Method of antibody purification
View Patent ↗The present invention relates to a method of separating antibodies from other compound(s) in a liquid sample, wherein a mobile phase comprising said sample is contacted with a multi-modal separation matrix to adsorb undesired compounds while the antibodies remain free in the liquid, wherein the multi-modal separation matrix comprises first groups, which are capable of interacting with negatively charged sites of the target compounds, and second groups, which are capable of at least one interaction other than charge-charge interaction with said target compounds. The invention also relates to a chromatography column packed with the above-described multi-modal separation matrix and a filter having such multi-modal groups adsorbed to its surface.
1. A method for separating one or more antibodies from one or more other compounds in a liquid sample, which method comprises contacting a mobile phase comprising said liquid sample with a multi-modal separation matrix to adsorb one or more of said other compounds while the antibodies remain free in the mobile phase, wherein the multi-modal separation matrix comprises first groups capable of interacting with negatively charged sites of said other compound(s) and second groups capable of at least one interaction other than charge-charge interaction with said other compound(s).
2. The method of claim 1 , wherein the multi-modal separation matrix is provided in a chromatography column, the mobile phase is passed through said column by gravity and/or pumping, and the antibodies are recovered in the flow-through of the column.
3. The method of claim 1 , wherein the liquid sample comprises a supernatant obtained from cell fermentation.
4. The method of claim 1 , wherein the contact with the multi-modal separation matrix is preceded by a step of mechanical filtration and/or chromatography.
5. The method of claim 4 , wherein the liquid sample comprises an eluate from a separation matrix.
6. The method of claim 5 , wherein the separation matrix from which the eluate is obtained comprises protein ligands.
7. The method of claim 5 , wherein the separation matrix from which the eluate is obtained comprises protein A or G ligands.
8. The method of claim 1 , wherein the liquid sample comprises a crude feed.
9. The method of claim 8 , wherein the other compound(s) are host cell proteins and substantially all said proteins are adsorbed to the multi-modal separation matrix.
10. The method of claim 1 , wherein the conductivity of the mobile phase is in the range of 0-25 mS/cm.
11. The method of claim 1 , wherein the first groups are quaternary amines.
12. The method of claim 1 , wherein the second groups are hydrogen-bonding groups.
13. The method of claim 1 , wherein the second groups are hydrophobic groups.
14. The method of claim 1 , wherein the separation matrix comprises first and second groups coupled to same ligands which are immobilized on said matrix.
15. The method of claim 14 , wherein the first and second group are distanced from each other by a hydrocarbon chain of 1-3 carbon atoms.
16. The method of claim 1 , wherein ligands have been immobilised to a support of said separation matrix via said first groups.
17. The method of claim 1 , wherein the separation matrix comprises first and second groups coupled to different ligands which are immobilized on said matrix.
18. The method of claim 1 , wherein the separation matrix is particulate and comprises a mixture of first particles, to which ligands comprising the first groups have been immobilised; and second particles, to which ligands comprising the second groups have been immobilised.
19. The method of claim 1 , wherein the separation matrix is a filter to which a mixture of first ligands comprising the first groups; and second ligands comprising the second groups have been immobilised.
20. The method of claim 1 , wherein the separation matrix comprises third groups capable of a third interaction with one said other compound.
21. The method of claim 1 , wherein the antibodies are monoclonal antibodies.
22. The method of claim 21 , wherein the antibodies are humanised antibodies.
23. The method of claim 1 , wherein the multi-modal separation matrix is provided in a disposable chromatography column.
24. The method of claim 23 , wherein the disposable column is sterilised prior to contacting the mobile phase.
25. The method of claim 1 , wherein the second groups comprise aromatic or heteroaromatic ring structure(s).