Wyeth

Genetics Institute, Inc.

Shujun Sun

Brian D Kelley

Christopher Gallo

Paul Brown

James Edward Booth

Ranganathan Godavarti

Suresh Vunnum

Mary Beth Switzer

Tianning Yu

William Barry Foster

Jonathan L Coffman

Timothy Iskra

Jonathan Lee Coffman

Bonnie J Germain

Yin Luo

Jeffrey J Robinson

Priscilla Jennings

Tim Iskra

Jon Coffman

William B Foster

Brian Kelley

This invention relates to the application of hydroxyapatite chromatography to the purification of at least one antibody from a preparation containing high molecular weight aggregates. Further, this invention relates to an integration of ceramic hydroxyapatite chromatography into a combination chromatographic protocol for the removal of high molecular weight aggregates from an antibody preparation.

Removal of high molecular weight aggregates using hydroxyapatite chromatography

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

The invention relates to methods for isolating a product and/or reducing turbidity and/or impurities from a load fluid comprising the product and one or more impurities by passing the load fluid through a medium, followed by at least one wash solution comprising arginine, and collecting the product using an elution solution. The invention further relates to a product prepared using a method as described herein.

Arginine wash in protein purification using affinity chromatography

The present invention provides a method of removing product-related inactive or partially active species, high molecular weight aggregates, as well as other process-related impurities from preparations of acidic proteins by using ceramic hydroxyapatite chromatography.

Purification of acidic proteins using ceramic hydroxyapatite chromatography

The invention relates to methods for isolating a product and/or reducing turbidity and/or impurities from a load fluid comprising the product and one or more impurities by passing the load fluid through a medium, followed by at least one wash solution comprising an arginine derivative, and collecting the product using an elution solution. The invention further relates to a product prepared using a method as described herein.

Arginine derivative wash in protein purification using affinity chromatography

The present invention provides a process for dissociating Fc-containing molecules from complexes of Protein A/Fc-containing molecules or mixtures containing Fc-containing molecules and Protein A. The association, e.g., by hydrophobic interactions, between the Fc-containing molecules and Protein A can be reduced or inhibited by raising the pH of dissociation. The pH of dissociation can be raised by addition of agents capable of inhibiting hydrophobic interactions, including buffers containing arginine and/or ethylene glycol, to the mixture, either prior to adding the mixture to the column chromatography substrate, after adding the mixture to the column chromatography substrate, or both prior to and after adding the mixture to the column chromatography substrate. Separation of Fc-containing molecules from Protein A can be performed on a number of different column chromatographic substrates, including column chromatographic substrates contained in Q columns (Q column chromatography substrate), HIC columns (hydrophobic interaction column chromatography substrate), and IMAC columns (metal chelate column chromatography substrate).

Methods for purifying Fc-containing proteins

The present invention provides a process for isolating and purifying highly anionic target proteins, for example, sulfated proteins. Sulfated proteins have five (5) or more, sulfations. In a preferred embodiment, the sulfated protein has six (6) sulfations, e.g., six sulfations on N-terminal tyrosine residues, as embodied in PSGL-1 (P-selectin glycoprotein ligand).

Methods for purifying highly anionic proteins

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Brentwood, NH, United States of America

Shujun Sun