Method of purifying an antibody
A method of purifying an antibody composition comprises application of anion exchange chromatography late in the purification process. An ultrafiltration/diafiltration-purified antibody composition is subjected to anion exchange chromatography (AEX) to form a pharmaceutically-pure antibody composition.
1. A method of purifying an antibody composition, wherein the antibody is produced in eukaryotic cells, which comprises subjecting a UF/DF-purified antibody composition to anion exchange chromatography (AEX) to form a pharmaceutically-pure antibody composition, wherein said anion exchange chromatography is the last chromatographic purification step and is performed directly after the final UF/DF step.
2. The method of claim 1 , wherein said UF/DF-purified antibody composition has an antibody concentration of at least 1 mg/ml.
3. The method of claim 2 , wherein said UF/DF-purified antibody composition has an antibody concentration of at least 10 mg/ml.
4. The method of claim 3 , wherein said UF/DF-purified antibody composition has an antibody concentration of not greater than 250 mg/ml.
5. The method of claim 4 , wherein said UF/DF-purified antibody composition has an antibody concentration of 20 to 50 mg/ml.
6. The method of claim 1 , which further comprises filling said pharmaceutically-pure antibody composition into vials.
7. The method according to claim 6 , which further comprises aseptically filtering said pharmaceutically-pure antibody composition prior to said filling step.
8. The method according to claim 7 , which further comprises at least one of the following steps prior to said filling step: (a) adding an excipient to said pharmaceutically-pure antibody composition; (b) concentrating said pharmaceutically-pure antibody composition; (c) diluting said pharmaceutically-pure antibody composition; and/or (d) adjusting the pH of said pharmaceutically-pure antibody composition.
9. The method of claim 1 , wherein said UF/DF-purified antibody composition is obtained by subjecting a partially-purified antibody composition to UF/DF.
10. The method of claim 9 , wherein an antibody cell culture harvest is subjected to an antibody capture step and optionally at least one polishing step to form said partially-purified antibody composition.
11. The method of claim 10 , wherein said capture step utilizes affinity chromatography.
12. The method of claim 11 , wherein said affinity chromatography is Protein A chromatography.
13. The method of claim 10 , wherein said capture step utilizes cation exchange chromatography (CEX), hydrophobic charge induction chromatography (HCIC), or mixed mode chromatography.
14. The method of claim 10 , wherein at least one polishing step is carried out after said capture step.
15. The method according to claim 14 , wherein said at least one polishing step is selected from the group consisting of ion exchange chromatography (IEX), hydrophobic interaction chromatography (HIC), and hydroxyapatite chromatography.
16. The method of claim 1 , wherein said antibody is trastuzumab or pertuzumab.
17. The method of claim 1 , wherein said antibody was produced in CHO cells.
18. A method of purifying an antibody composition, which comprises:
subjecting an antibody eukaryotic cell culture harvest to affinity chromatography to capture a crude antibody composition;
subjecting the crude antibody composition to at least one polishing step to form a partially-purified antibody composition;
subjecting the partially-purified antibody composition to a UF/DF step to form a UF/DF-purified antibody composition; and
subjecting the UF/DF-purified antibody composition to anion exchange chromatography (AEX) to form a pharmaceutically-pure antibody composition,
wherein said anion exchange chromatography is the last chromatographic purification step and is performed directly after the final UF/DF step.
19. The method of claim 18 , wherein said affinity chromatography is Protein A chromatography.
20. The method of claim 18 , which further comprises subjecting said pharmaceutically-pure antibody composition to a virus removal step.
21. The method of claim 20 , wherein said virus removal step is nanofiltration.
22. The method of claim 18 , wherein said at least one polishing step comprises subjecting said crude antibody composition to at least one ionic exchange chromatography (IEX) step.