title: Characterization of cation exchanger stationary phases applied for the separations of therapeutic monoclonal antibodies.
abstract: Cation exchange chromatography (CEX) is a well established strategy for the characterization of monoclonal antibodies (mAbs). The optimization of mobile phase conditions is well described in the literature, but there is a lack of information about CEX stationary phases for the analysis of therapeutic proteins. The aim of this study was to compare five state-of-the-art CEX stationary phases based on the retention, selectivity and resolving power achieved in pH- and salt-gradient modes, with various therapeutic mAbs and their variants. The Sepax Antibodix WCX-NP3, Thermo MAbPac SCX-10 RS, YMC BioPro SP-F, Waters Protein-Pak Hi Res SP and Agilent Bio mAb NP1.7 SS were considered in this study. In terms of retention, the YMC Bio Pro SP-F material was the less retentive one, while the Agilent Bio mAb NP1.7 SS provides the highest retention. Regarding the selectivity achieved between the main mAbs isoforms and their variants, the Thermo MabPac SCX column generally gave the highest selectivity. Finally, it was hard to rank columns in term of kinetic performance since their performance is strongly solute (mAb) and elution mode (pH or salt gradient) dependent. However, the highest resolution--in most cases--was observed on the strong cation exchanger YMC Bio Pro SP-F material.
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Boulevard d'Yvoy 20, 1211 Geneva 4, Switzerland. Electronic address: szabolcs.fekete@unige.ch.
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Fekete
authors.affiliation: Center of Immunology Pierre Fabre, 5 Avenue Napoléon III, BP 60497, 74160 Saint-Julien-en-Genevois, France(1).
authors.forename: Alain
authors.initials: A
authors.lastname: Beck
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Boulevard d'Yvoy 20, 1211 Geneva 4, Switzerland.
authors.forename: Davy
authors.initials: D
authors.lastname: Guillarme
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002412:Cations / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods; D006863:Hydrogen-Ion Concentration; D012965:Sodium Chloride / Q000737:chemistry; D012996:Solutions / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D002412:Cations; D012996:Solutions; D012965:Sodium Chloride
keywords: Cetuximab; Column coupling; Ion exchange; Monoclonal antibody; Stationary phase