摘要
制备了弱阴离子交换整体柱,并建立了白介素 18在该柱上分离纯化的方法。以色谱柱管为模具,通过原位聚合法制备了一种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)为交联剂的整体色谱柱,并用二乙胺将其修饰为弱阴离子交换柱。考察了交联剂及致孔剂用量对色谱柱压力的影响,发现:随着交联剂用量的减小,致孔剂用量的增大,柱压下降;当交联剂和致孔剂用量达到一定水平后,柱压变化不再明显,但柱体机械强度降低。用该柱对白介素 18进行了分离,同时考察了不同固定相、缓冲液体系、pH值、盐的类型以及有机添加剂对白介素 18分离效果的影响。方法简单、灵敏、快速,重现性好。
A weak anion exchange column was prepared and a separation method of interleukin-18 (IL-18) was developed on the column. The chromatographic column with glycidyl methacrylate as functional monomer and ethylene dimethacrylate as cross-linker was prepared by a free radical polymerization within a 64 mm×4.6 mm i.d. column. The epoxide groups of the column were modified by a reaction with diethylamine that affords ionizable functionalities (required) for ion exchange chromatography. The influences of the contents of the cross-linker and porogen on column pressure were investigated. The factors influencing the separation, such as stationary phases, buffers, pH values, salts, and contents of organic solvent, were studied. The optimal conditions for separating IL-18 from its related proteins were obtained. The (proposed) method is simple, sensitive, rapid and reproducible.
出处
《色谱》
CAS
CSCD
北大核心
2005年第2期168-171,共4页
Chinese Journal of Chromatography
基金
国家自然科学基金项目 (No. 20375010 )
河北省自然科学基金项目 (No. 202096 )
科学技术部基金项目 (No.2002CCA3100
KJCX2 H4)
中国科学院百人计划项目.
关键词
弱阴离子交换色谱
整体柱
白介素-18
weak anion exchange chromatography
monolithic column
interleukin-18
