Patent ID: 8399632
Filing Date: 2013-03-19
Classification: C07K

Abstract:
1. A method for purification of a recombinant Factor IX (rFIX) composition enriched in monosulfated and/or monophosphorylated rFIX, wherein the method comprises the steps of: a) loading a composition comprising rFIX onto an anion exchange material; b) washing the anion exchange material using a high salt wash buffer having a conductivity of 22±1 mS/cm at 25° C., thereby forming a high salt wash buffer fraction comprising rFIX, wherein less than 30% of the rFIX in the high salt wash buffer fraction is monosulfated and/or monophosphorylated; c) eluting rFIX from the anion exchange material using a gradient of anions in an elution buffer comprising divalent cations; d) collecting the rFIX eluate in fractions; and e) pooling fractions enriched in monosulfated and/or monophosphorylated rFIX, thereby forming a purified rFIX composition enriched in monosulfated and/or monophosphorylated rFIX, wherein at least 30% of the rFIX in the purified rFIX composition are monosulfated and/or monophosphorylated.