DEVELOPMENT AND SMALL-SCALE PRODUCTION OF A SEVERELY HEATED FACTOR-VIII CONCENTRATE

The small-scale production of a severely heated factor VIII concentrat e is described. As starting material for the production 48 units of fr esh frozen plasma from whole-blood donations or 24 units of plasma fro m apheresis are used. During a glycine and sodium chloride fractionati on, contaminating proteins are removed from a pooled extract of single -donor cryoprecipitates. The resulting factor VIII-rich precipitate is redissolved in freeze-drying buffer and this solution is desalted by diafiltration. After filtration, this solution is dispensed into 10 vi als and frozen. The product is freeze dried, and heat treated at 80 de grees C for 72 h. The mean yield of the heat-treated product improved from 160 IU factor VIII per liter plasma at the start of the productio n to 215 IU per liter plasma after a modification of the purification process. The validation of the virus inactivation during freeze-drying and 80 degrees C heat treatment for 72 h showed a reduction of greate r than or equal to 7.6 log PFU/ml for Sindbis and a reduction of great er than or equal to 6.4 log TCID50/ml for HIV-1. The factor VIII conce ntrate was clinically tested in 6 patients. It possessed a normal half -life, showed a high recovery and no side effects. A Dutch license was granted in June 1991 and since then this product has been routinely m anufactured in three Dutch blood banks.