ABSTRACT
Hemophilia B is a genetic disorder due to deficiency or complete absence of factor IX coagulation factor. Treatment of choice for these patients is use of factor IX concentrates. Therefore, purification of plasma proteins and separation of factor IX have been major objectives for scientists involved in this field. In this respect, purification procedure using ion exchange chromatography is widely used, but in the past decade affinity chromatography was also introduced. The objective of the present study has been to apply both techniques for the purification of factor IX and compare the quality and yield of the product. For the purification procedure, chromatography columns [XK-16], containing DEAE sepharose and Heparin sepharose were used. Factor IX coagulation activity was measured using a one-stage coagulation assay and factor IX antigen was quantified using ELISA technique. The specific activity and relative increase in purity of factor IX was calculated and it was demonstrated that specific activity improved from 3.1 IU/mg using DEAE ion exchange to 29 IU/mg when affinity chromatography was added and purity was increased from 155 to 1450 respectively. The present study demonstrates that addition of an affinity chromatography step using heparin sepharose is a major improvement in the purification of factor IX, where both specific activity and purity are increased considerably