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US 11,952,399 B2Granted
Methods for purifying proteins
AssigneeAmgen Inc.
InventorsAshish Sharma, Balakumar Thangaraj
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Abstract
The invention pertains to methods of purifying fusion proteins, in particular TNFR:Fc fusion proteins. Methods disclosed herein can be used to produce highly pure TNFR:Fc fusion proteins (e.g., etanercept) having a biological activity by removing hard to separate product related impurities such as clipped and/or mis-fold/aggregated TNFR:Fc fusion proteins.
Claims21
A method for purifying a tumor necrosis factor receptor (TNFR):Fc fusion protein, comprising:
- (a) subjecting a sample comprising the TNFR:Fc fusion protein and at least one contaminant to a chromatography medium under conditions whereby the TNFR:Fc fusion protein binds to the chromatography medium,
- (b) eluting the TNFR:Fc fusion protein from the chromatography medium, and
- (c) filtering the eluate from (b) through
- (i) one or more adsorptive prefilters;
- or
- (ii) one or more charged or surface modified microporous membranes;
- and
- (d) filtering the eluate from (c) through one or more viral filtration membranes to obtain the purified TNFR:Fc fusion protein, wherein the chromatography medium (i) is a hydrophobic interaction chromatography (HIC) medium or (ii) comprises a ligand comprising a hydrophobic group and an ionic group.
5 more elements
Dependencies are inferred from the claim text.