Stability of Activated Recombinant Human Factor VII (rFVIIa) after 28 Days Storage at 24° C and 43° C

The stability of rFVIIa (Novo Seven) was evaluated over a 28 day period. Lyophilized recombinant factor VIIa (1.2 mg/vial configuration) powder was tested at recommended storage conditions of 4°C or accelerated stability testing conditions of 24°C and 65% ± 5% relative humidity (RH) and 43°C plus 65% ± 5% RH. No significant change was noted in the appearance of any of the rFVIIa samples based on turbidity and visual appearance of the lyophilized cake and reconstituted liquid product. rFVIIa activity was determined by bioassay using a modified prothrombin time (PT). Chemical integrity of the rFVIIa product was determined by reverse-phase HPLC and by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). No significant changes were observed in the activity of any of the 28 day-stored samples based on the modified PT as compared to the initial baseline 4°C samples. Reverse Phase-HPLC detected a single primary peak that accounted for 97% of the material. No significant degradation of this peak was observed in any of the 28 day-stored samples compared to the initial baseline 4°C samples. Non-reduced SDS-PAGE revealed a 50,000 molecular weight band and reduced SDS-PAGE showed 30,000 and 20,000 molecular weight bands. No significant changes were observed in the protein bands in any of the 28 day-stored samples analyzed by reduced and non-reduced SDSPAGE as compared to the initial baseline 4°C samples. Taken together, these data suggest that in vitro rFVIIa activity and chemical integrity of the lyophilized powder remain intact for at least 28 days even at 43°C with 65% relative humidity.