Abstract
A preparative separation method with dynamic equilibrium gradient is proposed. The equilibrium gradient is generated by counteracting chromatographic and electrophoretic forces in the column. The electrophoretic force has a gradient along the column due to the pH gradient induced by applied voltage. Protein continuously introduced in the column is separated and concentrated at an equilibrium point. Experiments were carried out with human hemoglobin, which was purified and concentrated in the column. A stable accumulation zone was formed in the column. A theoretical model was developed to simulate the concentration profile of the protein in the column. An equation to evaluate the position of the accumulation zone was derived. The method was found to be a promising approach to preparative separation.
