VERY HIGH-SPEED SEPARATION OF PROTEINS WITH A 20-MU-M REVERSED-PHASE SORBENT

A five-protein mixture was successfully separated in less than 15 s by reversed-phase gradient elution on a 20-mu-m polymeric flow-through type chromatographic packing material (POROS R/M). The same mixture could also be separated with far better resolution in less than 60 s on the same column by simply extending the gradient volume. Pressure drop across the column was less than 130 bar, despite a superficial linear velocity of nearly 9000 cm/h. Frontal chromatography at different flow-rates was used to demonstrate that the ability to separate at high speed was due to greatly enhanced mass transport within the particles due to the flow-through (perfusion chromatography) effect.