以强阴离子交换剂FG TMAE为介质的离子交换层析制备人凝血因子Ⅷ的初探

摘要

Objective To explore preparation of human coagulation factor Ⅷ (FⅧ)using ion exchange chromatography (ICE) with a strong anion exchanger FG TMAE as separating medium.Methods The initial conditions of ICE were identified by linear gradient elution using eluent containing different concentrations of NaCl.Ionic strength and pH value of the wash solution and eluentand and temperature of the sample were optimized to finally identify the optimum conditions of ICE with FG TMAE.Results When ICE was carried out using wash solution containing 220 mmol/L NaCl and eluent containing 450 mmol/L NaCl at pH7.0 in room temperature (20-25 ℃),recoveries (77.53％) and specific activity (24.67 IU/mg) of FⅧ were both high.Conclusion FⅧ preparation using ICE with FG TMAE is feasible.%目的 初步探索以强阴离子交换剂FG TMAE为介质进行离子交换层析来制备人凝血因子Ⅷ.方法 以含不同氯化钠浓度的洗脱液进行线性梯度来初步确定离子交换层析的条件,并优化离子交换层析的洗涤液和洗脱液的离子强度和pH值以及样品的温度,最终确定以强阴离子交换剂FGTMAE为介质进行离子交换层析的最适条件.结果 在室温(20～25℃)条件下,以pH 7.0的含220 mmol/L氯化钠的洗涤液和pH 7.0的含450 mmol/L氯化钠的洗脱液进行离子交换层析,可使人凝血因子Ⅷ的回收率(77.53％)和比活性(24.67 IU/mg)均较高.结论 以强阴离子交换剂FG TMAE为介质进行离子交换层析制备人凝血因子Ⅷ是可行的.