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Challenges of glycoprotein analysis by microchip capillary gel electrophoresis
Author(s) -
Engel Nicole,
Weiss Victor U.,
Wenz Christian,
Rüfer Andreas,
Kratzmeier Martin,
Glück Susanne,
MarchettiDeschmann Martina,
Allmaier Günter
Publication year - 2015
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400510
Subject(s) - analyte , chromatography , capillary electrophoresis , reproducibility , chemistry , glycan , sizing , glycosylation , glycoprotein , capillary action , gel electrophoresis , analytical chemistry (journal) , materials science , biochemistry , organic chemistry , composite material
Glycosylations severely influence a protein's biological and physicochemical properties. Five exemplary proteins with varying glycan moieties were chosen to establish molecular weight (MW) determination (sizing), quantitation, and sensitivity of detection for microchip capillary gel electrophoresis (MCGE). Although sizing showed increasing deviations from literature values (SDS‐PAGE or MALDI‐MS) with a concomitant higher degree of analyte glycosylation, the reproducibility of MW determination and accuracy of quantitation with high sensitivity and reliability were demonstrated. Additionally, speed of analysis together with the low level of analyte consumption render MCGE attractive as an alternative to conventional SDS‐PAGE.