凝血因子 Ⅸ复合物制备工艺的优化及性质研究

Technology Optimization and Properties Study of Factor Ⅸ Complex Concentrates

目的 用国产凝胶制备高质量的凝血因子 复合物。方法 以新鲜冰冻血浆为原料 ,经过国产DEAE-琼脂糖快流速阴离子交换层析及 Ca3(PO4 ) 2 吸附法制备凝血因子 复合物。对制备获得的凝血因子 复合物进行凝血活性测定 ,高分子量杂蛋白定量检测以及 SDS- PAGE、免疫电泳 (IE)、交叉免疫电泳 (CIE)等性质分析 ,并且与用进口 DEAE- Sepharose CL- 6 B制备获得的凝血因子 复合物相比较。结果 两步的活性回收率分别为93.6 7± 2 .5 6 % (n=6 )和 88.71± 2 .39% (n=6 )。凝血因子 (F ) :C比活为 7.36± 0 .96 U/ mg(n=6 )。性质分析结果显示 ,本工艺制备的凝血因子 复合物不仅充分保留了有效成分 ,而且制品的纯度和血栓安全性有明显提高。结论 国产凝胶完全能取代进口凝胶用于制备高质量、低成本的凝血因子

Objective To produce high quality factor Ⅸ complex concentrates with domestic ion exchanger. Methods A human solvent detergent (S/D) treated factor Ⅸ complex concentrates has been produced from fresh frozen plasma using domestic DEAE Sepharose fast flow ion exchange chromatagraphy and Ca 3(PO 4) 2 adsorption. The properties of this new concentrates were compared with those of the concentrates obtained by foreign DEAE Sapharose CL 6B. Results The recovery of activity was 93.67±2.56%( n =6) and 88.71±2( n =6),respectively. The specific activity of the concentrates was 7.36±0.96 U Factor Ⅸ: C/mg protein ( n =6).This new concentrates retained all the useful components with higher purity and lower thrombogenicity. Conclusion Domestic ion exchanger could replace foreign ion exchanger in producing high quality factor Ⅸ complex concentrates.