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 between two frits. In some embodiments, the extraction columns employ modified pipette tips as column bodies. In some embodiments, the invention provides methods of storing columns in a wet state.
대표청구항▼
1. A method of producing a pipette tip column, comprised of: a) providing a pipette tip, having an open upper end and an open lower end;b) providing a screen frit;c) attaching the screen frit to the open lower end of the pipette tip to yield a lower frit;d) providing a gel resin;e) introducing the g
1. A method of producing a pipette tip column, comprised of: a) providing a pipette tip, having an open upper end and an open lower end;b) providing a screen frit;c) attaching the screen frit to the open lower end of the pipette tip to yield a lower frit;d) providing a gel resin;e) introducing the gel resin into the open upper end of the pipette tip to produce a bed of medium positioned above the lower frit, wherein the bed of medium is comprised of an interstitial space;f) providing a water-miscible solvent, wherein the solvent has a boiling point of at least 150 degrees C, wherein the solvent content of the water-miscible solvent is at least 75%;g) introducing the water-miscible solvent into the bed of medium subsequent to introduction of the gel resin, whereby the water-miscible solvent occupies the entire interstitial space; andh) storing the column at a temperature between 20 and 30 degrees C for a storage period, wherein the storage period is at least 24 hours, wherein the solvent remains in the bed of medium during the storage period. 2. The method of claim 1, wherein multiple pipette tip columns are produced and stored. 3. The method of claim 2, wherein the pipette tip columns are packaged in a rack. 4. The method of claim 3, wherein the rack holds 96 pipette tip columns. 5. The method of claim 1, wherein the water-miscible solvent is comprised of glycerol or ethylene glycol. 6. The method of claim 1, wherein the water-miscible solvent has a boiling point of at least 190 degrees centigrade. 7. The method of claim 1, wherein the column is not capped during the storage period. 8. The method of claim 1, wherein the solvent content of the water-miscible solvent is at least 85%. 9. The method of claim 8, wherein the solvent content of the water-miscible solvent is at least 95%. 10. The method of claim 1, wherein the column is stored at ambient air pressure and humidity. 11. The method of claim 1, wherein following the storage period, the column is shipped to an end user. 12. The method of claim 1, wherein following the storage period, at least 90% of the interstitial space is occupied by the solvent. 13. The method of claim 12, wherein following the storage period, at least 95% of the interstitial space is occupied by the solvent. 14. The method of claim 13, wherein following the storage period, at least 98% of the interstitial space is occupied by the solvent. 15. The method of claim 1, wherein the gel resin is hydrated. 16. The method of claim 15, wherein the hydrated gel resin is further comprised of an affinity binding agent. 17. The method of claim 16, wherein the affinity binding agent is an antibody. 18. The method of claim 1, wherein the lower frit is bonded to the column body. 19. The method of claim 1, wherein between steps (e) and (f), a top frit is attached to the pipette tip column above the bed of medium. 20. The method of claim 19, wherein after the top frit is attached, the pipette tip column has a back pressure in the range of 0.01 to 2 pounds per square inch at an aqueous flow rate of 1 ml/min. 21. A method of producing a pipette tip column, comprised of: a) providing a pipette tip, having an open upper end and an open lower end;b) providing a screen frit;c) attaching the screen frit to the open lower end of the pipette tip to yield a lower frit;d) providing a gel resin;e) introducing the gel resin into the open upper end of the pipette tip to produce a bed of medium positioned above the lower frit, wherein the bed of medium is comprised of an interstitial space;f) providing a water-miscible solvent, wherein the solvent has a boiling point of at least 150 degrees C, wherein the solvent content of the water-miscible solvent is at least 75%;g) introducing the water-miscible solvent into the bed of medium subsequent to introduction of the gel resin, whereby the water-miscible solvent occupies the entire interstitial space; andh) shipping the column to an end user, wherein the column is shipped at ambient temperature and wherein the solvent remains in the bed of medium. 22. The method of claim 21, wherein multiple pipette tip columns are produced and shipped. 23. The method of claim 22, wherein the pipette tip columns are packaged in a rack. 24. The method of claim 23, wherein the rack holds 96 pipette tip columns. 25. The method of claim 21, wherein the water-miscible solvent is comprised of glycerol or ethylene glycol. 26. The method of claim 21, wherein the water-miscible solvent has a boiling point of at least 190 degrees centigrade. 27. The method of claim 21, wherein the column is not capped during shipping. 28. The method of claim 21, wherein the solvent content of the water-miscible solvent is at least 85%. 