| abstract |
The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombi-nantly 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 ex-change chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatogra-phy, or iii) hydroxylapatite chromatography, or iv) hydrophobic interac-tion 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) hy-drophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of in-teracting with metal ligands, the second chromatography step is not a hy-droxylapatite chromatography step in case of polypeptides with an iso-electric point below 6.0, and the third chromatography step can be per-formed in flow-through mode with polypeptides having a low or high iso-electric point. |