Number:
  US10/920,901
• Publication Number:
  US 20050255604A1
• Time in Patent Office: Days
• US Class Current: 436/101
• CPC Class Codes: B01D 15/325 Reversed phase
• CPC Class Codes: B01D 15/327 with hydrophobic interaction
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01J 20/285 based on polymers
• CPC Class Codes: B01J 20/291 Gel sorbents
• CPC Class Codes: B01J 2220/64 In a syringe, pipette, e.g. tip or in a tube, e.g. test-tube or u-shape tube
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 3/0275 Interchangeable or disposable dispensing tips
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 2030/009 Extraction
• CPC Class Codes: G01N 2030/525 surface properties, e.g. porosity
• CPC Class Codes: G01N 2035/103 using disposable tips
• CPC Class Codes: G01N 2035/1039 Micropipettes, e.g. microcapillary tubes
• CPC Class Codes: G01N 2035/1053 for separating part of the liquid, e.g. filters, extraction phase
• CPC Class Codes: G01N 30/6004 end pieces
• CPC Class Codes: G01N 35/10 Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: Y10T 436/153333 Halogen containing
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The invention provides extraction columns for the purification of an analyte (e.g., a biological macromolecule, such as a peptide, protein or nucleic acid) from a sample solution, as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the column between two frits. In some embodiments, the extraction columns employ modified pipette tips as column bodies.

Claims

1. A separation column comprising:

a. an outer column body having a longitudinal axis, a first open channel, a first open upper end, a first open lower end, and a first frit bonded to and extending across the first open lower end, wherein the section of the first open channel between the first frit and the first open upper end defines an outer column body volume;

b. an inner column body having a second open channel, a second open upper end, a second open lower end, and a second frit bonded to and extending across the second open lower end, wherein at least some portion of the inner column body is sealingly positioned within the first open channel between the first frit and first open upper end, said portion including the second open lower end and second frit;

c. a media chamber defined by the first frit, the second frit and the surface of the first open channel, wherein said media chamber has a volume that is less than the outer column body volume; and

d. a bed of media positioned within said media chamber.

2. The separation column of claim 1, wherein the inner column body is stably affixed to the outer column by a frictional engagement with the surface of the first open channel.

3. The separation column of claim 2, wherein the entire inner column body is disposed within the first open channel.

4. The separation column of claim 3, wherein the first open upper end is adapted for operable attachment to a pump.

5. The separation column of claim 4, wherein the outer column body is a pipette tip and the pump is a pipettor.

6. The separation column of claim 5, wherein the outer diameter of the inner column body tapers towards the second open lower end, and wherein the first open channel is tapered in the region where the inner column body frictionally engages the first open channel, the tapers of the inner column body and first open channel being complementary to one another.

7. The separation column of claim 2, wherein the second open upper end is adapted for operable attachment to a pump.

8. The separation column of claim 7, wherein some portion of the inner column body is not disposed within the first open channel, and said portion includes the second open upper end.

9. The separation column of claim 7, wherein the inner column body is a pipette tip and the pump is a pipettor.

10. The separation column of claim 7, wherein the outer diameter of the inner column body tapers towards the second open lower end, and wherein the first open channel is tapered in the region where the inner column body frictionally engages the first open channel, the tapers of the inner column body and first open channel being complementary to one another.

11. The separation column of claim 1, wherein the bed of media comprises a packed bed of gel-based extraction beads.

12. The separation column of claim 1, wherein said first and second frits are membrane frits.

13. The separation column of claim 5, wherein said first and second membrane frits are bonded to said outer column body and said ring, respectively, by gluing or welding.

14. The separation column of claim 5, wherein said first and second membrane frits comprises a polyester mesh.

15. The separation column of claim 5, wherein said outer column body comprises polycarbonate.

16. The separation column of claim 1, wherein the inner column body is disposed so that the second frit is substantially perpendicular to the longitudinal axis of the outer column body.

17. A method for manufacturing a separation column comprising the steps of:

a. providing an outer column body having a longitudinal axis, a first open channel, a first open upper end, a first open lower end, and a first frit bonded to and extending across the first open lower end, wherein the section of the first open channel between the first frit and the first open upper end defines an outer column body volume, and wherein a quantity of media is positioned within the first open channel; and

b. sealingly positioning at least some portion of an inner column body within the first open channel between the first frit and first open upper end, wherein the inner column body has a second open channel, a second open upper end, a second open lower end, and a second frit bonded to and extending across the second open lower end, said portion including the second open lower end and second frit such that the first frit, the second frit and the surface of the first open channel define a media chamber containing the quantity of media, and wherein the volume of the media chamber is less than the outer column body volume.

18. A separation column comprising:

a. an outer column body having a longitudinal axis, an open channel, a first open upper end adapted for operable attachment to a pump, a first open lower end for the uptake and dispensing of fluid, the open lower end having a first internal diameter, and a first frit bonded to and extending across the first open lower end, wherein the section of the first open channel adjacent to the open lower end is tapered, the inner diameter of the tapered section decreasing from a second internal diameter to the first internal diameter of the first open lower end;

b. a ring having an outer diameter that is greater than the first internal diameter and less than the second internal diameter, having a second frit bonded to and extending across said ring, said ring disposed within said tapered section, the ring sealingly engaging the surface of the open channel; and

c. a bed of media disposed within a media chamber defined by the first frit, the second frit and the surface of the open channel.

19. The separation column of claim 18, wherein said ring comprises a frustoconical member having a central through passageway and open opposite first and second ends, wherein the outer diameter of the frustoconical member tapers from a first outer diameter of the first end that is greater than the second internal diameter of the outer column body, to a second outer diameter of the second end that is greater than the first internal diameter of the open lower end of the column body and less than the second internal diameter of the column body, said second frit bonded to and extending across the second end of said frustoconical member.

20. The separation column of claim 18, wherein the ring is frictionally engaged with the surface of the open channel.

21. The separation column of claim 18, wherein bed of media comprises a packed bed of gel-based extraction beads.

22. The separation column of claim 18, wherein said first and second frits are membrane frits.

23. The separation column of claim 22, wherein said first and second membrane frits are bonded to said outer column body and said ring, respectively, by gluing or welding.

24. The separation column of claim 22, wherein said first and second membrane frits comprises a polyester mesh.

25. The separation column of claim 22, wherein said outer column body comprises a pipette tip.

26. The separation column of claim 18, wherein said ring is disposed so that the second frit is substantially perpendicular to the longitudinal axis of the outer column body.

27. A method of manufacturing a separation column comprising:

a. providing an outer column body having a longitudinal axis, an open channel, a first open upper end adapted for operable attachment to a pump, a first open lower end for the uptake and dispensing of fluid, the open lower end having a first internal diameter, and a first frit bonded to and extending across the first open lower end, wherein the section of the first open channel adjacent to the open lower end is tapered, the inner diameter of the tapered section decreasing from a second internal diameter to the first internal diameter of the first open lower end, and wherein a quantity of media is positioned within the open channel; and

b. positioning a ring within said tapered section such that the ring sealingly engages the surface of the open channel, wherein the ring has an outer diameter that is greater than the first internal diameter and less than the second internal diameter, and a second frit bonded to and extending across the ring.