猪凝血因子Ⅹ及凝血因子Ⅹa的制备工艺研究

目的从猪血中制备猪凝血因子Ⅹ(FⅩ)及凝血因子Ⅹa(FⅩa)。方法采用柠檬酸钡吸附、硫酸铵分级沉淀、DEAE 纤维素离子交换色谱等方法制备FⅩ,并优化FⅩ的激活条件,从而制备FⅩa。通过SDS PAGE进行FⅩ和FⅩa的检测。结果建立了FⅩ和FⅩa的制备工艺,收率分别为每1ml血浆34,12 μg;采用胰蛋白酶激活FⅩ,激活条件为:0 .0 2 %酶浓度、pH 8.0、37℃下作用70min。结论此工艺操作简单,收率高,适合于规模化生产

Preparation of porcine factor Ⅹ and factor Ⅹ a

PurposeTo extract porcine factor Ⅹ(FⅩ)and factor Ⅹa(FⅩa)from porcine plasma.MethodsUsing techniques including barium citrate absorption,ammonium sulfate fractionation and DEAE-cellulose chromatography to extract FⅩ,and optimizing the condition of activating FⅩ to FⅩa. FⅩ and FⅩa were tested by SDS-PAGE.ResultsThe preparation method of FⅩ and FⅩa was set up with yield of 34,12 μg per milliliter of starting plasma respectively. FⅩ was activated by trypsin and the optimal activating condition was: adding 0.02% trypsin and incubating for 70 min at 37℃(pH 8.0).ConclusionThis preparative procedure was simple and suitable for mass production with high yield.