Margolis J, Gallovich C M, Rhoades P
Vox Sang. 1984;46(6):341-8. doi: 10.1111/j.1423-0410.1984.tb00097.x.
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 A 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 micron 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 unit of factor VIII per mg protein (10 units mg fibrinogen) with a recovery of 250 units/kg plasma. The CPG stage is non-destructive, yielding more than 90% of the input factor VIII. In 1980-1983, more than 3 X 10(6) units have been 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.
详细描述了一种大规模生产“高纯度”因子VIII的简单方法。由洗涤后的冷沉淀制备的粗浓缩物，以20 - 30毫升孔径为500埃的可控孔径玻璃（CPG）与1克输入蛋白质的比例进行处理。将浆液倒入分离柱中，收集流出的纯化浓缩物。通过洗涤液置换空体积中剩余的因子VIII。通过0.2微米的膜过滤器后，将产品分装并冻干。将操作pH维持在6.5 - 6.7并添加合成氨基酸可提高产量和溶解度。目前的浓缩物每毫克蛋白质含有1单位因子VIII（每毫克纤维蛋白原含10单位），每千克血浆的回收率为250单位。CPG阶段不会破坏因子VIII，输入的因子VIII产率超过90%。1980 - 1983年期间，新南威尔士州使用了超过3×10⁶单位，主要用于外科手术患者的大量覆盖。与联邦血清实验室合作，计划扩大生产以供澳大利亚其他州使用。
