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Poly(ethylene glycol)-based monolithic capillary columns for hydrophobic interaction chromatography of immunoglobulin G subclasses and variants
journal contribution
posted on 2023-05-17, 20:00 authored by Desire, CT, Arrua, RD, Mohammad TalebiMohammad Talebi, Lacher, NA, Emily HilderEmily HilderPolymermonoliths were prepared in 150 um id capillaries by thermally initiated polymerization of PEG diacrylate for rapid hydrophobic interaction chromatography of immunoglobulin G (IgG) subclasses and related variants. Using only one monomer in the polymerization mixture allowed ease of optimization and synthesis of the monolith. The performance of the monolith was demonstrated by baseline resolution of IgG subclasses and variants, including mixtures of the k variants of IgG1, IgG2, and IgG3 as well as the k and ? variants associated with IgG1 and IgG2. The effect of eluent concentration and pH on the separation efficiency of studied proteins was also explored, allowing almost baseline resolution to be achieved for mixtures of the k variants of IgG1, IgG2, IgG3, and IgG4 but also for the k and ? variants of IgG1 and IgG2. The results showed significant improvement in the separations in terms of the tradeoff between analysis time and resolution, while maintaining a simple methodology, in comparison to previous reports. The synthesized monolith was also used for the separation of isoforms of a therapeutic monoclonal antibody.
History
Publication title
Journal of Separation ScienceVolume
36Issue
17Pagination
2782-2792ISSN
1615-9306Department/School
School of Natural SciencesPublisher
Wiley - V C H Verlag GmbH & Co. KGaAPlace of publication
GermanyRights statement
Copyright 2013 WILEY-VCH Verlag GmbH & Co. KGaA, WeinheimRepository Status
- Restricted