Methods of reducing contaminants in protein purification
AssigneeEli Lilly and Company
InventorsAmy Lynn HUEBNER, Timothy BLANC
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Abstract
Provided herein are improved methods of purifying an Fc-containing protein from a mixture comprising the Fc-containing protein and one or more host cell protein (HCP). The methods generally involve applying the mixture to a chromatography column comprising a protein A chromatography matrix, and washing the chromatography matrix with a sodium caprylate wash buffer.
Claims56
A method of purifying dulaglutide from a mixture comprising the dulaglutide and one or more host cell protein (HCP), the method comprising the steps of:
- (a) applying the mixture to a chromatography column comprising a Protein A chromatography matrix, under conditions whereby the dulaglutide binds to the chromatography matrix;
- and
- (b) washing the chromatography matrix with a sodium caprylate wash buffer comprising 150-500 mM sodium caprylate.
A method of purifying dulaglutide from a mixture comprising the dulaglutide and one or more host cell protein (HCP), the method comprising the steps of:
- (a) applying the mixture to a chromatography column comprising a Protein A chromatography matrix, under conditions whereby the dulaglutide binds to the chromatography matrix;
- (b) washing the chromatography matrix with about 2 column volumes of a NaCl wash buffer comprising about 50 mM Tris and about 1 M NaCl, with a pH of about 8;
- (c) washing the chromatography matrix with about 2 column volumes of a sodium caprylate wash buffer comprising about 50 mM Tris and about 300 mM sodium caprylate, with a pH of about 8;
- (d) washing the chromatography matrix with a Tris buffer comprising about 50 mM Tris at a pH of about 8.0;
- and
- (e) contacting the chromatography matrix with an elution buffer comprising about 10 mM sodium citrate and having a pH of about 3 to produce an eluate containing dulaglutide;
- wherein the eluate comprises:
- at least 70% of the dulaglutide present in the mixture and no more than 100 ng/mg of HCPs;
- and wherein the one or more HCPs includes cathepsin D.
Dependencies are inferred from the claim text.