Protein purification
DC CAFCFirst Claim
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1. 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 to a cation exchange material with an equilibration buffer at a first conductivity;
(b) washing the cation exchange material with a wash buffer, wherein the conductivity of the wash buffer increases from a second conductivity that is higher than the first conductivity to a third conductivity during the washing;
(c) passing a fixed volume of wash buffer at the third conductivity over the cation exchange material; and
(d) eluting the antibody from the cation exchange material with an elution buffer at a fourth conductivity that is higher than the third conductivity.
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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.
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14 Claims
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1. A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises the following steps performed sequentially:
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(a) binding the antibody to a cation exchange material with an equilibration buffer at a first conductivity; (b) washing the cation exchange material with a wash buffer, wherein the conductivity of the wash buffer increases from a second conductivity that is higher than the first conductivity to a third conductivity during the washing; (c) passing a fixed volume of wash buffer at the third conductivity over the cation exchange material; and (d) eluting the antibody from the cation exchange material with an elution buffer at a fourth conductivity that is higher than the third conductivity. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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