Enzyme separation by parametric pumping

Affinity chromatography and parametric pumping were combined to reduce trypsin concentration in an aqueous solution. The trypsin inhibitor, chicken ovomucoid, was covalently bonded to Sepharose 4B beads. A solution was cycled over this packing in a column at a low p H in one direction followed by a high p H in the other direction. Trypsin retention was favored at the high p H and elution at the low p H. The decrease in trypsin concentration at one end of the column was fitted to an equation derived from the equilibrium parametric pumping model and was a function of the p H limits imposed on the column. Separation was much less than predicted by equilibrium data, but the equation based on trypsin reduction agreed with the α‐chymotrypsin‐trypsin separation.