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US 8,044,017 B2Granted
Protein purification
AssigneeGenentech, Inc.
InventorsJefferson C. Emery, Paul J. McDonald, Rhona O'Leary
Life of this patent
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Abstract
A method for purifying a polypeptide by ion exchange chromatography is described in which a gradient wash is used to resolve a polypeptide of interest from one or more contaminants.
Claims25
A method for purifying a polypeptide from a composition comprising the polypeptide and contaminants, which method comprises the sequential steps of:
- (a) loading the composition onto an ion exchange resin with an equilibration buffer having a first salt concentration;
- (b) washing the ion exchange resin with a wash buffer until a predetermined protein concentration is measured in the flowthrough, wherein the salt concentration of the wash buffer increases from an initial, second salt concentration that is greater than the salt concentration of the equilibration buffer, to a final, third salt concentration;
- (c) passing a fixed volume of wash buffer at the final, third salt concentration over the cation exchange resin;
- and
- (d) eluting the polypeptide from the ion exchange resin with elution buffer that has a salt concentration that is greater than the final salt concentration of the wash buffer.
2 more elements
Dependencies are inferred from the claim text.