Chromatography medium

Abstract

The present invention is within the field of chromatography. More precisely, it relates to a novel chromatography medium, namely a hydrophobic medium provided with different lids excluding molecules over a certain size due to the porosity of the hydrophobic medium and/or the porosity of the lid. The invention also relates to use of the separation medium for purification of large molecules, which do not enter the separation medium, as well as small molecules, which enter the separation medium and are eluted from there.

1 . A separation medium comprising a porous, hydrophobic core entity; and a porous hydrophilic lid covering the whole outer part of the core entity, wherein the lid only allows molecules under a certain size to penetrate and interact with the internal part of the core entity of the separation medium. 
     
     
         2 . The separation medium of  claim 1 , wherein the hydrophobic core entity only allows molecules under a certain size to penetrate and interact with the internal part of the core entity of the separation medium. 
     
     
         3 . The separation medium of  claim 1 , wherein the hydrophobic core entity is hydrophobic per se and is based on a hydrophobic polymer. 
     
     
         4 . The separation medium of  claim 1 , wherein the hydrophobic core entity is hydrophilic per se and based on a hydrophilic polymer, functionalized with hydrophobic interaction ligands. 
     
     
         5 . The separation medium of  claim 4 , wherein the hydrophobic interaction ligands comprise aliphatic or aromatic hydrocarbons. 
     
     
         6 . The separation medium of  claim 1 , wherein the hydrophobic core entity allows adsorption of low molecular weight molecules and/or small size molecules at low ionic strength 0-1M in the pH interval 2-11. 
     
     
         7 . The separation medium of  claim 4 , wherein the core entity comprises further ligands besides said hydrophobic ligands which may be located on and/or associated with the core entity and/or on the hydrophobic ligands. 
     
     
         8 . The separation medium of  claim 7 , wherein said further ligands are electrostatic interaction ligands. 
     
     
         9 . The separation medium of  claim 1 , wherein the core entity has a pore size preventing molecules over a certain size from entering the pores. 
     
     
         10 . The separation medium of  claim 1 , wherein the hydrophilic lid comprises ligands that reversibly bind the molecules within the sample. 
     
     
         11 . The separation medium of  claim 4 , wherein the core entity and lid are made of agarose and the hydrophobic ligands are C 4 -C 16  ligands. 
     
     
         12 . The separation medium of  claim 1 , wherein a hydrogel is provided in the lid to fill the pores and thereby further decrease and adjust the pore size to prevent high molecular weight molecules from entering the pores. 
     
     
         13 . The separation medium of  claim 1 , wherein magnetic particles are incorporated into the core entities. 
     
     
         14 . The separation medium of  claim 1 , wherein the lid and/or core entity are functionalized by layer activation. 
     
     
         15 . The separation medium of  claim 1 , which is mixed with chromatography media, wherein the separation media comprises up to 10% of the total media volume. 
     
     
         16 . The separation medium of  claim 15 , wherein the separation medium comprises octyl ligands and the chromatography media is a cation exchange media. 
     
     
         17 . A method for separation comprising the step of adsorbing molecules under a certain size from a sample, onto the separation medium of  claim 1 , at low ionic strength above or equal to 0M, at pH 2-11. 
     
     
         18 . The method for separation of  claim 17 , wherein the certain size corresponds to the size range represented by molecules/organisms/particles selected from cells, cell particles, bacteria, virus, virus like particles, plasmids, antibodies, lipids, proteins, peptides or nucleic acid. 
     
     
         19 . The method for separation of  claim 17 , wherein the target molecule is a small molecule or particle <60 000D. 
     
     
         20 . The method for separation of  claim 17 , wherein the target molecule is a molecule or particle <1 000 kD. 
     
     
         21 . The method for separation of  claim 17 , wherein the target molecule is a large molecule/organism/particle, such as a cell, cell particles, bacteria, virus, virus like particles, plasmids, antibodies, proteins, which is not adsorbed on the separation medium but obtained in the flow-through. 
     
     
         22 . The method for separation of  claim 17 , wherein the target molecule is a large molecule/organism/particle, such as a cell, cell particles, bacteria, virus, virus like particles, plasmids, antibodies, proteins, which is adsorbed on the lid.