title: Combined Yamamoto approach for simultaneous estimation of adsorption isotherm and kinetic parameters in ion-exchange chromatography.
abstract: Application of model-based design is appealing to support the development of protein chromatography in the biopharmaceutical industry. However, the required efforts for parameter estimation are frequently perceived as time-consuming and expensive. In order to speed-up this work, a new parameter estimation approach for modelling ion-exchange chromatography in linear conditions was developed. It aims at reducing the time and protein demand for the model calibration. The method combines the estimation of kinetic and thermodynamic parameters based on the simultaneous variation of the gradient slope and the residence time in a set of five linear gradient elutions. The parameters are estimated from a Yamamoto plot and a gradient-adjusted Van Deemter plot. The combined approach increases the information extracted per experiment compared to the individual methods. As a proof of concept, the combined approach was successfully applied for a monoclonal antibody on a cation-exchanger and for a Fc-fusion protein on an anion-exchange resin. The individual parameter estimations for the mAb confirmed that the new approach maintained the accuracy of the usual Yamamoto and Van Deemter plots. In the second case, offline size-exclusion chromatography was performed in order to estimate the thermodynamic parameters of an impurity (high molecular weight species) simultaneously with the main product. Finally, the parameters obtained from the combined approach were used in a lumped kinetic model to simulate the chromatography runs. The simulated chromatograms obtained for a wide range of gradient lengths and residence times showed only small deviations compared to the experimental data.
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland.
authors: Matthias
authors: M
authors: Rüdt
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland.
authors: Florian
authors: F
authors: Gillet
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland.
authors: Stefanie
authors: S
authors: Heege
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland.
authors: Julian
authors: J
authors: Hitzler
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland.
authors: Bernd
authors: B
authors: Kalbfuss
authors: Novartis Pharma AG, Biologics Process R&D, Klybeckstrasse 141, 4057 Basel, Switzerland. Electronic address: bertrand.guelat@novartis.com.
authors: Bertrand
authors: B
authors: Guélat
mesh_terms: D000327:Adsorption; D000837:Anion Exchange Resins; D000911:Antibodies, Monoclonal / Q000032:analysis; D002411:Cation Exchange Resins; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods*; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007700:Kinetics; D008956:Models, Chemical; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D013816:Thermodynamics
chemical_list: D000837:Anion Exchange Resins; D000911:Antibodies, Monoclonal; D002411:Cation Exchange Resins; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins
keywords: Adsorption equilibrium; Chromatography; Ion-exchange; Mass transfer; Parameter estimation; Protein