Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume 45,
Number 4, April 2007, pp.200-206
Compositional Monosaccharide Analysis of Transgenic
Corn Glycoproteins by HPLC with Fluorescence Detection and LC–MS
with Sonic Spray Ionization
Tongwen Wang, Paul Ki-souk Nam, Honglan Shi, and
Yinfa Ma
Department of Chemistry and Center for Environmental Science and
Technology, University of Missouri-Rolla, Rolla, MO, 65409
Transgenic corn offers an attractive, cost-effective
means for the large-scale production of engineered glycoproteins
suitable for pharmaceutical purposes. A glycoprotein expressed
in transgenic corn theoretically should not contain glycans because
glycosylation sites have been genetically altered. A sensitive
and reliable analytical method is developed to investigate this
particular protein for the presence of glycans by monitoring the
monosaccharide composition. Identification and quantitation of
low-level monosaccharides in the glycoprotein hydrolyzate are
accomplished by derivatization prior to high-performance liquid
chromatography (HPLC)–fluorescence and liquid chromatography
(LC)–sonic spray ionization (SSI)-mass spectrometry (MS)
analyses. LC–SSI-MS is used to confirm the results from
HPLC–fluorescence analysis and to positively identify the
compositional monosaccharides. Glucosamine, glucose, mannose,
arabinose, xylose, and sialic acid are found in the transgenic
corn derived glycoprotein at less than one moiety per protein,
which indicates heterogeneity of the particular glycoprotein.
In addition to the compositional analysis of low-level monosaccharides
in glycoprotein by HPLC–fluorescence, the utility of SSI
for the LC–MS analysis of derivatized monosaccharides is
demonstrated in this paper.
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