Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 1: alternative mobile phases and fine tuning of the separation

Publication date: Available online 19 February 2019Source: Journal of Pharmaceutical and Biomedical AnalysisAuthor(s): Evelin Farsang, Amarande Murisier, Krisztián Horváth, Alain Beck, Róbert Kormány, Davy Guillarme, Szabolcs FeketeAbstractCation exchange chromatography (CEX) of therapeutic monoclonal antibodies is generally performed with either salt gradient (MES buffer + NaCl) or using commercial pH gradient buffer. The goal of this study was to find out some alternative buffer systems for CEX separation of mAbs, which may offer alternative selectivity, while maintaining similar peak shapes. Among the new buffers that were tested, (N-morpholino)ethanesulfonic acid (MES) / 1,3-diamino-2-propanol (DAP), and citric acid / 2-(cyclohexylamino)ethanesulfonic acid (CHES) systems were particularly promising, especially when combining them with a moderate salt gradient of NaCl. This two buffer system provides an equivalent or slightly better separation than the standard, mobile phases for therapeutic mAbs.It was also demonstrated that working with salt-mediated pH gradients, allows to extend the possibilities in method development, since the concentration of salt in the mobile phase has a significant impact on selectivity. Using HPLC modeling software (Drylab), it was possible to successfully develop CEX methods for authentic mAb samples within only 6 hours, by optimizing the gradient steepness and salt concentration in the B eluent.
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research

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SURESH K. MUKHERJI, MD, MBA, FACR
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Source: Neuroimaging Clinics - Category: Radiology Authors: Source Type: research
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