Method and Device for Extracting an Analyte

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 maintaining the columns in a wet state. The columns typically include a bed of extraction media positioned in the column between two frits. In some embodiments, the extraction column body is comprised of a modified pipette tip.

1 . A method of maintaining a column bed in a wet state, comprising the steps of:
 (a) providing a column, wherein the column is comprised of a bed of hydrated gel resin, the interstital space of which is substantially full of a liquid, wherein said liquid is a water miscible solvent that has a boiling point of at least 150° C., whereby said liquid is provided in an amount sufficient to maintain the hydrated gel resin in a wet state; and   (b) storing the column so that it is open to ambient temperature, air pressure and humidity.   
     
     
         2 . The method of  claim 1 , wherein the water miscible solvent has a boiling point of at least 175° C. 
     
     
         3 . The method of  claim 1 , wherein said water miscible solvent comprises glycerol. 
     
     
         4 . The method of  claim 1 , wherein the gel resin is comprised of agarose or sepharose. 
     
     
         5 . The method of  claim 1 , wherein the gel resin beads are comprised of an affinity binding group. 
     
     
         6 . The method of  claim 5 , wherein the affinity binding group has an affinity for an active protein. 
     
     
         7 . The method of  claim 5 , wherein the affinity binding group is selected from the group consisting of Protein A, Protein G, Protein L and an immobilized metal. 
     
     
         8 . The method of  claim 1 , wherein the water miscible solvent is resistant to bacterial growth. 
     
     
         9 . The method of  claim 1 , wherein the column is stored for at least 5 minutes. 
     
     
         10 . The method of  claim 9 , wherein the column is stored for at least 24 hours. 
     
     
         11 . The method of  claim 10 , wherein the column is stored for at least 5 days. 
     
     
         12 . The method of  claim 11 , wherein the column is stored for at least 90 days. 
     
     
         13 . The method of  claim 12 , wherein the column is stored for at least 1 year. 
     
     
         14 . A column bed comprised of:
 a bed of hydrated gel resin the interstital space of which is substantially full of a liquid, wherein said liquid is a water miscible solvent that has a boiling point of at least 150° C., whereby said liquid is provided in an amount that is sufficient to maintain the hydrated gel resin in a wet state, and wherein the column bed is open to ambient temperature, air pressure and humidity.   
     
     
         15 . The column bed of  claim 14 , wherein the water miscible solvent has a boiling point of at least 175° C. 
     
     
         16 . The column bed of  claim 14 , wherein the water miscible solvent comprises glycerol or ethylene glycol. 
     
     
         17 . The column bed of  claim 14 , wherein the gel resin is comprised of agarose or sepharose. 
     
     
         18 . The column bed of  claim 14 , wherein the gel resin is comprised of an affinity binding group having an affinity for an active protein. 
     
     
         19 . The column bed of  claim 18 , wherein the affinity binding group is selected from the group consisting of Protein A, Protein G, Protein L and an immobilized metal. 
     
     
         20 . The column bed of  claim 14 , wherein the bed is positioned within a pipette tip column.