High-performance liquid chromatography of amino acids, peptides and proteins: CXII. Analysis of operating parameters affecting the breakthrough curves in fixed-bed chromatography of proteins using several mathematical models

Abstract

Performance of a packed bed, in terms of adsorbent utilization and kinetic characteristics, was studied for the adsorption of HSA to the ion-exchange resins, DEAE Trisacryl M and DEAE Sepharose FF. The influence of column length, temperature and feed concentration on the shape of the breakthrough curve has been investigated and the resulting operating capacities, q * measured by graphical integration. Mathematical models prevalent in the literature have been used to extract out the kinetic parameters, the effective diffusivity, D p, and the interaction rate constant, k 1, from the experimental breakthrough curves, and these parameters have been used to evaluate the performance of the columns and to validate the models selected. The effect of protein size on the adsorption behaviour was also investigated using the proteins HSA, transferrin and ferritin, determining q *, k 1, and D p from the breakthrough curves.