Temperature Effects in Hydrophobic Interaction Chromatography of Proteins

Abstract

The thermal behavior of five proteins in hydrophobic interaction chromatography (HIC) were investigated in the temperature range from 0 to 50°C. The conformational change of protein was characterized by the parameter, Z, in the stoichiometric displacement model for retention (SDM‐R). The thermodynamic parameters (ΔH°, ΔS°, ΔC _p ° and ΔG°) of these proteins were determined. It was found that the retention process of protein in HIC is entropy driven, and enthalpy or entropy change of unfolding protein correlated linearly with temperature. The existence of enthalpy and entropy convergence with temperature was also confirmed. The differences of the isoentropic and isoenthalpic temperature for protein unfolding in HIC system from the traditional solution were elucidated.

Keywords:

• Column liquid chromatography
• Hydrophobic interaction chromatography
• Protein unfolding
• Thermodynamic convergence

Acknowledgment

This work was financially supported by the National Natural Science Foundation of China (No. 20575052).