Process for the purification of antibodies

• US 20070167613A1
• Filed: 02/25/2005
• Published: 07/19/2007
• Est. Priority Date: 02/27/2004
• Status: Active Grant

• Current Assignee: Cytiva Bioprocess R&D AB (Danaher Corporation)
• Original Assignee: GE Healthcare Bio-Sciences AB (General Electric Company)
• Inventors: Johansson, Bo-Lennart, Maloisel, Jean-Luc, Johansson, Hans, Gronberg, Anna, Thevenin, Nicolas
• Granted Patent: US 7,750,129 B2
• Time in Patent Office: Days
• US Class Current: 530/389.100
• CPC Class Codes: B01D 15/3847 Multimodal interactions
• CPC Class Codes: B01J 20/3242 Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
• CPC Class Codes: B01J 20/3246 having a well defined chemical structure
• CPC Class Codes: B01J 20/3253 comprising a cyclic structure not containing any of the heteroatoms nitrogen, oxygen or sulfur, e.g. aromatic structures
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 20/3285 Coating or impregnation layers comprising different type of functional groups or interactions, e.g. different ligands in various parts of the sorbent, mixed mode, dual zone, bimodal, multimodal, ionic or hydrophobic, cationic or anionic, hydrophilic or hydrophobic

Abstract

The present invention relates to a process for the purification of antibodies from one or more impurities in a liquid, which process comprises contacting said liquid with a first chromatography resin comprised of a support to which multi-modal ligands have been immobilised to adsorb the antibodies to the resin, wherein each multi-modal ligand comprises at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system; adding an eluent to release the antibodies from the resin; and contacting the eluate so obtained with a second chromatography resin. In one embodiment, the ring-forming atoms of the aromatic or heteroaromatic entity are selected from the group consisting of C, S and O, and the cation exchanging group is a weak cation exchanger.

Claims

1. A method for capture of one or more antibodies from a liquid, comprising contacting said liquid with a chromatography resin including a support to which multi-modal ligands have been immobilised to adsorb the antibodies to the resin, wherein each multi-modal ligand includes at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system, in which system the ring-forming atoms are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.

2. The method of claim 1, wherein the ring-forming atoms are selected from the group consisting of carbon (C) and sulphur (S) atoms.

3. The method of claim 2, wherein the ring-forming atoms are carbon (C) atoms.

4. The method of claim 1, which is followed by one or further purification steps.

26. The method of claim 1, wherein the liquid contacted with the multi-modal chromatography resin is a cell culture liquid or a fermentation broth.

27. The method of claim 1, wherein the cation-exchanging group of the multi-modal ligands is a weak cation exchanger.

28. The method of claim 1, wherein the antibodies are monoclonal antibodies.

29. The method of claim 1, wherein the antibodies are polyclonal antibodies.

5. A process for the purification of one or more antibodies from a liquid, comprising contacting said liquid with a first chromatography resin including a support to which multi-modal ligands have been immobilised to adsorb the antibodies to the resin, wherein each multi-modal ligand includes at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system;

adding an eluent to release the antibodies from the resin; and

contacting the eluate so obtained with a second chromatography resin.

6. The process of claim 5, wherein the liquid contacted with the multi-modal chromatography resin is a cell culture liquid or a fermentation broth.

7. The process of claim 5, wherein the ring-forming atoms of the aromatic or heteroaromatic ring system of the multi-modal ligands are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.

8. The process of claim 5, wherein the cation-exchanging group of the multi-modal ligands is a weak cation exchanger.

9. The process of claim 5, wherein the second chromatography step is selected from the group consisting of ion exchange chromatography;

hydrophobic interaction chromatography (HIC);

immobilised metal affinity chromatography (IMAC); and

affinity chromatography.

10. The process of claim 9, wherein the second chromatography step is ion exchange chromatography.

11. The process of claim 5, wherein the second chromatography step is anion exchange chromatography.

12. The process of claim 5, wherein the second chromatography step is multimodal anion exchange chromatography.

13. The process of claim 5, wherein antibodies are recovered from the flow-through of the second chromatography resin.

14. The process of claim 5, wherein antibodies and/or impurities are eluted from the second chromatography resin.

15. The process of claim 5, wherein the antibodies are monoclonal antibodies.

16. The process of claim 5, wherein the antibodies are polyclonal antibodies.

17. A method for capture of one or more antibodies from a liquid, comprising contacting said liquid with a chromatography resin to adsorb the antibodies to the ligands, which resin is multimodal and comprises a support to which ligands have been immobilised, wherein the resin includes cation-exchanging groups and aromatic or heteroaromatic ring systems present on the same or different ligands, in which systems the ring-forming atoms are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.

18. A process for the purification of one or more antibodies from a liquid, comprising contacting said liquid with a first chromatography resin to adsorb the antibodies to the ligands, which resin is multimodal and comprises a support to which ligands have been immobilised, wherein the resin includes cation-exchanging groups and aromatic or heteroaromatic ring systems present on the same or different ligands;

adding an eluent to release the antibodies from the resin; and

contacting the eluate so obtained with a second chromatography resin.

19. A kit for the purification of one or more antibodies in a liquid, comprising in separate compartments a first chromatography column packed with a chromatography resin, which resin is multimodal and includes a support to which ligands have been immobilised, wherein cation-exchanging groups and aromatic or heteroaromatic ring systems are present on the same or different ligands;

a second chromatography column packed with a chromatography resin;

one or more buffers for adsorption and or elution of antibodies; and

written instructions that teaches the purification of antibodies in a two-step process.

20. The kit of claim 19, wherein the ring-forming atoms of the aromatic or heteroaromatic ring system of the multi-modal ligands are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.

21. The kit of claim 19, wherein the first chromatography column is a sterile column.

22. The kit of claim 19, wherein the first chromatography column is a disposable column.

23. A disposable chromatography column for the purification of antibodies, comprising a multi-modal chromatography resin comprising cation-exchanging groups and aromatic and/or heteroaromatic ring systems on the same or different ligands.

25. The disposable column of claim 23, wherein the ring-forming atoms of the aromatic or heteroaromatic ring system of the multi-modal ligands are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.

24. A disposable chromatography column for the purification of antibodies, comprising a multi-modal chromatography resin, wherein each ligand comprises at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system.

30. The disposable column of claim 24, wherein the ring-forming atoms of the aromatic or heteroaromatic ring system of the multi-modal ligands are selected from the group consisting of carbon (C), sulphur (S) and oxygen (O) atoms.