Process for the purification of pharmacologically active proteins through cationic exchange chromatography
AssigneeAlfa Wassermann S.p.A.
InventorsLucia Scapol, Giuseppe Claudio Viscomi
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Abstract
There is disclosed a process for the purification of pharmacologically active proteins based on the use of the cationic exchange chromatography on a solid matrix carried out at a more basic pH, i.e. higher, in respect of the pH corresponding to the isoelectric point, pI, of the proteins to be purified, pH at which however said proteins still remain absorbed. Buffer solutions with values of pH and of ionic strength adjusted from time to time to the kind of pharmacologically active protein to be purified are used in order to obtain such a result. The process is mainly addressed to the purification of the interferon and albumin proteins.
Claims7
A process for the purification of the recombinant alpha-2b interferon, rIFNα-2b, that comprises charging a proteic mixture coming from the manufacturing by fermentation of the rIFNα-2b added with a 1M solution of sodium acetate and brought to pH 5.5 with acetic acid, on a column filled with strong cationic exchange resin conditioned at pH 5.5 by means of a 20 mM solution of sodium acetate so that between 6 and 8 mg of protein are present for each ml of stationary phase, in submitting the column to two washing cycles, first with a buffer solution at pH 6.1 at a concentration between 5 and 15 mM, then with the same buffer solution added with 2 mM of potassium chloride and lastly in eluting the pure rIFNα-2b from the columns by using a buffer solution at pH 6.1 at a concentration between 5 and 15 mM containing potassium chloride at a concentration comprised between 15 and 25 mM.
A process for the purification of interferon proteins which comprises carrying out a cationic exchange chromatography on a solid matrix at a more basic pH than the pH corresponding to the isoelectric point pI of the interferon proteins to be purified, at which pH said proteins still stay absorbed, and eluting said proteins by increasing the ionic strength and/or the pH of the eluents to achieve a purification of the interferon proteins without hydrophobic interaction chromatography or anionic exchange chromatography.
Dependencies are inferred from the claim text.