The separation process of proteins in a column packed with an ion exchanger PBE94 under the internal pH gradient was simulated experimentally by the elution of model proteins and by the computer calculation.
Under the elution of eluent polybuffer, the linear pH gradient was developed and the elution pH's of myoglobin I and II were observed near the PI's of each proteins. For myoglobin I, the protein fed at different time, was focused into single peak at elution, and the elution times of model proteins appeared near their isoelectric points.
For the smooth and linear pH gradients of the systems having interaction with ion exchangers and mixed buffers, a differential model with the regulation by buffers were proposed incorperating a mixed model buffers. The calculated linear pH gradients showed good agreements with experimental results and sensitive to dimensionless groups and Ke.
In computer simulation of the protein separation and focusing, the coupled differential model with the pH gradient and the protein elution were also proposed. According to our calculation, the protein elution and separations were not only influenced by the Peclet number but also dimensionless number PO but the internal patterns of protein elutions indicated the strong interaction of protein and gel.