Study of the retention mechanism of proteins in hydrophobic interaction chromatography

摘要: A stoichiometric displacement retention model for proteins based on hydrophobic interaction chromatography (HIC) is presented. Several methods were used to demonstrate that water the displacing agent in this process. Salt not only affects molar concentration of water, but also changes confomation proteins, forces and number molecules a series hydrated protein molecules. An equation relates capacity factor k′, factor, Z (the required displace from ligands) was derived. The intercept equation, log I, contains constants relate affinity ligands. There good linear relationship between k′ [H2O] under different chromatographic conditions. Although I varied with pH, salt temperature, plot vs. always an excellent straight line, slope j ≈ 1.74, logarithm pure water. Hydrophobic interations dominate HIC at high concentrations.