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US 11,236,126 B2Granted
Methods for enhanced removal of impurities during protein A chromatography
AssigneeGenzyme Corporation
InventorCarl A. Beigie
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Abstract
Provided herein are methods relating to the purification of a polypeptide comprising an Fc region (e.g., an antibody) via protein A chromatography; methods relating to the use of a wash solution comprising a benzoate salt and/or benzyl alcohol during protein A chromatography; and methods of adjusting a harvest using sodium benzoate prior to protein A chromatography.
Claims20
A method of purifying a polypeptide comprising an Fc region, the method comprising the steps of:
- (a) contacting a Protein A chromatography matrix with a sample comprising (i) the polypeptide comprising the Fc region, and (ii) one or more impurities, under a condition that the polypeptide comprising the Fc region binds to Protein A;
- and
- (b) washing the matrix with a wash solution, wherein the wash solution comprises one or both of (i) a benzoate salt at a concentration of about 0.1 M to about 1.0 M and (ii) benzyl alcohol at a concentration of about 0.5% to about 4% volume/volume (v/v), and wherein the wash solution has a pH of about 4.0 to about 10.0.
A method of purifying a polypeptide comprising an Fc region, the method comprising the steps of:
- (A) adjusting a harvest comprising the polypeptide comprising the Fc region to achieve a final concentration of a benzoate salt of about 0.1M and about 0.5M and a pH between about 7.0 and about 9.0 to produce a sample comprising (i) the polypeptide comprising the Fc region, and (ii) one or more impurities;
- and
- (B) contacting the sample with at least one chromatography matrix.
Dependencies are inferred from the claim text.