弱阳离子交换色谱中疏水作用对蛋白保留的影响  

Effect of Hydrophobic Interaction on Protein Retention in Weak-cation Exchange Chromatography

作　　者：孙萱[1] 杨云[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,陕西西安710069

出　　处：《分析科学学报》2010年第1期6-10,共5页Journal of Analytical Science

摘　　要：选取了四种常用的弱阳离子交换(WCX)商品柱以研究标准蛋白在其上的色谱保留行为。发现在疏水色谱(HIC)模式下,蛋白在这四种WCX商品柱上也有不同程度的保留特征,且洗脱曲线呈现出保留值随盐浓度变化的"U"型。从分子力学角度定性解释了因疏水相互作用力的存在影响了蛋白在WCX色谱柱上洗脱顺序的改变。运用计量置换理论(SDT)中的两组线性方程进一步证实了WCX和HIC中蛋白与固定相间相互作用力的性质,在HIC中为非选择性作用力,而在离子交换色谱(IEC)中为选择性作用力。这四种色谱柱中的两种事实上可在WCX和HIC两种模式下,对标准蛋白进行分离且有较好的分离效果,有可能作为二维色谱柱来使用。The retention behavior of proteins was investigated by using four commercial columns which have been popularly used in weak-cation exchange （WCX）. It was found that all proteins have the characters of hydrophobic interaction chromatography （HIC） mode on these WCX columns, but to different extents. The obtained elution curves exhibited ＂U＂ shape when the retention time was polted vs. the changes in the salt concentration in the employed mobile phase. This results indicates that the existence of the hydrophobic interactions contributes to the protein retention and may change the eluted order of protein on these WCX columns. This phenomenon was qualitatively explained from the standpoint of molecular mechanism. In addition, two linear equations in the stoichiometric displacement theory （SDT） were employed to test the character of interaction forces between the proteins and the stationary phase. A good linearity indicates that the interaction force is non-selective for HIC mode, while non-linearity expresses that the interaction force is selective force for WCX mode. Actually two of the four columns may be used for protein separation with both WCX and HIC modes, which can be proved from the resolution for protein separation. Each of these two columns may be employed for a two dimensional liquid chromatographic column for protein fast separation.

关 键 词：离子交换色谱 疏水色谱 计量置换 混合保留机理 蛋白分离 

分 类 号：O657.75[理学—分析化学]