Process for the purification and/or isolation of...

C - Chemistry – Metallurgy – 07 – K

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C07K 14/535 (2006.01) A61K 38/19 (2006.01) C07K 1/22 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2469984

The invention relates to the process for the isolation of biologically active granulocyte colony stimulating factor (G-CSF), which enables the separation of correctly folded biologically active monomeric molecules of G-CSF from the incorrectly folded, biologically inactive monomeric, oligo- or polymeric and also from aggregated molecules of G-CSF by using immobilised metal affinity chromatography. The process of the invention, if desired the whole process, can be advantageously performed under native conditions. The biologically active G-CSF with a purity of greater than 95% is thus obtained. Only two additional chromatographic steps, cationic exchange chromatography and gel filtration, are then preferably applied to remove the traces of impurities. The entire process results in the production of higher yields of G-CSF with a purity of greater than 99%. The described process is particularly suitable for the industrial production of G-CSF.

L'invention concerne un procédé d'isolation de facteur biologiquement actif de stimulation des colonies de granulocytes (G-CSF), qui permet d'établir une séparation entre des molécules monomères biologiquement actives correctement pliées de G-CSF et des molécules oligo- ou polymères biologiquement inactives pliées de façon incorrecte et agglomérées de G-CSF par chromatographie d'affinité pour un métal immobilisé. Le procédé de cette invention, si besoin est l'ensemble du procédé, peut être mis en oeuvre, de façon avantageuse, en conditions natives. Le G-CSF biologiquement actif d'un degré de pureté supérieur à 95 % est ainsi obtenu. Les traces d'impuretés sont ensuite retirées, de préférence, en seulement deux étapes chromatographiques supplémentaires, à savoir la chromatographie d'échange de cations et la filtration sur gel. Le présent procédé permet d'obtenir des rendements supérieurs de G-CSF d'un degré de pureté supérieur à 99 % et convient, en particulier, à la production industrielle de G-CSF.

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