CAJ | 학술논문

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.
선취료사충상용적약양리자교환(WCX)상품주이연구표준단백재기상적색보보류행위.발현재소수색보(HIC)모식하,단백재저사충WCX상품주상야유불동정도적보류특정,차세탈곡선정현출보류치수염농도변화적"U"형.종분자역학각도정성해석료인소수상호작용력적존재영향료단백재WCX색보주상세탈순서적개변.운용계량치환이론(SDT)중적량조선성방정진일보증실료WCX화HIC중단백여고정상간상호작용력적성질,재HIC중위비선택성작용력,이재리자교환색보(IEC)중위선택성작용력.저사충색보주중적량충사실상가재WCX화HIC량충모식하,대표준단백진행분리차유교호적분리효과,유가능작위이유색보주래사용.
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.