Compositional Monosaccharide Analysis of Transgenic Corn Glycoproteins by HPLC with Fluorescence Detection and LC-MS with Sonic Spray Ionization

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.

Department(s)

Chemistry

Sponsor(s)

Monsanto Protein Tech

Keywords and Phrases

Glycoprotein; Monosaccharide; N Acetylneuraminic Acid; Chemistry; High Performance Liquid Chromatography; Mass Spectrometry; Methodology; Spectrofluorometry; Transgenic Plant; Chromatography, High Pressure Liquid; Glycoproteins; Monosaccharides; N-Acetylneuraminic Acid; Plants, Genetically Modified; Spectrometry, Fluorescence; Spectrometry, Mass, Secondary Ion; Zea Mays

International Standard Serial Number (ISSN)

0021-9665

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2007 Preston Publications, All rights reserved.

Publication Date

01 Apr 2007

Link to Full Text

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