title: Electrostatic model for protein adsorption in ion-exchange chromatography and application to monoclonal antibodies, lysozyme and chymotrypsinogen A.
abstract: A model for the adsorption equilibrium of proteins in ion-exchange chromatography explicitly accounting for the effect of pH and salt concentration in the limit of highly diluted systems was developed. It is based on the use of DLVO theory to estimate the electrostatic interactions between the charged surface of the ion-exchanger and the proteins. The corresponding charge distributions were evaluated as a function of pH and salt concentration using a molecular approach. The model was verified for the adsorption equilibrium of lysozyme, chymotrypsinogen A and four industrial monoclonal antibodies on two strong cation-exchangers. The adsorption equilibrium constants of these proteins were determined experimentally at various pH values and salt concentrations and the model was fitted with a good agreement using three adjustable parameters for each protein in the whole range of experimental conditions. Despite the simplifications of the model regarding the geometry of the protein-ion-exchanger system, the physical meaning of the parameters was retained.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093 Zurich, Switzerland.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Guélat
authors.forename: Guido
authors.initials: G
authors.lastname: Ströhlein
authors.forename: Marco
authors.initials: M
authors.lastname: Lattuada
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
mesh_terms: D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange / Q000379:methods*; D002919:Chymotrypsinogen / Q000737:chemistry* / Q000302:isolation & purification; D007700:Kinetics; D008956:Models, Chemical; D009113:Muramidase / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D002919:Chymotrypsinogen; D009113:Muramidase