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ID 50974
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Author
Tabara, Ayumi
Kaneta, Takashi ORCID Kakenhi
Abstract
Here, we report a novel method consisting of capillary electrophoretic separation followed by two-color LIF detection with postcolumn derivatization. The method can be used to discriminate glycoproteins in a protein mixture containing both glycosylated and unglycosylated proteins. The detector permitted simultaneous measurements of two electropherograms obtained by 450 nm (diode laser) and 532 nm (Nd:YAG laser) lasers excited native proteins following postcolumn derivatization with naphthalene-2,3-dicarboxaldehyde and concanavalin A (Con A) labeled with tetramethylrhodamine (rhodamine-labeled Con A), respectively. So, a protein can be assigned as glycosylated if it shows a peak at the same migration time in both electropherograms. According to the proposed principle, in a single run we discriminated a glycosylated protein (thyroglobulin) from an unglycosylated protein (albumin) in the presence of rhodamine-labeled Con A. Because the methodology permits the simultaneous detection of native proteins and their complexes with a fluorescently labeled probe, it should have broad applicability to binding assays.
Keywords
Capillary electrophoresis
Concanavalin A
Glycoprotein
Postcolumn derivatization
Two-color laser-induced fluorescence
Published Date
2013-08
Publication Title
Electrophoresis
Volume
volume34
Issue
issue16
Publisher
WILEY-VCH
Start Page
2316
End Page
2322
ISSN
01730835
NCID
AA00179882
Content Type
Journal Article
language
英語
OAI-PMH Set
岡山大学
Copyright Holders
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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isVersionOf https://doi.org/10.1002/elps.201300149