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Utility of a high coverage phenyl-bonding and wide-pore superficially porous particle for the analysis of monoclonal antibodies and relatedproducts

Lauber, Matthew
Beck, Alain
Published in Journal of Chromatography. A. 2018, vol. 1549, p. 63-76
Abstract A wide-pore silica-based superficially porous material with a high coverage phenyl bonding was evalu-ated for the analysis of monoclonal antibodies and antibody-drug conjugates. This new material is basedon 2.7 um particles having a shell thickness of 0.40 um and average pore size of approximately 450 Å.Various important features of this reversed phase column technology were explored, including kinetic performance for large biomolecules (i.e. speed of analysis, efficiency and peak capacity), recovery ofproteins, selectivity for resolving modifications, and the possibility to reduce the amount of trifluoroacetic acid in the mobile phase. A systematic comparison was also performed with other existing modernwide-pore phases possessing differences in structure/morphology and chemistry.If all these figures of merit are considered, it is clear that this phenyl bonded wide-pore superficially porous stationary phase is one of the most promising materials to have been developed in recent years.Indeed, it offers kinetic performance comparable to the most efficient wide-pore SPP column on themarket. In terms of protein recovery, this new phase was found to be superior to silica-based and silica-hybrid C4 bonded materials, particularly with separations performed at sub-80◦C temperature. Under such conditions, it in fact shows recoveries that are quite similar to a divinyl benzene (DVB) polymer-based material. More importantly, due to its unique, high coverage phenyl bonding, it offers additionalsteric effects and potentially even t-t interactions that yield advantageous selectivity for mAb sub-unit peaks and ADC species as compared to commonly used C4 or C18 bonded phases. Last but notleast, mobile phases consisting of only 0.02–0.05% trifluoroacetic acid can be successfully used with thiscolumn, without significant loss in recovery and peak capacity.
Keywords Superficially porousWide-porePhenyl bondingColumn efficiencyMonoclonal antibodyAntibody-drug-conjugate
PMID: 29602545
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Research group Sciences analytiques
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BOBALY, Balazs et al. Utility of a high coverage phenyl-bonding and wide-pore superficially porous particle for the analysis of monoclonal antibodies and relatedproducts. In: Journal of Chromatography. A, 2018, vol. 1549, p. 63-76. doi: 10.1016/j.chroma.2018.03.043 https://archive-ouverte.unige.ch/unige:103450

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Deposited on : 2018-04-10

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