Hydrophobic interaction protein chromatography under no-salt conditions

Abstract

Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

1 .- 9 . (canceled) 
     
     
         10 . A method of purifying an antibody, the method comprising:
 subjecting an antibody in solution to hydrophobic interaction chromatography in flow through mode at a loading density of at least 70 g/L using a matrix containing hydrophobic ligands with a mobile phase buffer, wherein the mobile phase buffer does not contain a kosmotropic salt and has a pH of about 5.0 to about 7.0.   
     
     
         11 . The method of  claim 10 , wherein the loading density is between 70 g/L and 100 g/L. 
     
     
         12 . The method of  claim 10 , wherein the loading density is between 100 g/L and 200 g/L. 
     
     
         13 . The method of  claim 10 , wherein the loading density is between 150 g/L and 200 g/L. 
     
     
         14 . The method of  claim 10 , wherein the pH of the mobile phase buffer is about 5.0 to about 6.5, about 5.0 to about 6.0, about 5.5 to about 7.0, or about 6.0 to about 7.0. 
     
     
         15 . The method of  claim 10 , wherein the matrix comprises a hydroxylated methacrylic polymer, agarose or sepharose. 
     
     
         16 . The method of  claim 10 , wherein the hydrophobic ligands are ether groups, polypropylene glycol groups, phenyl groups, butyl groups, hexyl groups or octyl groups. 
     
     
         17 . The method of  claim 10 , wherein the antibody is a monoclonal antibody or a polyclonal antibody. 
     
     
         18 . The method of  claim 17 , wherein the antibody is a human antibody, a mouse antibody or a chimeric antibody. 
     
     
         19 . The method of  claim 10 , further comprising collecting a flow through fraction that contains the antibody. 
     
     
         20 . The method of  claim 10 , wherein the hydrophobic ligands comprise straight chain alkyl ligands or aryl ligands. 
     
     
         21 . The method of  claim 10 , wherein the matrix is a polymeric matrix. 
     
     
         22 . The method of  claim 10 , wherein the kosmotropic salt is selected from the group consisting of ammonium sulfate, sodium citrate, and potassium phosphate. 
     
     
         23 . A method of purifying an antibody, the method comprising:
 providing an antibody in a solution;   loading the solution at a loading density of at least 70 g/L onto a matrix containing hydrophobic ligands;   adding a mobile phase buffer that has a pH of about 5.0 to about 7.0, wherein the mobile phase buffer does not contain a kosmotropic salt; and   collecting a flow through fraction that contains the antibody.   
     
     
         24 . The method of  claim 23 , wherein the loading density is between 70 g/L and 100 g/L. 
     
     
         25 . The method of  claim 23 , wherein the loading density is between 100 g/L and 200 g/L. 
     
     
         26 . The method of  claim 23 , wherein the loading density is between 150 g/L and 200 g/L. 
     
     
         27 . The method of  claim 23 , wherein the kosmotropic salt is selected from the group consisting of ammonium sulfate, sodium citrate, and potassium phosphate. 
     
     
         28 . The method of  claim 23 , wherein the matrix comprises a hydroxylated methacrylic polymer, agarose or sepharose. 
     
     
         29 . The method of  claim 23 , wherein the hydrophobic ligands are ether groups, polypropylene glycol groups, phenyl groups, butyl groups, hexyl groups or octyl groups.