反离子对牛血清白蛋白在荷电葡聚糖接枝介质中色谱行为的影响  

Effect of counterions on protein adsorption and elution of DEAE-dextran modified Sepharose FF

作　　者：韩德涛 白姝[1] 龚玲莉 余林玲 孙彦[1] 

机构地区：[1]天津大学化工学院生物化工系系统生物工程教育部重点实验室,天津300072

出　　处：《化工进展》2018年第1期260-268,共9页Chemical Industry and Engineering Progress

基　　金：国家自然科学基金项目(21406160;21236005;21621004)

摘　　要：从前期开发的具有极高的吸附容量及传质速率的二乙氨乙基葡聚糖接枝离子交换介质中选取FF-D50-Dex D100和FF-Dex D100为典型代表,利用Cl~–、SCN~–、SO_4^(2–)、HPO_4^(2–)为模型反离子,以牛血清白蛋白(BSA)为模型蛋白,以商品化介质(Q Sepharose FF、Q Sepharose XL、DEAE Sepharose FF)为对照,在离子强度为0.06mol/L下,系统研究反离子对二乙氨乙基葡聚糖接枝介质的蛋白质吸附与洗脱行为的影响。结果表明,二乙氨乙基葡聚糖接枝介质对不同反离子的偏好性存在差异,且该偏好性差异与基团所处位置(接枝链配基或表面配基)无关。同时,介质偏好性弱的反离子会通过促进二乙氨乙基葡聚糖接枝介质的"链传递"效应加快蛋白质的传质速率,从而提高动态吸附容量。因此,在使用二乙氨乙基葡聚糖接枝介质进行蛋白质色谱柱分离过程中,可在吸附操作中使用HPO42–,在洗脱操作中使用SCN–来优化分离效果。In the previous studies,a series of novel anion-exchangers,DEAE-dextran grafted Sepharose FF resins,were obtained,which exhibited high levels of both protein capacity and uptake rate.In this work,two typical DEAE-dextran grafted resins,FF-D50-Dex D100（mixed-ligand resin） and FF-Dex D100（grafting-ligand resin）,were selected to investigate the effects of counterions on protein adsorption equilibria,uptake rate,dynamic binding and linear gradient eltion,with sodium salts of SCN–,Cl–,HPO42– and SO42–,and compared with the commercial resins,Q Sepharose FF,Q Sepharose XL and DEAE Sepharose FF,using bovine serum albumin（BSA） as the model protein.It was found that the four counterions showed different prefernces on the two DEAE-dextran modified resins,but the couterion preference orders of two DEAE-dextran modified resins were the same,which indicated that the ligand distribution（surface-ligand or grafting-ligand） did not affect the couterion preference.Furthermore,the counterions with weaker prefernces could accelerate the masstransfer of protein by promoting the ＂chain delivery＂ effect,and finally enchance the dynamic binding capacity of DEAE-dextran modified resins.Therefore,HPO42– and SCN– are suitable counterions for column binding and eltuion operation with DEAE-dextran grafted Sepharose FF resins,respecitively.The experimental result is expected to help optimizing anion exchange chromatograhpy process with DEAE-dextran modified resins.

关 键 词：离子交换 二乙氨乙基葡聚糖 反离子 吸附 动力学 线性梯度洗脱 

分 类 号：TQ936.2[化学工程] TQ317.3