Method of sterilization
View Patent ↗The present invention relates to a method of sterilizing a chromatography column which is packed with a separation matrix, which method comprises passing a superheated aqueous liquid through the packing of the column to obtain at least one substantially sterile packed chromatography column. The method may also comprise a preceding heating. In the most advantageous embodiment, the superheated aqueous liquid is passed in a substantially uniform flow through the packing of the column, resulting in a sterility of the packed column.
1. A method of sterilizing at least one chromatography column packed with a separation matrix, comprising:
providing a sealed chromatography column made from a material which withstands the temperature and pressure conditions used;
applying a pressure over the chromatography column;
heating an aqueous liquid which is a buffer commonly used in chromatography to a temperature which under the prevailing pressure provides a superheated aqueous liquid; and
passing said superheated aqueous liquid through the packing of said column(s) to obtain at least one substantially sterile packed chromatography column, wherein the temperature and pressure of the superheated aqueous liquid are substantially maintained throughout the column such that said aqueous liquid is maintained superheated.
2. The method of claim 1 , wherein the superheated aqueous liquid is passed through the packing of the column in a substantially uniform flow.
3. The method of claim 1 , wherein the superheated aqueous liquid is added to an inlet at one end of the packed chromatography column and withdrawn at an outlet located substantially opposite to the inlet.
4. The method of claim 1 , wherein the superheated aqueous liquid is added to the centre of the packed chromatography column and withdrawn at the periphery of the packed chromatography column.
5. The method of claim 1 , wherein the liquid is pumped from a heater to the chromatography column and/or through the chromatography column.
6. The method of claim 1 , further comprising monitoring the aqueous liquid temperature at an outlet of the column.
7. The method of claim 6 , further comprising controlling the temperature at an inlet into the column based on the temperature at the outlet.
8. The method of claim 1 , wherein the matrix is comprised of porous particles.
9. The method of claim 1 , wherein the matrix is comprised of cross-linked polysaccharide particles.
10. The method of claim 1 , wherein the matrix is comprised of synthetic polymer particles.
11. The method of claim 1 , wherein the matrix is comprised of monolithic material.
12. The method of claim 8 , wherein the surfaces of said porous particles present at least one ligand.
13. The method of claim 12 , wherein the ligand(s) are substantially devoid of heat-sensitive peptide bonds.
14. The method of claim 12 , wherein the ligand(s) are selected from the group consisting of ion-exchange ligands, multimodal ligands, thiophilic ligands and hydrophobic chromatography (HIC) ligands.