弱阳离子交换毛细管整体柱的制备与蛋白质分离  

Preparation of Weak Cation Exchange Monolithic Capillary Column and Its Application for Protein Separation

作　　者：丁明玉[1] 郑睿[1] 彭虹[1] 

机构地区：[1]清华大学化学系,生命有机磷化学及化学生物学教育部重点实验室,北京100084

出　　处：《分析化学》2009年第3期395-398,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20775040)资助项目

摘　　要：以甲基丙烯酸缩水甘油酯(GMA)为单体,乙二醇二甲基丙烯酸酯(EDMA)为交联剂,正丙醇、1,4-丁二醇和水为致孔剂,偶氮二异丁腈(AIBN)为引发剂,在0.32mm内径弹性石英毛细管内原位制备了聚合物整体固定相基质。优化得到的最佳反应物组成为V(GMA):V(EDMA):V(正丙醇):V(1,4-丁二醇):V(H2O)=0.32:0.08:0.35:0.2:0.05,在60℃反应24h,得到了孔径分布在100～300nm的大孔型整体固定相。用亚胺基乙二酸(IDA)对整体柱进行表面改性,制备了羧酸基弱酸型阳离子交换整体固定相。优化得到的最佳改性条件是:反应时间24h、温度75℃和pH12.0。本研究制备的一根仅5cm长的弱阳离子交换毛细管整体柱可以在13min内分离粗卵蛋白、胰蛋白酶和溶菌酶3种模型蛋白质。A polymer-based capillary monolith was prepared by in situ copolymerization using glycidyl methacrylate（GMA） as monomer, ethylenedimethacrylate （EDMA） as cross link agent and 1-propanol, 1,4- butanediol and water as porogen solvent. The optimal ratio of reactants in volume is GMA： EDMA： 1-propanol： 1,4-butanediol： water = 0. 32：0. 08：0. 35：0. 2： 0.05. A macroporous monolith with 100 - 300 nm pore size was obtained reacting for 24 h at 60 ~C. The monolith was modified with iminodiaeetic acid （IDA） and a weak cation-exchange monolithic capillary columns were obtained. The optimal modified conditions were reaction time 24 h, reaction temperature 75 ℃ and reaction pH 12.0. The three model proteins（albumin egg, trypsin and lysozyme） could be separated on this weak cation-exchange monolithic capillary column with a length of only 50 mm within 13 min.

关 键 词：毛细管整体柱 甲基丙烯酸酯 离子交换色谱 蛋白质 

分 类 号：Q51[生物学—生物化学]