Pulse response in an immobilized‐enzyme column: Theoretical method for predicting elution curves

A theoretical method for predicting the elution profile of a pulse response in an immobilized‐enzyme column is proposed. The method is based on a mass balance model, which is extensively used in gel chromatography. To test the method, a pulse of sucrose solution was applied to a column of spherical acrylamide gel in which was entrapped invertase from yeast, and it was eluted with 0.05M acetate buffer at pH 5.0. The elution curves of the substrate and the product were in fairly good agreement with the theoretically calculated ones. The method was extended to the system of reversible as well consecutive reactions.