ISOELECTRIC-FOCUSING FIELD-FLOW FRACTIONATION .4. INVESTIGATIONS ON PROTEIN SEPARATIONS IN THE TRAPEZOIDAL CROSS-SECTION CHANNEL

The first separation of three proteins (horse spleen ferritin, equine myoglobin and horse heart cytochrome c) by isoelectric focusing field-flow fractionation in a trapezoidal cross-section channel of 0.875 ml volume and 25 cm length is reported. Separation and elution are shown to proceed within about 1 h at a power application of about 1 W. The separation of the three proteins is demonstrated to be dependent on applied electric power, carrier ampholyte concentration and the concentrations of anolyte and catholyte. It follows from these data that the resolution is improved with increasing carrier ampholyte concentration and/or decreasing concentrations of the electrode solutions. The experimentally observed effects are in agreement with predictions made by computer simulation.