Optimization of protein separation by continuous-flow electrophoresis: influence of the operating conditions and the chamber thickness.

ABSTRACT

The separation of protein by continuous-flow electrophoresis is disturbed by a number of effects which cause spreading of the protein streams with a loss of resolution. Two numerical models have been used to describe the coupling between the various spreading phenomena. A simple model takes into account molecular diffusion, electroosmosis and residence time gradients; the main model differs from the simple one by taking account of electrohydrodynamics. The influence of the separation chamber thickness, the radius of the sample filament, the field strength, and the residence time is explored. The simulations show that each dispersive effect has its own range of predominance, so that an optimum is found for the thickness of the separation chamber.