Modeling of salt and pH gradient elution in ion-exchange chromatography

摘要: The separation of proteins by internally and externally generated pH gradients in chromatofocusing on ion-exchange columns is a well-established analytical method with large number applications. In this work, stoichiometric displacement model was used to describe the retention behavior lysozyme SP Sepharose FF monoclonal antibody Fractogel SO3 (S) linear salt gradient elution. dependence binding charge B elution introduced using protein net model, while equilibrium constant based thermodynamic approach. parameter dependences are calculated from elutions at different values as well fixed concentrations. application for well-characterized resulted almost identical parameters either or data. For antibody, only approach feasible because limited range useful an acid variant major monomeric form discussed.