A Process for Preparation of ‘High‐Purity’ Factor VIII by Controlled Pore Glass Treatment

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Abstract
A simple process for large-scale manufacture of high-purity factor VIII is described in detail. A crude concentrate prepared from washed cryo is treated with controlled pore glass (CPG, 500 .ANG. pore diameter) in proportion of 20-30 ml of CPG to 1 g input of protein. The slurry is poured into a separation column and the effluent purified concentrate collected. The remaining factor VIII in the void volume is displaced by a wash solution. After passage through a 0.2 .mu.m membrane filter the product is dispensed and lyophilized. Maintaining the operating pH at 6.5-6.7 and adding synthetic amino acids improved the yield and solubility. The current concentrate contains 1 U of factor VIII/mg protein (10 U mg fibrinogen) with a recovery of 250 U/kg plasma. The CPG stage is non-destructive, yielding > 90% of the input factor VIII. More than 3 .times. 106 U were used in New South Wales, mostly for massive cover in surgical patients. In collaboration with the Commonwealth Serum Laboratories, it is intended to expand production for use in other Australian States.