Identification of glycoconjugates in the urine of a patient with congenital disorder of glycosylation by high-resolution mass spectrometry

Authors

  • Sergey Y. Vakhrushev,

    1. Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Münster, Germany
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  • Michael Mormann,

    1. Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Münster, Germany
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  • Jasna Peter-Katalinić Professor

    Corresponding author
    1. Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Münster, Germany
    • Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Robert-Koch-Str. 1, D-48149 Münster, Germany Fax: +49-251-8355140
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Abstract

More than 150 molecular species were detected in a single glycoconjugate fraction obtained from urine of a congenital disorders of glycosylation (CDG) patient by use of high-resolution FT-ICR MS. With respect to its high-mass accuracy and resolving power, FT-ICR MS represents an ideal tool for analysis of single components in complex glycoconjugate mixtures obtained from body fluids. The presence of overlapping nearly isobaric ionic species in glycoconjugate mixtures obtained from CDG patient's urine was postulated from fragmentation data of several precursor ions obtained by nanoESI Q-TOF CID. Their existence was confirmed by high-resolution/high-mass accuracy FT-ICR MS detection. High-resolution FT-ICR mass spectra can, therefore, be generally considered for glycoscreening of complex mixture samples in a single stage. From the accurate molecular ion mass determinations the composition of glycoconjugate species can be identified. Particular enhancement of identification is offered by computer-assisted calculations in combination with monosaccharide building block analysis, which can be extended by considerations of non-carbohydrate modifications, such as amino acids, phosphates and sulfates. Taking advantage of this strategy, the number of compositions assigned to mass peaks was significantly increased in a fraction obtained from urine by size exclusion and anion exchange chromatography.

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