Purification of non-glycosylated polypeptides

US9481706B2 · kind B2 · utility

Assignee

• Hoffmann-La Roche Inc. · US

Inventors

• Roberto Falkenstein · Munich, DE
• Birgit Weydanz · Penzberg, DE
• Nicole Fuehrler · Schlehdorf, DE
• Claudia Giessel · Annaberg-Buchholz, DE
• Sybille Gabel · Peiting, DE
• Adelbert Grossmann · Murnau am Staffelsee, DE
• Friederike Hesse · Munich, DE
• Marc Pompiati · Penzberg, DE
• Andreas Schaubmar · Penzberg, DE
• Brigitte Kraemer · Penzberg, DE

Key dates

• Filing date: Apr 7, 2014
• Grant date: Nov 1, 2016
• Expiry date: Apr 7, 2034

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K1/36
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombinantly produced in a prokaryotic cell, wherein the method comprises three chromatography steps of which the first chromatography step selected from i) hydrophobic charge induction chromatography, or ii) hydrophobic interaction chromatography, or iii) affinity chromatography, or iv) ion exchange chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatography, or iii) hydroxylapatite chromatography, or iv) hydrophobic interaction chromatography, and the a third chromatography step is selected from i) hydrophobic charge induction chromatography, or ii) anion exchange chromatography, or iii) cation exchange chromatography, or iv) hydrophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of interacting with metal ligands, the second chromatography step is not a hydroxylapatite chromatography step in case of polypeptides with an isoelectric point below 6.0, and the third chromatography step can be performed …