title: Prototype sphere-on-sphere silica particles for the separation of large biomolecules.
abstract: The goal of this study was to evaluate the possibilities offered by a prototype HPLC column packed with ∼2.5μm narrow size distribution sphere-on-sphere (SOS) silica particles bonded with C4 alkyl chains, for the analytical characterization of large biomolecules. The kinetic performance of this material was evaluated in both isocratic and gradient modes using various model analytes. The data were compared to those obtained on other widepore state-of-the-art fully core-shell and fully porous materials commonly employed to separate proteins moreover to a reference 5μm wide pore material that is still often used in QC labs. In isocratic mode, minimum reduced plate height values of hmin=2.6, 3.3 and 3.3 were observed on butylparaben, decapeptide and glucagon, respectively. In gradient elution mode, the SOS column performs very high efficiency when working with fast gradients. This prototype column was also comparable (and sometimes superior) to other widepore stationary phases, whatever the gradient time and flow rate, when analyzing the largest model protein, namely BSA. These benefits may be attributed to the SOS particle morphology, minimizing the intra-particle mass transfer resistance. Finally, the SOS column was also applied for the analytical characterization of commercial monoclonal antibody (mAb) and antibody-drug conjugate (ADC) samples. With these classes of proteins, the performance of SOS column was similar to the best widepore stationary phases available on the market.
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Bd d'Yvoy 20, 1211 Geneva 4, Switzerland. Electronic address: szabolcs.fekete@unige.ch.
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Fekete
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Bd d'Yvoy 20, 1211 Geneva 4, Switzerland. Electronic address: marta.rodriguezaller@unige.ch.
authors.forename: Marta
authors.initials: M
authors.lastname: Rodriguez-Aller
authors.affiliation: Department of Pharmacy, University of Bologna, Bologna, Via S. Donato 15, 40127, Italy. Electronic address: alessandra.cusumano@studio.unibo.it.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Cusumano
authors.affiliation: Department of Chemistry, University of Liverpool, Crown Street, Liverpool L69 7ZD, United Kingdom. Electronic address: Richard.Hayes@liverpool.ac.uk.
authors.forename: Richard
authors.initials: R
authors.lastname: Hayes
authors.affiliation: Department of Chemistry, University of Liverpool, Crown Street, Liverpool L69 7ZD, United Kingdom.
authors.forename: Haifei
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Thermo Fisher Scientific, Runcorn, United Kingdom. Electronic address: tony.edge@thermofisher.com.
authors.forename: Tony
authors.initials: T
authors.lastname: Edge
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Bd d'Yvoy 20, 1211 Geneva 4, Switzerland. Electronic address: jean-luc.veuthey@unige.ch.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Veuthey
authors.affiliation: School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Bd d'Yvoy 20, 1211 Geneva 4, Switzerland. Electronic address: davy.guillarme@unige.ch.
authors.forename: Davy
authors.initials: D
authors.lastname: Guillarme
mesh_terms: D000911:Antibodies, Monoclonal; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D007700:Kinetics; D010316:Particle Size; D016062:Porosity; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D012822:Silicon Dioxide
keywords: Brentuximab vedotin; Column efficiency; Fractal column; Monoclonal antibody; Rituximab; Sphere-on-sphere particles