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Comparison of various silica-based monoliths for the analysis of large biomolecules

Published in Journal of separation science. 2013, vol. 36, no. 14, p. 2231-2243
Abstract In the present study, three types of silica-based monoliths, i.e. the first and second generations of commercial silica monolithic columns and a wide-pore prototype monolith were compared for the analysis of large biomolecules. These molecules possess molecular weights between 1 and 66 kDa. The gradient kinetic performance of the first-generation monolith was lower than that of the second generation, for large biomolecules (>14 kDa)but very close with smaller ones (1.3–5.8 kDa). In contrast, the wide-pore prototype column was particularly attractive with proteins larger than 19 kDa (higher peak capacity). Among these three columns, the selectivity and retention remained quite similar but a possible larger number of accessible and charged residual silanols was noticed on the wide-pore prototype material, which led to unpredicted small changes in selectivity and slightly broader peaks than expected. The peak shapes attained with the addition of 0.1% formic acid in the mobile phase remained acceptable for MS coupling, particularly for biomolecules of less than 6 kDa. It was found that one of the major issues with all of these silica-based monoliths is the possible poor recovery of large biomolecules (principally with monoclonal antibody fragments of more than 25 kDa).
Keywords Monoclonal antibodiesPeptidesProteinsProtein adsorptionSilica-based monolithWide-pore
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VUIGNIER, Karine et al. Comparison of various silica-based monoliths for the analysis of large biomolecules. In: Journal of separation science, 2013, vol. 36, n° 14, p. 2231-2243. doi: 10.1002/jssc.201300323 https://archive-ouverte.unige.ch/unige:29518

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Deposited on : 2013-09-02

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