| As a nonlinear chromatographic technique, displacement chromatography has attractive more attention since its introduction in 1943. One of important areas in displacement chromatography is to develop potential displacers. In this article, a novel strategy in displacer design has been proposed based on amino acid-based dendrimers and a library of dendrimeric displacers using amino acid as extending unit and lysine as a branching unit was constructed. The virtual screening of this library of displacer was established on the hydrophobic index (log P). The results in virtual screening indicated that the introduction of phenyl group in displacer would increase the log P significantly. Therefore, three displacers, phenylalanine-based, leucine-based and glycine-based displacers, were synthesized based on the solid-phase peptide synthesis. The MS results exhibited the target products have high purity.Secondly, the dynamic affinities of cytochrome C (Cyt-C),α-chymotrypsinogen A (Chy-A) and displacers have been examined using steric-mass action (SMA) model. The linear SMA parameters of protein have been determined by elution experiments while those of displacers linear gradient elution experiments. Then, nonlinear SMA parameter of proteins was determined by fitting adsorption equilibrium data with SMA model. Based on the SMA model, the plots of dynamic affinity and affinity ranking were given to decide the operation condition and predict the isotachic displacement train of proteins.Finally, multi-component static displacement experiment was used to evaluate the displacement ability of dendrimeric displacers. The results exhibited all of dendrimeric displacers have higher displacement efficiencies in particular for phenylalanine-based displacer. The enhancement of displacement ability is directly due to the introduction of phenyl group and other hydrophobic groups. The current research provided a fundamental guide in the design of potential displacer...
|
PDF Full Text Request