RAPID ION-EXCHANGE DISPLACEMENT CHROMATOGRAPHY OF PROTEINS ON PERFUSIVE CHROMATOGRAPHIC SUPPORTS

Displacement chromatography is an extremely powerful separation techni que. However, it has not been widely accepted as a preparative techniq ue due to the creeping flow-rates at which displacement separations ha ve been demonstrated. In this work, the enhanced mass transport of per fusive chromatographic particles enabled displacement separations to b e dramatically scaled-up with respect to flow-rate. At a flaw-rate of 4 ml/min (1440 cm/h) on an analytical-scale column, crude beta-lactogl obulin was displaced in only 90 s to yield ca. 18 mg each of pure beta -lactoglobulin A and B. This represents an order of magnitude decrease in the time required for displacement separations. Thus, the operatio n of perfusive columns in the displacement mode offers an ultra-high t hroughput separation technique with outstanding resolving power even u nder heavily overloaded conditions.