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Lookup NU author(s): David Blinco,
Dr Glen Kemp,
Dr Achim Treumann
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The separation of intact proteins, including protein isoforms arising from various amino-acid modifications, employing a poly(styrene-co-divinylbenzene) monolithic capillary column in high-performance liquid chromatography coupled on-line to a time-of-flight mass spectrometer (MS) is described. Using a 250 mm × 0.2 mm monolithic capillary column high-sensitivity separations yielding peak capacities of >600 were achieved with a 2h linear gradient and formic acid added in the mobile phase as ion-pairing agent. The combination of high-resolution chromatography with high-accuracy MS allowed to distinguish protein isoforms that differ only in their oxidation and biotinylation state allowing the separation between structural isoforms. Finally, the potential to separate proteins isoforms due to glycosylation is discussed.
Author(s): Eeltink S, Wouters B, Desmet G, Ursem M, Blinco D, Kemp GD, Treumann A
Publication type: Article
Publication status: Published
Journal: Journal of Chromatography A
Print publication date: 21/06/2011
ISSN (print): 0021-9673
ISSN (electronic): 1873-3778
Publisher: Elsevier BV
PubMed id: 21741050
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