Chromatography ligand
View Patent ↗The present invention relates to a chromatography ligand defined by the following formula R 1 —R 2 —N(R 3 )—R 4 —R 5 wherein R 1 is a substituted or non-substituted phenyl group; R 2 is a hydrocarbon chain comprising 0-4 carbon atoms; R 3 is a hydrocarbon chain comprising 1-3 carbon atoms; R 4 is a hydrocarbon chain comprising 1-5 carbon atoms; and R 5 is OH or H. The invention also comprises a separation matrix, comprising the described ligands coupled to a porous support, such as particles or a membrane. The ligand and matrix according to the invention is useful for purification of biomolecules or organic compounds, such as proteins, polypeptides, DNA etc. An advantageous use according to the invention is the purification of antibodies.
1. A method of separating one or more antibodies from one or more other compounds in a liquid sample, wherein a mobile phase comprising said antibodies and compound(s) is contacted with a separation matrix, wherein said separation matrix comprises a chromatography ligand defined by the following formula:
R 1 —R 2 —N(R 3 )—R 4 —R 5
wherein
R 1 is a non-substituted phenyl group;
R 2 is —CH 2 —;
R 3 is —CH 3 ;
R 4 is a hydrocarbon chain comprising 1-5 carbon atoms; and
R 5 is OH, and wherein said chromatography ligand is immobilized to a support via the amine nitrogen atom, wherein said compounds comprise host cell proteins, protein A, or antibody aggregates and wherein the host cell proteins, protein A, or antibody aggregates are separated from the antibodies.
2. The method of claim 1 , wherein the separation matrix is provided in a chromatography column, the mobile phase is passed across 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 separation matrix is preceded by a step of mechanical filtration and/or chromatography.
5. The method of claim 1 , wherein the liquid sample comprises a crude feed.
6. The method of claim 1 , wherein the adsorbed compound(s) are host cell proteins and substantially all said proteins are adsorbed to the separation matrix.
7. The method of claim 1 , wherein the liquid sample is an eluate from a preceding affinity chromatography matrix.
8. The method of claim 1 , wherein the antibodies have a pH in the range of from 6 to about 9.
9. The method of claim 1 , wherein at least 95% of the contaminants are removed on the matrix.
10. The method of claim 1 , wherein the ligand is selected from the group consisting of N-benzyl-N-methyl ethanolamine.
11. The method of claim 1 , wherein said compounds are adsorbed to the separation matrix.