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US 8,263,750 B2Granted
Method for purifying a protein using protein-A affinity chromatography using an intermediate wash step
AssigneeAmgen Inc.
InventorsAbhinav A. Shukla, Peter Hinckley
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Abstract
A method for purifying a protein using Protein A chromatography comprising a) absorbing the protein to Protein A immobilized on a solid support; b) removing contaminants by washing the immobilized Protein A containing the absorbed protein with a buffer comprising one or more chaotropic agents in combination with one or more hydrophobic modifiers and having a pH of at least 7.0; and c) eluting the protein from the Protein A immobilized on the solid support.
Claims5
A method for purifying an IgG1 or IgG2 antibody from a solution containing at least one contaminant by Protein A chromatography comprising:
- a) absorbing the IgG1 or IgG2 antibody to Protein A immobilized on a solid support;
- b) removing contaminants by washing the immobilized Protein A containing the absorbed IgG1 or IgG2 antibody with a wash buffer comprising 25 mM Tris, 1M to 3M urea, and an organic solvent selected from 10% (v/v) isopropanol or 10% (w/v) propylene glycol, and wherein said wash buffer has a pH of at least 9.0;
- and
- c) eluting the IgG1 or IgG2 antibody from the Protein A immobilized on the solid support.
1 more element
Dependencies are inferred from the claim text.