FIELD: biotechnology.
SUBSTANCE: described is a method of purifying the fusion protein TNFR: Fc from a mixture of proteins containing a TNFR: Fc fusion protein and at least one HCP impurity. Said method involves the following steps: a) obtaining from a suitable mammalian expression system a protein mixture containing a TNFR: Fc fusion protein and host cell host (HCP) impurities which contain at least one proteolytic enzyme; b) applying said mixture of proteins to a protein A affinity chromatography column; c) eluting the TNFR: Fc fusion protein from the protein A affinity chromatography step from step (b), d) applying a mixture of proteins obtained at step (c) to a column for hydrophobic interaction chromatography; e) eluting the TNFR: Fc fusion protein from the hydrophobic interaction chromatography column from step (d), f) applying a mixture of proteins obtained at step (e) to an anion-exchange chromatography column; g) eluting the TNFR: Fc fusion protein from the column for anion-exchange chromatography from step (f), h) applying a mixture of proteins obtained at step (g) to a chromatography column with mixed mode; i) elution of the TNFR: Fc fusion protein in the flow mode from the chromatography column with mixed mode from step (h).
EFFECT: invention extends the range of methods for purifying the TNFR: Fc fusion protein.
20 cl, 3 dwg, 2 tbl, 6 ex
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Authors
Dates
2019-08-28—Published
2015-06-13—Filed