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This paper is part of our continued effort to understand the underlying principles of dynamic field gradient focusing. In this investigation, we examined three problems associated with the use of a semi-permeable membrane. First, the influence of steric and ionic exclusion of current carrying ions through the membrane was examined. It was found that resistance to the transport of ions across the membrane resulted in a shallowing of the electric field profile and an increase in the size of the defocusing zone, which is where the slope of the electric field is reversed so that it disperses rather than concentrates solutes. These problems could be reduced by using a membrane with large pores relative to the size of the buffering ions and completely void of fixed charges. Next, a numerical simulation was used to investigate concentration polarization of protein onto the surface of the membrane. Due to the presence of a transverse electric field, species were pulled toward the membrane. If the membrane is restrictive to those species, a concentrated, polarized layer will form on the surface. The simulation showed that by decreasing the channel to a depth of 20 microm, the concentrated region next to the membrane could be reduced. Finally, it was found that changes in column volume due to loss of membrane structural integrity could be mitigated by including a porous ceramic support. The variation in peak elution times was decreased from greater than 20% to less than 3%.
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