Method of weak partitioning chromatography
AssigneeWyeth
InventorsBrian D. Kelley, James E. Booth, Paul Brown, Jon Coffman, Ranganathan Godavarti, Tim Iskra, Shujun Sun, Mary B. Switzer, Suresh Vunnum, Tianning Yu
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Abstract
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.
Claims44
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium to obtain a column effluent, at operating conditions which cause the medium to bind at least about 2.8 mg of product per mL of medium, wherein the medium is selected from the group consisting of a charged ion exchange medium, a hydrophobic interaction chromatography resin, and an immobilized metal affinity chromatography resin;
- and
- recovering the purified product in the column effluent.
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium to obtain a column effluent, at operating conditions defined by a partition coefficient of at least about 0.1;
- and
- recovering the purified product in the column effluent.
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium to obtain a column effluent, at operating conditions which cause the medium to bind about 2.8 to about 13 mg of product per mL of medium;
- and
- recovering the purified product in the column effluent.
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium at operating conditions which cause the medium to bind at least about 1 mg of product per mL of medium, wherein the medium is selected from the group consisting of a hydrophobic interaction chromatography resin and an immobilized metal affinity chromatography resin;
- and
- recovering the purified product from column effluent.
A method of recovering a purified antibody from a load fluid, comprising the steps of:
- passing the load fluid through a charged anion exchange medium to obtain a column effluent, at operating conditions which cause the medium to bind at least about 2.8 mg of antibody per mL of medium, wherein the operating conditions comprise a pH of about 7.5 to about 8.2 and a chloride concentration of about 12 to about 55 mM, and wherein the load fluid is a product-containing fluid eluted from a Protein A column;
- and
- recovering the purified antibody in the column effluent.
A method of recovering a purified antibody from a load fluid, comprising the steps of:
- passing the load fluid through a charged anion exchange medium to obtain a column effluent, at operating conditions defined by a partition coefficient of at least about 1.0;
- wherein the operating conditions comprise a pH of about 7.5 to about 8.2 and a chloride concentration of about 12 to about 55 mM, and wherein the load fluid is a product-containing fluid eluted from a Protein A column;
- and
- recovering the purified antibody in the column effluent during the load cycle and any essentially isocratic wash.
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium to obtain a column effluent, at operating conditions which cause the medium to bind at least about 1 mg of product per mL of medium;
- and
- recovering at least about 98% of the product in the column effluent.
A method of recovering a purified product from a load fluid, comprising the steps of:
- passing the load fluid through a medium to obtain a column effluent, at operating conditions which cause the medium to bind at least about 3.0 mg of product per mL of medium;
- and
- recovering at least about 93% of the product in the column effluent.
Dependencies are inferred from the claim text.