离子交换层析在人血白蛋白生产中的应用研究

目的 研究离子交换层析在人血白蛋白生产中的应用.方法 将采用低温乙醇分离工艺分离的组分Ⅳ上清液超滤脱醇后,采用离子交换层析进行纯化.筛选层析用凝胶并优化离子交换层析的样品pH、电导率和上样量.同时采用高效液相色谱检测人血白蛋白中间品和成品的纯度.结果 选用Capto DEAE凝胶作为层析用介质,最适层析条件为:样品pH值6.9、电导率5.0 mS/cm、上样量≤1.2g蛋白/ml凝胶.高效液相色谱检测显示,优化低温乙醇工艺制备的人血白蛋白中间品纯度可达99.44％～100.00％,而且优化低温乙醇工艺制备的人血白蛋白成品纯度(97.77％)高于传统低温乙醇工艺(93.82％).结论 该离子交换层析工艺可在人血白蛋白生产中应用.

Application study on ion exchange chromatography in human serum albumin manufacture process

Objective To study ion exchange chromatography application in human serum albumin (HSA) manufacture process.Methods The factor Ⅳ supernatant separated by cold ethanol fractionation was dealcoholized by ultrafiltration and purifed by ion exchange chromatography.The column gel was selected,and sample pH,conductivity and sample load for ion exchange chromatography were optimized.The purity of HSA intermediates and final products were detected by high performance liquid chromatography (HPLC).Results Capto DEAE gel was used for ion exchange chromatography,and the optimized chromatographic conditions were sample pH6.9,conductivity 5 mS/cm and sample load ≤1.2 g protein/ml gel.HPLC results showed that the purity of HSA intermediates prepared by optimized cold ethanol fractionation were 99.44％-1.00.00％,and the purity of HSA final products prepared by optimized cold ethanol fractionation (97.77％) was higher than that by traditional cold ethanol fractionation (93.82％).Conclusion Ion exchange chromatography can be applied in HSA manufacture process.