色谱2012，Vol.30Issue(6)：618-623,6.DOI:10.3724/SP.J.1123.2012.01036
超大孔离子交换制备色谱分离纯化高活性凝血因子Ⅷ
Separation of coagulation factor Ⅷ with high activity using gigaporous anion exchange chromatography
摘要
Abstract
A purification process to obtain coagulation factor Vffl (FVI) with high activity from human plasma was established. Based on the analysis of the size ratio between FVi 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 substrate , 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 FVI product with high activity. The yield of FVM in superporous chromatography was about five times of commercially agarose chromatography and the specific activity was up to 154 IU/mg protein. Furthermore, we studied the regeneration process of the superporous medium, washing the column with 5 column 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.关键词
超大孔离子交换制备色谱/凝血因子Ⅷ/血浆蛋白/分离纯化Key words
gigaporous preparative ion exchange chromatography/ coagulation factor VI/ plasma protein/ separation and purification分类
化学化工引用本文复制引用
康丽梅,张焱,罗坚,李由,周月芳,余蓉,苏志国..超大孔离子交换制备色谱分离纯化高活性凝血因子Ⅷ[J].色谱,2012,30(6):618-623,6.基金项目
国家高技术研究发展计划("863"计划)(No.2007AA100506)和国家自然科学基金项目(Nos.20806052,20820102036). （"863"计划）
