蛋白质在合成阳离子交换剂上的色谱特性研究

用国产材料按间接法合成了螯合型弱阳离子交换剂,详细研究了合成填料的色谱性能,并与商品柱的分离效能进行了比较;在宽温度范围内研究了蛋白质在弱阳离子交换系统中的色谱热力学,测定了蛋白质在色谱过程变性时的热力学参数 (△H0和△S0) 和补偿温度β,提出用标准熵变△S0判断蛋白质的构象变化和用△H0与△S0的补偿关系鉴定蛋白质各变体在色谱系统保留机理的同一性。考察了螯合型弱阳离子交换剂与金属离子的作用,研究了蛋白质在金属螯合色谱中的保留机理。

Studies of Chromatographic Behaviors of Proteins on Synthesized Cation-Exchanger

A chelating weak cation-exchanger has been synthesized by indirect method using domestic materials. The chromatographic performance of the synthesized packing was studied in detail, and compared with separation efficiency of commercial columns (Shim-Pack WCX-1 and Poly CATA). The chromatographic thermodynamics of proteins in weak cation-exchange system was studied over a wide temperature range. The thermodynamic parameters (deltaH0 and deltaS0) and the compensation temperature (beta) at protein denaturation were determined by their chromatographic thermal behaviors. By using the obtained standard entropy change (deltaS0), the conformational change of proteins can be judged. The compensation relationship between deltaH0 and deltaS0 was used to identify the identity of individual protein retention mechanism in chromatographic system. The interaction between chelating weak cation-exchanger and metal ions was examined. The retention mechanism of protein in metal chelating chromatography was explored.