摘要
目的采用自制的DEAE Bio-SepFF和肝素Bio-Sep FF介质,从人血浆中快速分离纯化凝血因子Ⅸ(FⅨ)。方法低温离心去除冷沉淀后的人血浆,在不同淋洗条件下,经过两步弱阴离子交换和一步亲和层析分离纯化FⅨ,用活性检测试剂盒检测各组分凝血因子的活性,Bradford法测定蛋白浓度,聚丙烯酰胺凝胶电泳分析样品的纯度。结果经过三步层析分离,得到的FⅨ比活达到99.40IU/mg,纯化倍数为3823倍,回收率约为30%,纯度较高。结论采用该方法和介质,可从人血浆中纯化FⅨ,且效果较好。
Objective To rapidly separate and purify blood coagulation factor Ⅸ （FⅨ） from human plasma by using self-made DEAE Bio-Sep FF and heparin Bio-Sep FF chromatographic media. Methods Purify FⅨ from human plasma, after removal of cryoprecipitate by centrifugation at low temperature, by two steps of weak anion exchange and one step of affinity chromatography. Determine the FⅨ in various fractions for activity by activity determination kit, for protein content by Bradford method, and for purity by PAGE. Results After three steps of chromatography, the specific activity, purification factor and recovery of FⅨ were 99. 40%, 3823 and 30% respectively. Little foreign protein was observed on PAGE profile, indicating high purity of purified FⅨ. Conclusion The method and media used in this paper were suitable for purification of FⅨ from human plasma.
出处
《中国生物制品学杂志》
CAS
CSCD
2008年第10期906-908,共3页
Chinese Journal of Biologicals
关键词
弱阴离子交换层析
肝素亲和层析
凝血因子Ⅸ
分离
纯化
Weak anion exchange chromatography
Heparin affinity chromatography
Blood coagulation Factor Ⅸ （FⅨ）
Separation
Purification
