Compositional monosaccharide analysis of transgenic corn glycoproteins by HPLC with fluorescence detection and LC-MS with sonic spray ionization.
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.
Wang
Tongwen
T
Department of Chemistry and Center for Environmental Science and Technology, University of Missouri-Rolla, Rolla, MO 65409, USA.
Nam
Paul Ki-Souk
PK
Shi
Honglan
H
Ma
Yinfa
Y
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis; D009005:Monosaccharides / Q000032:analysis*; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D030821:Plants, Genetically Modified / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*
D006023:Glycoproteins; D009005:Monosaccharides; D019158:N-Acetylneuraminic Acid