Effects of Plasma Collection Systems and Processing Parameters on the Quality of Factor IX Concentrate¹

Abstract

Using pilot-scale production of our present factor IX (II and X) concentrate, we have studied the effects of starting plasma source and processing parameter on two in-vitro indicators of product quality - yield and thrombogenic potential. Plasma source did not affect factor IX yield but had a marked effect on thrombogenic potential. Factor IX concentrates produced from plasma derived through centrifugation-based technology showed significantly higher thrombogenic potential than products derived from plasma derived through a filtration-based system. Removal of Cohn fraction I prior to ion-exchange chromatography resulted in a drop in factor IX yield and thrombogenic potential, as did heat treatment to 80 degrees C for 72 h. We conclude that a membrane-filtration-based plasmapheresis system may be the preferred method of plasma procurement for factor IX concentrate production.

Comparison of source plasma quality in three different apheresis protocols

“…The most striking findings were markedly increased PF4 levels in PCS2 plasma. This has not been reported previously but is not surprising, because PCS2 plasma units but not Auto‐C units contain comparatively high platelet counts 1,4,5 . Whether high PF4 levels in plasma units are associated with a higher risk of coagulation activation and thrombosis when transfused in predisposed patients or with lower yields of clotting factors during plasma fractionation remains to be determined in future trials.…”

“…The Turbo Mode program in the Auto‐C increases the rates of blood drawing from 100 to 120 mL/min and returning rates from 130 to 150 mL/min. Despite a close association between source plasma quality and the yield of plasma proteins during plasma fractionation, studies directly comparing the quality of source plasma obtained with the PCS2 and the Auto‐C Normal Mode and Turbo Mode were carried out in the late 1980s for a small set of coagulation factors 1,2 and in 2003 for residual cells and protein composition at a single apheresis machine 3‐5 . We compared the levels of coagulation factors and activation markers in plasma made by the PCS2 and the Auto‐C Normal Mode and Turbo Mode .…”

Heparin/heparan sulfate anticoagulant glycosaminoglycans in human plasma of healthy donors: preliminary study on a small group of recruits

“…Plasmatic HSGAGs were heterogeneous for molecular weight. High-molecularweight HSGAGs were associated with cryoprecipitate and fraction I; that is, those fractions containing proteases involved in the coagulation cascade [19][20][21]. Fraction IV-1, containing antithrombin III [22], showed HSGAGs of smaller size (12.0 kDa).…”

“…HSGAGs endowed with anticoagulant activity therefore did not appear randomly associated with plasma proteins. Most coagulation factors and proteins involved in the control of hemostasis are concentrated in cryoprecipitate (fibrinogen, von Willebrand/factor VIII complex) and in fractions I (factor IX, factor X and prothrombin) and IV-1 (antithrombin III and protein C) [19][20][21][22][23]. These are the fractions where the highest amount of anticoagulant HSGAGs was recovered, thus suggesting a specific association of HSGAGs with proteins involved in hemostatic balance.…”

Factor IX Concentrates for Clinical Use

“…In another study, preparations from plasma obtained from centrifuged donor blood were associated with a higher thrombogenic potential than those made from plasma obtained by pheresis. 41 Improving the plasma as a starting material is particularly applicable to small-scale production, and it emphasizes the importance of decreasing proteolysis prior to fractionation. The twofold excess of antigenic over functional levels of the vitamin K-dependent proteins in the crude concentrates 12 is best explained by proteolytic inactivation during processing.…”