Of Cryopreservation, Where Biological Systems Are Typically Frozen in the Presence of Chemicals like Me 2 So. (conflicts of Interest: None Declared. Source of Funding: Mechanical Engineering Department at Louisiana State University and an Economic Development Assistantship (eda) to Dp.)

156. Cryopreservation of recombinant antihemophilic Factor VIII. Effect of biopolymers. Nelly M. Tsvetkova, Omkar Joshi, Arnaud Desponds, D.Q. Wang, Paul Wu, Bayer HealthCare LLC, Berkeley, CA, USA Recombinant Factor VIII, an essential coagulation factor in the blood, is one of the most complex proteins manufactured today. Currently the only therapy for hemophilia A is life-long administration of Factor VIII. However, due to its limited stability during the manufacturing process, there is a constant need for improving its stability. The aim of this study was to evaluate the stability of recombinant Factor VIII process intermediates in the frozen state during 6 months storage at 4 different subambient temperatures, and to test the effect of several non-crystallizing stabilizers on its activity. To this end, thermal analysis using differential scanning calorimetry (DSC) was used to evaluate the phase behavior and glass transition properties of aqueous solutions of sucrose, trehalose, hydroxyethyl starch (HES) and dextran. DSC data showed that the glass transition temperatures (Tg 0) increased from 55 C in the absence of HES and dextran to 35 C in the presence of these polymers. Based on these results, Factor VIII process intermediates formulated with sucrose, trehalose, HES and dextran were stored at 80 C (below the Tg 0), at 30 C (close to Tg 0), and at 18 C and 14 C (above the Tg 0) for 6 months. The data showed that sucrose, trehalose and HES provided good stability of Factor VIII during storage at 80 and 30 C. Kinetic analysis demonstrated that Factor VIII activity depends on the storage temperature when HES or dextran were not part of the formulation. In contrast, in the presence of these polymers, Factor VIII activity did not depend on the storage temperature. These results are discussed in terms of the importance of the glass transition for long term storage of process intermediates. (Conflicts of interest: None declared. Source of funding: None declared).