Isolation and purification of antibodies using protein A affinity chromatography
AssigneeAbbVie Inc.
InventorsRobert K. Hickman, Qing Huang, Cheryl L. Weed, Scott T. Ennis, Barbara Perilli-Palmer, Min Wan
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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.
Claims18
A method for producing a host cell-protein (HCP) reduced anti-IL-12 antibody, or antigen binding portion thereof, preparation from a sample mixture comprising an anti-IL-12 antibody, or antigen binding portion thereof, and at least one HCP, said method comprising:
- (a) subjecting said sample mixture to a reduction in pH thus forming a reduced pH primary recovery sample, wherein said reduction in pH is from about 3 to about 4;
- (b) adjusting said reduced pH primary recovery sample to a pH of about 6.0 to about 8 thus forming an adjusted pH primary recovery sample followed by;
- (c) contacting said adjusted pH primary recovery sample to a Protein A affinity chromatography resin, washing said affinity chromatography resin with a buffer comprising 0.5 M NaCl, 20 mM citric acid/Na citrate, at pH 6, and collecting an affinity chromatography sample;
- (d) contacting said affinity chromatography sample to an ion exchange resin and collecting an ion exchange sample;
- (e) contacting said ion exchange sample to a hydrophobic interactive chromatography (HIC) resin and collecting an HIC sample, wherein said HIC sample comprises said HCP-reduced anti-IL-12 antibody, or antigen binding portion thereof, preparation.
Dependencies are inferred from the claim text.