Chromatographic Behaviors of Proteins on Cation‐Exchange Column

AbstractA weak cation‐exchanger (XIDACE‐WCX) has been synthesized by the indirect method. The chromatographic characteristics of the synthesized packing was studied in detail. The standard protein mixture and lysozyme from egg white were separated with the prepared chromatographic column. The chromatographic thermodynamics of proteins was studied in a wide temperature range. Thermodynamic parameters standard enthalpy change (ΔH0)and standard entropy change (ΔS0) and compensation temperature (β�) at protein denaturation were determined in the chromatographic system. By using obtained ΔS0, the conformational change of proteins was judged in the chromatographic process. The linear relationship between ΔH0 and ΔS0 can be used to identify the identity of the protein retention mechanism in the weak cation‐exchange chromatography. The interaction between weak cation‐exchanger and metal ions was investigated. Several metal chelate columns were prepared. The effects of introducing metal ion into the naked column on protein retention and the retention mechanism of proteins in the metal chalet affinity chromatography were discussed.