Challenges of glycoprotein analysis by microchip capillary gel electrophoresis.
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.
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.
Nicole
N
Engel
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.
Victor U
VU
Weiss
Agilent Technologies, Waldbronn, Germany.
Christian
C
Wenz
Agilent Technologies, Waldbronn, Germany.
Andreas
A
Rüfer
Agilent Technologies, Waldbronn, Germany.
Martin
M
Kratzmeier
Agilent Technologies, Waldbronn, Germany.
Susanne
S
Glück
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.
Martina
M
Marchetti-Deschmann
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.
Günter
G
Allmaier
D046209:Electrophoresis, Microchip / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D006023:Glycoproteins
Chip electrophoresis; Glycoprotein; Lab-on-a-chip; Laser-induced fluorescence; SDS-PAGE