High-performance affinity chromatography of human thrombin on modified polystyrene resins.

Cross-linked polystyrenes modified with L-arginyl methyl ester mimic the binding site of antithrombin III and thrombin substrates. They can be used as stationary phases in high-performance affinity chromatography of thrombin. Under isocratic conditions, thrombin is strongly adsorbed on the resins when the sodium chloride concentration is lower than 0.5 M. The bound enzyme can be selectively desorbed when the salt concentration is raised to about 1.2 M. With a linear salt gradient, the specific elution of thrombin can be effected with a high recovery of its enzymatic activity. The decomposition products of thrombin, when treated with sodium dodecyl sulphate, are not retained by the stationary phase. The effects of the flow-rate and salt gradient slope on the adsorption and desorption of alpha-thrombin demonstrate the importance of kinetic parameters.