| abstract |
Method for producing an antibody, or portion thereof antigen-binding, with a reduced level of host cell protein (HCP) from a sample mixture comprising an antibody, or part thereof binding to the antigen, and the minus one HCP, and said method comprises: (a) subjecting said sample mixture to a pH reduction, thereby forming a primary recovery sample in which the pH of said sample mixture is reduced to a pH of 3 to 4 to form said primary recovery sample; (b) adjusting said primary recovery sample to a pH of 6.8 to 8, followed by; (c) contacting said primary recovery sample with a protein A affinity chromatographic resin, washing said affinity chromatographic queen with a buffer selected from the group consisting of: 1) a buffer comprising 0.5 M NaCl , 25 mM sodium acetate, at pH 5; 2) a buffer comprising 0.5 M NaCl, 25 mM sodium acetate, at pH 5.5; and 3) a buffer comprising 0.5 M NaCl, 20 mM citric acid / sodium citrate, at pH 6, and collection of an affinity chromatography sample; (d) contacting said affinity chromatography sample with an ion exchange resin and collecting an ion exchange sample; (e) contacting said ion exchange sample with a hydrophobic interaction chromatography (HIC) resin and collecting an HIC sample, wherein said HIC sample comprises said antibody preparation, or binding portion thereof to the antigen, with a reduced level of HCP. |