Patent ID: 9556258
Date: 2017-01-31
CPC Classifications: C07K

Claim:
1. A method of purifying a p75 TNFR:Fc-fusion protein from one or more impurites in a sample, comprising the steps of: a) providing a sample comprising the Fc-fusion protein produced in a eukaryotic expression system; b) binding the Fc-fusion protein present in the sample to a Protein A affinity chromatography resin; c) eluting the Fc-fusion protein from the Protein A resin, by applying to the Protein A resin, an elution buffer gradient of from 50 mM Citrate pH 5.0 to 100 mM Citrate pH 4.0, wherein the eluted product provides a second sample; d) binding the second sample to a cation exchange (CEX) chromatography resin; e) eluting the second sample from the CEX resin by applying to the CEX resin, a linear elution buffer gradient of from 25 mM sodium phosphate, 25 mM arginine pH 4.0, 0.25 M NaCl to 25 mM sodium phosphate, 25 mM arginine pH 4.0, 0.65 M NaCl, wherein the eluted product provides a third sample; f) binding the third sample to an anion exchange (AEX) chromatography resin; and g) eluting the third sample from the AEX resin by applying, to the AEX resin, a linear elution buffer gradient of from 12.5 mM sodium phosphate, pH 8.0 to 12.5 mM sodium phosphate, 300 mM NaCl, pH 8.0, wherein the eluted product provides a purified p75 TNFR:Fc-fusion protein composition.