Production of highly purified clotting factor IX by a combination of different chromatographic methods

A highly enriched preparation of human clotting factor IX was produced by a combination of adsorption chromatography, hydrophobic interaction chromato graphy and heparin affinity chromatography. The introduction of adsorption chromatography with a hydroxyaminopropyl support allows the capture step to be carried out directly from the cryoprecipitate-depleted plasma with a ch romatographic column in flow-through mode. This replaces the batch procedur e used until now. The other two chromatographic steps are designed in such a way that the eluate from the preceding step can be directly applied, with out any intermediate treatment of the sample. This cuts the period of time required for the process by almost 50%, and increases the yield considerabl y. The isolated factor IX contains practically no contaminants and has a sp ecific activity over 200 IU/mg of protein. (C) 1999 Elsevier Science B.V. A ll rights reserved.