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.

What is claimed is: 
     
         1 . 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, wherein the core entity and the lid are made of agarose, and wherein the hydrophobic core entity is functionalized with hydrophobic interaction ligands which comprise C4-C16 alkyl aliphatic hydrocarbons. 
     
     
         2 . Separation medium according to  claim 1 , wherein the hydrophobic interaction ligands comprise C8 alkyl aliphatic hydrocarbons. 
     
     
         3 . Separation medium according to  claim 1 , wherein the ligand density is greater than 90 micromoles per millilitre of core entity. 
     
     
         4 . Separation medium according to  claim 1 , for use in purification of a target molecule whereby the target molecule is obtained in the flow-through, wherein the target molecule is:
 (i) monoclonal antibodies, or   (ii) virus.   
     
     
         5 . Separation medium according to  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. 
     
     
         6 . Separation medium according to  claim 1 , wherein the hydrophobic ligands are octyl ligands. 
     
     
         7 . Separation medium according to  claim 6 , wherein the hydrophobic ligands are octylamine. 
     
     
         8 . Separation medium according to  claim 1 , wherein the hydrophilic lid is a non-binding lid. 
     
     
         9 . Separation medium according to  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. 
     
     
         10 . Separation medium according to  claim 1 , wherein magnetic particles are incorporated into the core entities. 
     
     
         11 . Separation medium according to  claim 1 , wherein the lid and/or core entity are functionalized by layer activation. 
     
     
         12 . Separation medium according to  claim 1 , which is mixed with chromatography media, wherein the separation media comprises up to 10% of the total media volume. 
     
     
         13 . Separation medium according to  claim 12 , wherein the separation medium comprises octyl ligands and the chromatography media is a cation exchange media.