膜径向离子交换色谱分离凝血酶原复合物  

Purification of Prothrombin in Nitschmann Fraction Ⅲ by Membrane Radial Column Ion-Exchange Liquid Chromatography

作　　者：孙涛[1] 刘一平 卜凤荣[1] 陈戈[2] 温美娟[2] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850 [2]北京科技大学化学系,北京100083

出　　处：《色谱》2000年第4期350-353,共4页Chinese Journal of Chromatography

摘　　要：利用径向离子交换色谱法分离纯化了Nitschmann组分Ⅲ中的凝血酶原复合物 ( prothrombincomplexconcentrate ,PCC) ,流动相为pH 7.5的Tris HCl缓冲液 ,色谱柱为XK 1 6DEAEfastflowSepharose ( 0 .8cmi.d .× 5cm)及膜DEAE径向色谱柱 ( 3.0cmi.d .× 5.8cm)。通过改变不同上样流速及洗脱流速 ,研究了流速对所分离的凝血酶原复合物的蛋白质量浓度及凝固活性的影响 ,为今后在血浆蛋白分离纯化中进一步推广使用径向色谱技术和放大实验提供了依据。A method is described for the purification of human prothrombin complex concentrate (PCC) from Nitschmann fraction Ⅲ by membrane radial column ion exchange chromatography,which allows large sample volumes to be processed at low operation pressure. The Nitschmann fraction Ⅲ(15 g) was mixed with 1 000 mL 0.06 mol/L Tris HCl(pH 7.5).The centrifuged supernatant(10 000 r/min,15 min,20 ℃)was applied onto a DEAE ion exchange liquid chromatographic column(XK 16 DEAE fast flow Sepharose and DEAE membrane radical column chromatography) with almost the same excellent separation efficiency. The parameters of sample flow rate, elution flow rate and sample capacity were optimized.

关 键 词：Nitschmann组分Ⅲ 凝血酶原复合物 纯化 分离 

分 类 号：Q556.9[生物学—生物化学] R973.2[医药卫生—药品]