C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
C07K 1/18 (2006.01) C07K 14/745 (2006.01)
Patent
CA 2742246
The invention relates to the purification of different gamma carboxylated forms of a polypeptide using ion exchange chromatography. In particular, the invention provides a method for purifying a polypeptide having a desired content of gamma-carboxyglutamic acid from a sample comprising mixture of species of said polypeptide having different contents of gamma-carboxyglutamic acid, said method comprising the steps of: (a) loading said sample onto an anion exchange chromatography material; (b) eluting said polypeptide using a solution at a pH of less than pH 9.0 comprising at least one salt selected from ammonium acetate, ammonium chloride and sodium acetate; and (c) selecting a fraction obtained from said elution wherein the polypeptides in the fraction have the desired content of gamma-carboxyglutamic acids.
La présente invention concerne la purification de différentes formes gamma carboxylées dun polypeptide au moyen dune chromatographie par échange dions. En particulier, linvention concerne un procédé permettant lépuration dun polypeptide possédant une teneur souhaitable en acide gamma-carboxyglutamique dérivé dun échantillon comportant un mélange despèces dudit polypeptide ayant des teneurs différentes en acide gamma-carboxyglutamique. Ledit procédé comprend les étapes suivantes : (a) le chargement dudit échantillon sur un matériau de chromatographie par échange danions ; (b), lélution dudit polypeptide à laide dune solution à un pH inférieur à pH 9,0 comportant au moins un sel sélectionné parmi lacétate dammonium, le chlorure dammonium et lacétate de sodium ; et (c) la sélection dune fraction obtenue à partir de ladite élution, le polypeptide dans la fraction ayant une teneur souhaitable en acide gamma-carboxyglutamique.
Dimock Stratton Llp
Novo Nordisk Health Care Ag
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