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Title: Compositional monosaccharide analysis of transgenic corn glycoproteins by HPLC with fluorescence detection and LC-MS with sonic spray ionization
Author (s): Wang, Tongwen
Nam, Paul Ki-souk
Shi, Honglan
Ma, Yinfa
Department/Lab Affiliations: Chemistry
Environmental Research Center
Keywords: Fluorescence detection.
Sonic spray ionization.
Subject Terms: Glycoproteins.
Monosaccharides.
Issue Date: 2007
Publisher: Preston Publications
Citation: Wang, Tongwen, Nam, Paul Ki-souk, Shi, Honglan, and Ma, Yinfa, “Compositional Monosaccharide Analysis of Transgenic Corn Glycoproteins by HPLC with Fluorescence Detection and LC/MS with Sonic Spray Ionization”, Journal of Chromatographic Science, 2007, 45, 200-206.
Abstract: 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.
Type: Article - Journal
text
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titleCompositional monosaccharide analysis of transgenic corn glycoproteins by HPLC with fluorescence detection and LC-MS with sonic spray ionization
contributor.authorWang, Tongwen
contributor.authorNam, Paul Ki-souk
contributor.authorShi, Honglan
contributor.authorMa, Yinfa
contributor.deptlabChemistry
contributor.deptlabEnvironmental Research Center
contributor.sponsorMonsanto Protein Tech
subjectFluorescence detection.
subjectSonic spray ionization.
subject.LCSHGlycoproteins.
subject.LCSHMonosaccharides.
date.issued2007
publisherPreston Publications
identifier.citationWang, Tongwen, Nam, Paul Ki-souk, Shi, Honglan, and Ma, Yinfa, “Compositional Monosaccharide Analysis of Transgenic Corn Glycoproteins by HPLC with Fluorescence Detection and LC/MS with Sonic Spray Ionization”, Journal of Chromatographic Science, 2007, 45, 200-206.
identifier.pub.URI
http://www.ingentaconnect.com/content/pres/jcs/2007/00000045/00000004/art00006
description.abstractTransgenic 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.
typeArticle - Journal
type.DCMITypetext
type.statusFinal version
rightsThis material is presented to ensure timely dissemination of scholarly and technical work. Copyright and all rights therein are retained by authors or by other copyright holders. All persons copying this information are expected to adhere to the terms and constraints invoked by each author's copyright. In most cases, these works may not be reposted without the explicit permission of the copyright holder.
rights.URI
http://www.j-chrom-sci.com/authors.htm#Copyright
date.accessioned2007-04-11T17:00:48Z
date.available2007-12-17T20:41:10Z
identifier.persist.URI
http://scholarsmine.mst.edu/post_prints/CompositionalMonosaccharideAnalysis_09007dcc8042f1dc.html