29. The method of claim 28, wherein the solvent content of the water-miscible solvent is at least 95%. 30. The method of claim 21, wherein the column is shipped at ambient air pressure and humidity. 31. The method of claim 21, wherein after the column is shipped, at least 90% of the interstitial space is occupied by the solvent. 32. The method of claim 31, wherein after the column is shipped, at least 95% of the interstitial space is occupied by the solvent. 33. The method of claim 32, wherein after the column is shipped, at least 98% of the interstitial space is occupied by the solvent. 34. The method of claim 21, wherein the gel resin is hydrated. 35. The method of claim 21, wherein the hydrated gel resin is further comprised of an affinity binding agent. 36. The method of claim 35, wherein the affinity binding agent is an antibody. 37. The method of claim 21, wherein the lower frit is bonded to the column body. 38. The method of claim 21, wherein between steps (e) and (f), a top frit is attached to the pipette tip column above the bed of medium. 39. The method of claim 38, wherein after the top frit is attached, the pipette tip column has a back pressure in the range of 0.01 to 2 pounds per square inch at an aqueous flow rate of 1 ml/min. 40. A method of producing a pipette tip column, comprised of: a) providing a pipette tip, having an open upper end and an open lower end;b) providing a screen frit;c) attaching the screen frit to the open lower end of the pipette tip to yield a lower frit;d) providing resin beads, wherein the resin beads are comprised of an affinity binding group and wherein the affinity binding group is an antibody;e) introducing the resin beads into the open upper end of the pipette tip to produce a bed of medium positioned above the lower flit, wherein the bed of medium is comprised of an interstitial space;f) providing a water-miscible solvent, wherein the solvent has a boiling point of at least 150 degrees C, wherein the solvent content of the water-miscible solvent is at least 75%;g) introducing the water-miscible solvent into the bed of medium subsequent to introduction of the resin beads, whereby the water-miscible solvent occupies the entire interstitial space; andh) storing the column at a temperature between 20 and 30 degrees C for a storage period, wherein the storage period is at least 24 hours, wherein the solvent remains in the bed of medium during the storage period. 41. The method of claim 40, wherein multiple pipette tip columns are produced and stored. 42. The method of claim 41, wherein the pipette tip columns are packaged in a rack. 43. The method of claim 42, wherein the rack holds 96 pipette tip columns. 44. The method of claim 40, wherein the water-miscible solvent is comprised of glycerol or ethylene glycol. 45. The method of claim 40, wherein the water-miscible solvent has a boiling point of at least 190 degrees centigrade. 46. The method of claim 40, wherein the column is not capped during the storage period. 47. The method of claim 40, wherein the solvent content of the water-miscible solvent is at least 85%. 48. The method of claim 47, wherein the solvent content of the water-miscible solvent is at least 95%. 49. The method of claim 40, wherein the column is stored at ambient air pressure and humidity. 50. The method of claim 40, wherein following the storage period, the column is shipped to an end user. 51. The method of claim 40, wherein following the storage period, at least 90% of the interstitial space is occupied by the solvent. 52. The method of claim 51, wherein following the storage period, at least 95% of the interstitial space is occupied by the solvent. 53. The method of claim 52, wherein after the column is shipped, at least 98% of the interstitial space is occupied b the solvent. 54. The method of claim 40, wherein the lower frit is bonded to the column body. 55. The method of claim 40, wherein between steps (e) and (f), a top frit is attached to the pipette tip column above the bed of medium. 56. The method of claim 55, wherein after the top frit is attached, the pipette tip column has a back pressure in the range of 0.01 to 2 pounds per square inch at an aqueous flow rate of 1 ml/min. 57. The method of claim 40, wherein the resin beads are comprised of a pellicular resin.
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이 특허에 인용된 특허 (23)
Wainwright Norman B. (Falmouth MA) Boyd Steven H. (Falmouth MA), Activated immunodiagnostic pipette tips.
Raybuck Margaret P. (Pontyclun Mid Glamorgan GBX) Kenrick Michael K. (Cardiff GBX) Parry David A. (London GBX), Device and method for affinity separation.
Magnussen ; Jr. Haakon T. (Orinda CA) Ruskewicz Stephen J. (Kensington CA) Smith Gary L. (Walnut Creek CA) Wingo Anthony K. (San Leandro CA), Method for dispensing liquids with a pipette with compensation for air pressure and surface tension.
Bodo Gerhard (Vienna) Maurer-Fogy Ingrid (Vienna) Falkner Edgar (Vienna) Lindner Silvia J. (Perchtoldsdorf ATX), Process for preparing and purifying alpha-interferon.
Lemieux David J. (Middleton MA) Chadwick Paul K. (San Francisco CA) Kelly Christopher (Larkspur CA) Petrek James S. (Danville CA) Rainin Kenneth (Piedmont CA) Magnussen ; Jr. Haakon T. (Orinda CA) Ho, Refill pack for pipette tip racks.
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