一步离子交换层析制备人血管性血友病因子/人凝血因子Ⅷ蛋白液  

Preparation of human von Willebrand factor/human coagulation factor Ⅷ solution by one-step ion exchange chromatography

作　　者：陈新朋 匡青芬 罗二华 梁小明 刘敏亮 龚佳莉 CHEN Xinpeng;KUANG Qingfen;LUO Erhua;LIANG Xiaoming;LIU Minliang;GONG Jiali(Boya BioPharmaceutical Co Ltd,Enterprise Technology Center of China,Blood Product Research Lab of Jiangxi,Blood Product Engineering Research Center of Jiangxi,Enterprise Technology Center of Jiangxi,Fuzhou 344000,China)

机构地区：[1]博雅生物制药集团股份有限公司,国家企业技术中心,江西省血液制品重点实验室,江西省血液制品工程研究中心,江西省企业技术中心,江西抚州344000

出　　处：《中国输血杂志》2020年第11期1147-1150,共4页Chinese Journal of Blood Transfusion

基　　金：江西省重大科技研发专项(20173ABC28010);江西省省级优势科技创新团队项(20181BCB 24017)。

摘　　要：目的研究通过一步离子交换层析法,制备vWF∶Rco/FⅧ∶C≈1的人血管性血友病因子/人凝血因子Ⅷ蛋白液。方法选用Toyopearl? DEAE-650填料,取同一批次的人血管性血友病因子(vWF)上柱前液,研究不同上样量、不同洗脱液盐浓度对洗脱液中人血管性血友病因子抗原(vWF∶Ag)、人凝血因子Ⅷ效价(FⅧ∶C)、人血管性血友病因子效价(vWF∶Rco)的影响。结果从得率考虑,上样量最佳为4-5倍柱体积;洗脱液盐浓度0.42 mol/L时,vWF∶Rco/FⅧ∶C≈1∶1,vWF回收率≈60%,为最佳配方。结论在本试验层析条件前提下,调节洗脱液中盐浓度0.42 mol/L,可获得vWF∶Rco/FⅧ∶C≈1∶1的vWF/FⅧ蛋白液。Objective To prepare human von Willebrand factor/human coagulation factor Ⅷ complex protein solution with C≈1(vWF∶Rco/FⅧ∶C≈1)by one-step ion exchange chromatography.Methods The same batch of vWF precolumn solution was taken with Toyopearl~? DEAE-650 fillerto study the effects of different loading volume and eluent salt concentration on the titer of human vWF antigen(vWF∶Ag), human coagulation factor Ⅷ(FⅧ∶C) and human vWF(vWF∶Rco).Results Considering the yield, the optimum loading volume was 4 to 5 times of column volume;and the best formulation was vWF∶Rco/FⅧ∶C≈1∶1, and vWF recovery≈60% as the eluate concentration was 0.42 mmol/L. Conclusion Under the condition of chromatography in this experiment, vWF∶Rco/FⅧ∶C≈1∶1 protein solution can be obtained by adjusting the salt concentration at 0.42 mol/L.

关 键 词：人血管性血友病因子 人凝血因子Ⅷ 效价 

分 类 号：R457.1[医药卫生—治疗学]