中文摘要：

摘要：建立了一条从人血浆中分离高活性凝血因子Ⅷ（FⅧ）的纯化工艺。綦于FⅧ和介质孔径的尺度比及其对蛋白质活性影响的分析，设计了以超大孔离子交换制备色谱为核心步骤的新型分离纯化工艺。分别进行超大孔离子交换色谱与传统离子交换色谱的条件优化，并对优化工艺所得产品进行了活性检测（底物显色法）和纯度检测（高效凝胶过滤和凝胶电泳）。结果表明，超大孔介质结构不但可以有效地保护蛋白质大分子结构，而且能够大幅度地提高制备色谱的传质速率，从而得到具有高凝血活性的FⅧ产品。FⅧ在超大孔制备色谱过程中的州收率（85％）比传统离子交换制备色谱高4-5倍，产品比活高达154IU／mg。此外，还研究了超大孔介质的再生程序，采用5个柱体积的1mol／LNaOH低流速清洗色谱柱，保证了色谱工艺的稳定性、，本纯化工艺步骤简单，重现性好，易于放大生产。

英文摘要：

A purification process to obtain coagulation factor Ⅷ （FⅧ） with high activity from human plasma was established. Based on the analysis of the size ratio between FⅧ and matrix porous medium and its effect on the protein activity, a novel purification process designed was superporous ion exchange chromatography （ IEC）. The operating conditions of gigaporous and traditional anion exchange chromatography were optimized separately. The chromogenic sub- strate, gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS- PAGE） were used to monitor the bioactivity and purity of the chromatographic products. The results showed that the superporous medium could not only protect structure of macro-protein but also enhance its mass transfer, finally giving FⅧ product with high activity. The yield of FⅧ in superporous chromatography was about five times of commercially agarose chromatogra- phy and the specific activity was up to 154 IU/mg protein. Furthermore, we studied the regen- eration process of the superporous medium, washing the column with 5 colunm volumes of 1mol/L NaOH at a low flow rate, to ensure the chromatographic stability. This purification process is simple, reproducible and suitable for large-scale production.