Wyeth

Genetics Institute, Inc.

Suresh Vunnum

Brian D Kelley

James Edward Booth

Paul Brown

Shujun Sun

Ranganathan Godavarti

Mary Beth Switzer

Tianning Yu

Timothy Iskra

Jonathan Lee Coffman

Johanna Dalborg

Jon Coffman

Tim Iskra

Anna Petersson

This invention relates to methods of using weak partitioning chromatography for the purification of a product from a load fluid containing one or more impurities. Further, the invention relates to methods of weak partitioning chromatography defined by operating conditions which cause a medium to bind least 1 mg of product per mL of medium, or alternatively, defined by a partition coefficient of at least 0.1.

Method of weak partitioning chromatography

Methods are provided for purification of Factor VIII polypeptides by affinity chromatography and ion exchange chromatography, in which the eluate from the affinity column is diluted with a solution comprising higher salt concentration, or lower non-polar agent concentration than that of the elution solution, prior to passing the diluted solution through the ion exchange column. The affinity matrix may comprise a monoclonal antibody or a peptide ligand. The methods result in improved purification without significant yield loss.

Methods for producing Factor VIII proteins

Methods are provided for purification of Factor VIII polypeptides by immunoaffinity chromatography and ion exchange chromatography, in which the eluate from the immunoaffinity column is diluted with a solution comprising higher salt concentration, or lower non-polar agent concentration than that of the elution solution, prior to passing the diluted solution through the ion exchange column. The methods result in improved purification without significant yield loss.

Methods for producing factor VIII proteins

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Burlington, MA, United States of America

Suresh Vunnum