关于混合模式色谱固定相对蛋白保留特征的研究  

Studies on the Characterization of Stationary Phase for Protein Retention on Mixed-mode Chromatography

作　　者：加晓丹[1] 柯从玉[2] 孙萱[1] 耿信笃[1] 

机构地区：[1]西北大学现代分离科学研究所,现代分离科学省级重点实验室,教育部合成与天然功能分子化学重点实验室,西安710069 [2]西安石油大学化学化工学院,西安710065

出　　处：《化学学报》2012年第15期1631-1636,共6页Acta Chimica Sinica

基　　金：陕西省重点实验室基金(No.11JS099)资助~~

摘　　要：为使混合机理色谱(MMC)得到广泛地应用,合成、表征和评价MMC固定相就成了首先要解决的问题.依据离子交换色谱柱也具有疏水色谱(HIC)保留机理的特征,选了4种弱阳离子交换(WCX)柱和一根二维[2D(WCX,HIC)]色谱柱,研究了标准蛋白在这两类色谱柱上的保留行为.这四种WCX色谱柱中的两种能在WCX和HIC两种分离模式下分离蛋白,虽不如2D色谱柱效果好,但有可能当成"准2D柱"来使用.发现蛋白在这四种WCX柱上所显示的HIC分离特征各不相同,且保留值随盐浓度变化呈现出的"U型"曲线也有大的差异.实验结果显示,"U型"曲线的宽度和临界点分别与色谱动力学和热力学因素相关.还对这两类色谱固定相的峰容量表征方法和命名提出了建议和说明.Mixed-mode chromatography（MMC） is a type of chromatographic method in which multiple-mode take place between the stationary phase and solute in the feed.MMC has been found to have many advantages and thus became a research hot point.However,before going wide applications,the synthesis of new MMC column and/or the selection of some commercial columns having the character of MMC need to be established.Based on the fact that some commercial ion-exchange columns perform the separation behavior of hydrophobic interaction chromatography（HIC）,one of four commercial weak-cation exchange（WCX） column,a two-dimensional mode（WCX,HIC） column called as 2D column,and standard proteins were selected to investigate and compare their retention behaviors.The two type columns have some common characters： proteins can be separated by either WCX,or HIC mode,but have some quite different column efficiencies,and the profiles of ＂U-shape＂ elution curve.The width of the ＂U-shape＂ curve was experimentally found to relate to the chromatographically dynamic factor,i.e.,the broader the ＂U-shape＂ curve,the narrower the chromatographic peak obtains,while the magnitude of the critical point of the ＂U-shape＂ curve relates to the protein retention,a larger value of a critical point corresponds to a stronger retention.An intensive comparison between the two types of column efficiency was carried out by the peak capacity of proteins.The experimental results showed that the 2D column has much more number of peak capacity than the commercial WCX column,either for one-dimensional,or two-dimensional separation.However,for the separation of a group proteins,the elution order is dominated by the comprehensive consideration of the peak capacity,the dynamic,and thermodynamic factors.Finally the nomenclature of the various stationary phases of MMC and/or 2D column employed in two-dimensional liquid chromatography were suggested.Although,in terms of column efficiency,none of the four commercial WCX column could be compared to

关 键 词：二维色谱固定相 混合模式色谱 蛋白分离 离子交换色谱 疏水色谱 计量置换理论 

分 类 号：O657.7[理学—分析化学]