猪凝血酶的分离纯化与鉴定  

The Isolation,Purification and Characterization of Thrombin from Porcine Plasma

作　　者：徐长法[1] 王兰仙[1] 刘德富[1] 马晓岚[1] 

机构地区：[1]北京大学生物学系,北京联合大学文理学院

出　　处：《北京联合大学学报》1994年第1期44-49,共6页Journal of Beijing Union University

摘　　要：本文报导以猪血为材料，制备凝血酶，血浆在低离子强度下经等电沉淀得优球蛋白，从该沉淀中采用稀盐溶液提取，氢氧化镁吸附和二氧化碳解吸等步骤获得凝血酶原粗品。后者经脑提取液激活，再经乙醇分级、ＤＥＡＥ－纤维素与磷酸纤维素离子交换层析和ＳｅｐｈａｄｅｘＧ－１００凝胶过滤，得凝血酶制品，经ＳＤＳ－ＰＡＧＥ分析，制品呈单一条带，分子量约为３９ｋＤ，比活达１６１０ＮＩＨＵ／ｍｇ蛋白，以活化的粗品活性为１００％，活性回收率为４８％，纯化倍数约为１０。The isolation, purification and characterization of pig thrombin were described.Theprocedure includes isoelectric precipitation of euglobulin at low ionic strength,extractiOn of pro-thrombin from the precipitate,adsorption on magnesium hydroxide and subsequent elution with car-bon dioxide at normal atmosphere,activation of prothrombin,ethanol fractionation,chromatographicpurification successively with DEAE-cellulose,phosphocellulose and Sephadex G-100 columns.Thepurified thrombin showed a single band with molecular weight of 39 kD by SDS-PAGE and twobands with isoelectric point values of 5.0 and 5.4 respectively by PAGIF. The specific activity ofthe purified preparation was 1610 NIH U/mg protein.[

关 键 词：猪血 凝血酶 分离 纯化 分子量 

分 类 号：TQ464.8[化学工程—制药化工]