Chromatography medium
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. An agarose-based separation medium, comprising,
a hydrophobic porous core entity with octylamine ligands having a ligand density of greater than 90 μmole/mL; and
a porous hydrophilic lid covering the whole outer part of the core entity,
wherein the lid only allows molecules under 60,000D to penetrate and absorb onto the hydrophobic octyl ligands of said core entity of the separation medium,
wherein the hydrophobic core entity allows adsorption of molecules under 60,000D at low ionic strength 0-1M in the pH interval 2-11.
2. The separation medium of claim 1 , wherein the hydrophobic core entity allows adsorption of molecules under 1,000D.
3. The separation medium of claim 1 , wherein the hydrophilic lid comprises ligands that reversibly bind the molecules within a sample.
4. The separation medium of claim 1 , wherein magnetic particles are incorporated into the core entity.
5. The separation medium of claim 1 , wherein a dextran hydrogel is provided in the lid to fill the pores of the lid and thereby further decrease and adjust the pore size of the lid.
6. The separation medium of claim 1 , wherein at least one of the lid and core entity are functionalized by layer activation.