Protein purification
AssigneeGenentech, Inc.
InventorsCarol D. Basey, Greg S. Blank
Life of this patent
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Abstract
A method for purifying a polypeptide by ion exchange chromatography is described which involves changing the conductivity and/or pH of buffers in order to resolve a polypeptide of interest from one or more contaminants.
Claims22
A method for purifying a polypeptide from a composition comprising the polypeptide and a deamidated variant thereof, which method comprises the following steps performed sequentially:
- (a) binding the polypeptide and deamidated variant to an ion exchange material using a loading buffer, wherein the loading buffer is at a first conductivity and pH;
- (b) washing the ion exchange material with an intermediate buffer at a second conductivity and/or pH so as to elute the deamidated variant from the ion exchange material;
- (c) washing the ion exchange material with a wash buffer which is at a third conductivity and/or pH, wherein the change in conductivity and/or pH from the intermediate buffer to the wash buffer is in an opposite
- direction to the change in conductivity and/or pH from the loading buffer to the intersediate buffer;
- and
- (d) washing the ion exchange material with an elution buffer at a fourth conductivity and/or pH so as to elute the polypeptide from the ion exchange material.
A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises the following steps performed sequentially:
- (a) binding the antibody and contaminant to a cation exchange material using a loading buffer, wherein the loading buffer is at a first conductivity and pH;
- (b) washing the cation exchange material with an intermiediate buffer at a second conductivity and/or pH which is greater than that of the loading buffer so as to elute the contaminant from the cation exchange material;
- (c) washing the cation exchange material with a wash buffer which is at a third conductivity and/or pH which is less than that of the intermediate buffer;
- and
- (d) washing the cation exchange material with an elution buffer at a fourth conductivity and/or pH which is greater than that of the intermediate buffer so as to elute the antibody from the cation exchange material.
A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises the following steps performed sequentially:
- (a) binding the antibody and contaminant to an ion exchange material using a loading buffer, wherein the loading buffer is at a first conductivity and pH;
- (b) washing the ion exchange material with an intermediate buffer at a second conductivity and/or pH so as to elect the contaminant from the ion exchange material;
- (c) washing the ion exchange material with a wash buffer which is at a third conductivity and/or pH, wherein the change in conductivity and/or pH from the intermediate buffer to the wash buffer is in an opposite direction to the change in conductivity and/or pH from the loading buffer to the intermediate buffer;
- and
- (d) washing the ion exchange material with an elution buffer at a fourth conductivity and/or pH so as to elute the antibody from the ion exchange material.
Dependencies are inferred from the claim text.