Method of separating glycosylated and non-glycosylated proteins

C - Chemistry – Metallurgy – 07 – K

Patent

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Details

C07K 1/18 (2006.01) C07K 14/575 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2160703

This invention relates to a method for separating glycosylated proteins from non-glycosylated proteins. The method comprises the following steps: 1) equilibrating the protein containing solution with an equilibrating solution containing at least one organic solvent; 2) adding the equilibrated protein solution onto a chromatographic column containing a packing for the ion-exchange affinity separation of glycosylated and non-glycosylated proteins; 3) eluting the glycosylated proteins from the chromatographic column with a sufficient quantity of a first eluting solution containing at least one organic solvent and with or without at least one salt, wherein the salt concentration and/or organic solvent concentration and/or pH are appropriate for the eluting of glycosylated proteins being substantially free of non-glycosylated proteins; and 4) eluting the non-glycosylated proteins from the chromatographic column with a sufficient quantity of a second eluting solution containing at least one organic solvent and with or without at least one salt, wherein the salt concentration and/or organic solvent concentration and/or pH are appropriate for the eluting of non-glycosylated proteins being substantially free of glycosylated proteins.

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