Purification of biological products by constrained cohydration chromatography

Abstract

Materials and methods for use of constrained cohydration agents in the purification of biological materials such as antibodies, viruses, cells, and cellular organelles in connection with convective chromatography, fluidized bed or co-precipitation applications.

1 . A method for purification of a target species of biological origin from a sample comprising:
 (a) providing a monolith comprising a convective chromatography material comprising a hydrated surface, the hydrated surface comprising charged moieties;   (b) contacting the sample comprising the target species with the hydrated surface in the presence of,
 (i) an amount of a non-ionic organic polymer sufficient to cause more than 50% to substantially all of the target species to be retained at the hydrated surface exclusively by constrained cohydration, and 
 (ii) a buffer providing conditions of pH, conductivity or a salt concentration that provide a substantial absence of a direct chemical interaction between the target species and the hydrated surface. 
   
     
     
         2 . The method of  claim 1 , wherein the non-ionic organic polymer comprises polyethylene glycol. 
     
     
         3 . The method of  claim 2 , wherein the polyethylene glycol has an average polymer weight between 100 and 10,000 Daltons. 
     
     
         4 . The method of  claim 3 , wherein the polyethylene glycol has an average polymer weight between 4000 and 8,000 Daltons. 
     
     
         5 . The method of  claim 1 , wherein the amount of the non-ionic organic polymer is between approximately 2% and 50% (w/v). 
     
     
         6 . The method of  claim 5 , wherein the amount of the non-ionic organic polymer is between approximately 15% and 25% (w/v). 
     
     
         7 . The method of  claim 1 , wherein the monolith comprises polyacrylate, polymethacrylate, styrenedivinylbenzene or silica. 
     
     
         8 . The method of  claim 1 , wherein the monolith has an average channel size between about 1 micron and 200 microns. 
     
     
         9 . The method of  claim 8 , wherein the average channel size is between about 1 micron and 20 microns. 
     
     
         10 . The method of  claim 1 , wherein the chemical moieties comprise a cationic charged group or an anionic charged group. 
     
     
         11 . The method of  claim 1 , wherein the chemical moieties comprise a polar moiety selected from the group consisting of hydroxyl, amine, imine, ureide, carbohydrate, and amino acid. 
     
     
         12 . The method of  claim 11 , wherein the chemical moieties comprise a hydroxyl group. 
     
     
         13 . The method of  claim 1 , wherein the target species comprises a virus, an antibody, a protein, or exosomes. 
     
     
         14 . The method of  claim 13 , wherein the target species comprises a virus. 
     
     
         15 . The method of  claim 13 , wherein the sample is a virus-containing solution derived from a cell culture. 
     
     
         16 . The method of  claim 13 , wherein the target species comprises a monoclonal antibody. 
     
     
         17 . The method of  claim 16 , wherein the target species comprises an IgM monoclonal antibody. 
     
     
         18 . The method of  claim 1 , further comprising:
 (c) dissociating the target species from the hydrated surface by contacting the hydrated surface of the monolith with a dissociation buffer comprising an amount of the non-ionic organic polymer that is less than 2% percent (w/v).   
     
     
         19 . The method of  claim 18  comprising:
 after the contacting of (b), by prior to the dissociating of (c), washing the monolith with a solution comprising the non-ionic organic polymer wherein more than 50% to substantially all of the target species remains retained at the hydrated surface. 
 
     
     
         20 . The method of  claim 1 , wherein prior to (b), the monolith is equilibrated with a solution comprising the constraining agent.