METHOD FOR OBTAINING TNFR-Fc FUSION PROTEIN WITH ASSIGNED CONTENT OF IMPURITIES Russian patent published in 2018 - IPC C07K19/00 C07K14/705 C07K16/00 

FIELD: biotechnology.

SUBSTANCE: discloses a method for purifying a TNFR-Fc fusion protein from impurities to produce a protein with a predetermined fraction of a hydrophobic chromatographic peak 3 comprising: (a) introducing a sample containing a liquid mixture of a TNFR-Fc fusion protein derived from mammalian cells partially purified by affinity chromatography, anion exchange chromatography, or both, in a column filled with a hydrophobic interaction hydrophobic interaction (HIC) media containing an aromatic functional group previously equilibrated with a equilibration (EQ) buffer containing sodium chloride in a concentration of 1 M to 1.4 M or ammonium sulfate at a concentration of 0.45 M to 0.55 M; and (b) collecting the eluate by eluting the protein with an elution buffer containing sodium chloride or ammonium sulfate at the same concentration as the equilibration buffer, where a predetermined proportion of the hydrophobic chromatographic peak 3 is from 9 % to 18 %. Invention also relates to a method for correcting the proportion of a hydrophobic chromatographic peak 3 in an etanercept, comprising: preliminary equilibration of the medium for chromatography of hydrophobic interaction with an equilibration buffer containing sodium chloride in a concentration of 1.0 M to 1.5 M or ammonium sulfate at a concentration of 0.45 M to 0.55 M; and introducing a sample containing the TNFR-Fc fusion protein prepared in this way, it is preferable that the concentration of salts in it be the same as in a pre-equilibrated medium for hydrophobic interaction chromatography, where the proportion of the hydrophobic chromatographic peak 3 is adjusted downward as compared to the sample introduced, and wherein the proportion of the hydrophobic chromatographic peak 3 in the sample in which the proportion of the hydrophobic chromatographic peak 3 exceeds 20 % is reduced to a level of 2 % to 17 %.

EFFECT: invention may be useful in the manufacture of biological medicaments containing a recombinant protein, such as etanercept obtained from a mammalian cell culture by a genetic recombination technique.

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