Process for producing erythropoietin containing no animal...

Details Process for producing erythropoietin containing no animal... Process for producing erythropoietin containing no animal...

C - Chemistry, Metallurgy

07

K

C07K 14/505 (2006.01) A61K 38/18 (2006.01)

Patent

CA 2220515

A preparation of a protein with erythropoietin activity and obtainable after culturing an erythropoietin-producing host cell is characterised by a) a host- cell proteins content of 100 ppm, b) a host cell DNA content of 10 pg per 83 µg of erythropoietin, and c) a total absence of mammalian proteins not derived from the host cell. The preparation is obtained following serum-free culture using a purification process involving dye chromatography, hydrophobic chromatography on an alkylated or arylated carrier, chromatography on hydroxyapatite, hydrophobic chromatography and anion exchange chromatography.

Une préparation d'une protéine présentant l'activité de l'érythropoïétine et pouvant être obtenue après la mise en culture d'une cellule hôte produisant de l'érythropoïétine se caractérise par: a) une teneur en protéines provenant de la cellule hôte de <= 100 ppm, b) une teneur en ADN provenant de la cellule hôte de <= 10 pg/83 µg d'érythropoïétine, et c) une absence totale de protéines de mammifère ne provenant pas de la cellule hôte. Cette préparation est obtenue après la culture sans sérum selon un procédé de purification impliquant une chromatographie sur colorant, une chromatographie hydrophobe sur un porteur alkylé ou arylé, une chromatographie sur hydroxyapatite, une chromatographie hydrophobe et une chromatographie sur échangeur d'anions.

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