IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
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출원번호 |
US-0837141
(2010-07-15)
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등록번호 |
US-8148168
(2012-04-03)
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발명자
/ 주소 |
- Gjerde, Douglas T.
- Burge, Allen
- Jones, Ronald
- Abel, Mark
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 |
피인용 횟수 :
13 인용 특허 :
15 |
초록
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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
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, often between two frits. In some embodiments, the extraction columns employ modified pipette tips as column bodies. In some embodiments, the extraction columns are comprised of frits having a low pore. In some embodiments, the frits of the extraction columns have a pore volume of less than one microliter or less than 10% of the interstitial volume of the bed of extraction media.
대표청구항
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1. A pipette tip extraction column comprising: i) a column body having an open upper end, an open lower end, and an open channel between the upper and lower ends of the column body, wherein the column body is comprised of a modified pipette tip;ii) a bottom frit extending across the open channel, sa
1. A pipette tip extraction column comprising: i) a column body having an open upper end, an open lower end, and an open channel between the upper and lower ends of the column body, wherein the column body is comprised of a modified pipette tip;ii) a bottom frit extending across the open channel, said bottom frit having a pore volume of less than one microliter; andiii) a bed of resin media positioned inside the open channel and in contact with the bottom frit. 2. The pipette tip extraction column of claim 1, wherein the bottom frit comprises a hydrophilic membrane screen. 3. The pipette tip extraction column of claim 1, wherein the bottom frit is located at the open lower end of the column body. 4. The pipette tip extraction column of claim 3, wherein the open lower end of the extraction column is in contact with the sample solution, the wash solution or the desorption solution. 5. The pipette tip extraction column of claim 1, wherein the upper end of the column body is operatively attached to a pump for aspirating and discharging fluid through the lower end of the column body. 6. The pipette tip extraction column of claim 5, wherein the pump is a pipettor or a syringe. 7. The pipette tip extraction column of claim 1, wherein the resin media comprises an affinity binding group having an affinity for a biological molecule of interest. 8. The pipette tip extraction column of claim 7, wherein the affinity binding group is selected from the group consisting of Protein A, Protein G, Protein L and an immobilized metal. 9. The pipette tip extraction column of claim 7, wherein the biological molecule is an active protein. 10. The pipette tip extraction column of claim 1, wherein the bed of resin media has a volume of between 0.1 ul and 1000 ul. 11. The pipette tip extraction column of claim 10, wherein the bed of resin media is comprised of silica. 12. The pipette tip extraction column of claim 1, further comprising a top frit extending across the open channel between the bed of resin media and the open upper end of the column body. 13. The pipette tip extraction column of claim 12 wherein the top frit has a pore volume of less than one microliter. 14. The pipette tip extraction column of claim 13 wherein the top frit or the bottom frit has a pore volume of less than 0.5 microliters. 15. The pipette tip extraction column of claim 12, wherein the top frit or the bottom frit comprises a membrane screen. 16. The pipette tip extraction column of claim 15, wherein the top frit or the bottom frit is comprised of a material selected from the group consisting of nylon, polyester, polyamide, polycarbonate, cellulose, polyethylene, nitrocellulose, cellulose acetate, polyvinylidene difluoride, polytetrafluoroethylene (PTFE), polypropylene, polysulfone, metal and glass. 17. The pipette tip extraction column of claim 15, wherein the top frit or the bottom frit comprises a nylon or polyester woven membrane. 18. A multiplexed extraction system comprised of: a plurality of pipette tip extraction columns of claim 1, wherein each extraction column is controlled by a pump. 19. The multiplexed extraction system of claim 18, wherein the multiplexed extraction system is further comprised of: a controller for controlling the pumps; anda computer operating said controller, wherein the computer can be programmed to control the movement of the pumps through the controller. 20. The multiplexed extraction system of claim 19, wherein the pipette tip extraction columns are further comprised of a top frit extending across the open channel between the bed of resin media and the open upper end of the column body, wherein the bottom frit is located at the open lower end of the column body, wherein the resin media is further comprised of an affinity binding group selected from the group consisting of Protein A, Protein G, Protein L and an immobilized metal. 21. The multiplexed extraction system of claim 19, wherein the bed of resin media is comprised of silica. 22. A pipette tip extraction column comprising: i) a column body having an open upper end, an open lower end, and an open channel between the upper and lower ends of the column body, wherein the column body is comprised of a modified pipette tip;ii) a bottom frit extending across the open channel, said bottom frit having a pore volume of less than one microliter, wherein the bottom frit is located at the open lower end of the column body;iii) a top frit extending across the open channel between the bottom frit and the open upper end of the column body; andiv) a bed of resin media positioned between the bottom frit and the top frit, wherein the resin media is further comprised of an affinity binding group selected from the group consisting of Protein A, Protein G, Protein L and an immobilized metal.
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