羟基磷灰石电色谱中电渗流动与分离过程特性  

ELECTROOSMOTIC FLOW AND PROCESS CHARACTERISTICS OF HYDROXYLAPATITE ELECTROCHROMATOGRAPHY

作　　者：殷钢[1] 李琛[1] 詹劲[1] 袁乃驹[1] 刘铮[1] 

机构地区：[1]清华大学化学工程系,北京100084

出　　处：《化工学报》2001年第8期666-672,共7页CIESC Journal

基　　金：国家自然科学基金 (No .2 96760 2 2 );北京市自然科学基金 (No .2 992 0 16);清华大学曹光彪高科技基金资助项目&&

摘　　要：在羟基磷灰石色谱上施加电场 ,发展了一种新的电色谱分离技术 .对蛋白质电泳和溶液电渗对分离特性的影响进行了实验研究 ,测量了羟基磷灰石电色谱装置中膜、膜与填充介质在不同浓度磷酸缓冲溶液和在不同电流下产生的电渗流 ,分别计算出由膜和羟基磷灰石电渗而产生的推动力 .实验结果表明电渗流量随缓冲溶液离子强度的降低和电流的增加而增加 .在施加电场的情况下 ,羟基磷灰石对牛血清白蛋白的吸附速度加快 ,动态吸附量增加 .分别考察了不同洗脱条件下电场对洗脱效果的影响 .在高浓度的洗脱溶液下 ,电渗的作用很小 ,电场的作用主要是电迁移的作用 ,可以通过选择适宜的电场方向来提高分离精度 ;在低浓度的洗脱溶液下 ,电渗的作用有效地促进了洗脱液与介质颗粒的传质 ,提高了洗脱效果 .在羟基磷灰石色谱上施加电场提高了该色谱分离过程的速率和效率 .A novel kind of preparative electrochromatography, namely hydroxylapatite electrochromatography (HEC), is proposed in which an electric potential is applied to the compartment packed with hydroxylapatite. The effects of electric migration, including the electrophoresis of protein and the electroosmosis of solvent, on the separation performance of HEC are experimentally investigated. Electroosmotic fluxes of membrane, membrane and packed medium are determined and the driving forces by electroosmotic flux of membrane and packed medium are interpreted respectively. An increase in electroosmotic flux is obtained in response to the increase in electric field strength and the decrease in ionic concentration of the buffer. The breakthrough curves obtained in the presence of an electric field show an enhanced adsorption of BSA on hydroxylapatite in terms of available bed volume for adsorption and dynamic adsorption capacity. When the elution of BSA from hydroxylapatite is conducted with the buffer of high salt concentration, the enhancement of mass transfer is mainly contributed by electrophoresis. In this case the suitable selection of polarity of electric field is essential to resolution. When elution is done with the buffer of low salt concentration, the contribution of electroosmotic flux to mass transfer becomes significant. The above results show that the application of electric field to hydroxylapatite chromatography, as shown by HEC, greatly improves its separation throughput and efficiency.

关 键 词：羟基磷灰石 电色谱 电渗 电泳 牛血清白蛋白 吸附 蛋白质 分离 洗脱 

分 类 号：Q51[生物学—生物化学] O658[理学—分析化学]