Isolation and purification of antibodies using protein a affinity chromatography

• US 9,018,361 B2
• Filed: 03/26/2014
• Issued: 04/28/2015
• Est. Priority Date: 10/20/2008
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbvie Incorporated
• Inventors: Hickman, Robert K., Huang, Qing, Weed, Cheryl L., Perilli-Palmer, Barbara, Wan, Min, Ennis, Scott T.
• Primary Examiner(s): YAKOVLEVA, GALINA M
• Application Number:
  US14/226,636
• Publication Number:
  US 20140255423A1
• Time in Patent Office: 398 Days
• Field of Search: None
• US Class Current: 530/413
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 1/165 mixed-mode chromatography
• CPC Class Codes: C07K 1/18 Ion-exchange chromatography
• CPC Class Codes: C07K 1/20 Partition-, reverse-phase or hydrophobic interaction chromatography
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C07K 1/36 by a combination of two or more processes of different types
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/22 from camelids, e.g. camel, llama or dromedary
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

Disclosed herein are methods for the isolation and purification of antibodies wherein the use of an affinity chromatographic step results in an antibody composition sufficiently pure for pharmaceutical uses. The methods described herein comprise pH viral reduction/inactivation, ultrafiltration/diafiltration, affinity chromatography, preferably Protein A affinity, ion exchange chromatography, and hydrophobic chromatography. Further, the present invention is directed toward pharmaceutical compositions comprising one or more antibodies of the present invention.

Claims

1. A process for purifying adalimumab from a fermentation harvest of a Chinese Hamster Ovary (CHO) cell culture expressing said adalimumab, said process comprising:

a) binding adalimumab from said fermentation harvest to a Protein A resin,

b) eluting the bound adalimumab at an elution pH of 3.6-4, and

c) incubating the eluted adalimumab for 1 to 3 hours.

2. The process of claim 1, wherein said eluting is at an elution pH is 3.6.

3. The process of claim 1, wherein said eluting is at an elution pH of 3.7.

4. The process of claim 1, wherein said eluting is at an elution pH of 3.8.

5. The process of claim 1, wherein said eluting is at an elution pH of 4.

6. The process of claim 1, further comprising after said incubating, subjecting adalimumab to one or more further chromatographic separations, wherein said one or more further chromatographic separations comprise an ion exchange chromatography, a hydrophobic interactive chromatography or a combination thereof.

7. The process of claim 1, further comprising subjecting adalimumab to an ion exchange chromatography after said incubating.

8. The process of claim 7, wherein said ion exchange chromatography is cation exchange chromatography.

9. The process of claim 7, wherein said ion exchange chromatography is anion exchange chromatography.

10. The process of claim 1, further comprising subjecting adalimumab to a hydrophobic interaction chromatography after said incubating.

11. The process of claim 1, further comprising determining the monomer level of adalimumab purified according to said process.

12. The process of claim 1, wherein the adalimumab monomer level after said incubating is greater than 97%.

13. The process of claim 1, wherein the purified adalimumab is pharmaceutical grade.