In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins
In this study, we present a new approach for in‐capillary fluorescent labeling of N‐glycans prior to their analysis with CE coupled with laser‐induced fluorescent detection. This integrated approach allows using a CE capillary as a microreactor to perform several steps required for labeling glycans with 8‐aminopyrene‐1,3,6 trisulfonic acid and at the same time as a separation channel for CE of fluorescently labeled glycans. This could be achieved through careful optimization of all different steps, including sequential injections of fluorescent dye and glycan plugs, mixing by transverse diffusion of laminar flow profiles, incubation in a thermostatic zone, and finally separation and detection with CE. Such a complex sample treatment protocol for glycan labeling that is feasible thus far only in batchwise mode can now be converted into an automated and integrated protocol. Our approach was applied successfully to analyze fluorescently labeled N‐linked oligosaccharides released from human immunoglobulin G and rituximab, a monoclonal antibody used for cancer treatment. We demonstrated the superiority of this in‐capillary approach over the conventional in‐tube protocol, with fourfold less reagent consumption and full automation without remarkable degradation of the glycan separation profile obtained by capillary electrophoresis.
- Standort
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Deutsche Nationalbibliothek Frankfurt am Main
- Umfang
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Online-Ressource
- Sprache
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Englisch
- Erschienen in
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In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins ; day:22 ; month:07 ; year:2022 ; extent:10
Journal of separation science ; (22.07.2022) (gesamt 10)
- Urheber
- DOI
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10.1002/jssc.202200340
- URN
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urn:nbn:de:101:1-2022072315072157827410
- Rechteinformation
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Letzte Aktualisierung
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28.03.8150, 04:39 MEZ
Datenpartner
Deutsche Nationalbibliothek. Bei Fragen zum Objekt wenden Sie sich bitte an den Datenpartner.
Beteiligte
- Yang, Bin
- Mai, Thanh Duc
- Tran, Nguyet Thuy
- Taverna, Myriam
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