Oral Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

glyXboxCE: A powerful tool in the glycoanalytical toolbox - improving biologics development and biomarker discovery for personalized diagnostics (#137)

Erdmann Rapp 1 2 , René Hennig 2 , Samanta Čajić 1 , Alexander Behne 2 , Robert Kottler 2 , Udo Reichl 1 3
  1. Max-Planck-Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany
  2. glyXera GmbH, Magdeburg, Germany
  3. Otto-von-Guericke University, Magdeburg, Germany

Glycomics is a rapidly emerging field that can be viewed as a complement to other „omics“ approaches. Hence, there is a dramatic increase in the demand for analytical tools and specific databases in glycobiology, respectively, glycobiotechnology. In order to enhance and improve the comparatively small existing glycoanalytical toolbox, automated, sensitive, reliable, high-throughput and high-resolution analysis methods including automated data evaluation are required. Our glycoanalysis approach, based on multiplexed capillary gelelectrophoresis with laser induced fluorescence detection (xCGE-LIF), shows high potential for high-performance analysis of glycoconjugates, as it allows fully automated, highly sensitive, instrument-, lab- and operator-independent "real" high-throughput glycoanalysis. This novel modular glycoanalysis system “glyXboxTM” and its application to different fields with respect to sample preparation, separation and data analysis is presented. First, an optimized modular sample preparation workflow is presented with respect to performance and feasibility regarding high-throughput analytics. Second, parallel sample-measurement is shown to result in massive reduction of the effective run-time per sample. Third, automated data analysis with a newly developed modular software-tool “glyXtoolCE” for data processing and data analysis is demonstrated that involves integration of a corresponding oligosaccharide-database. Using this high-performance xCGE-LIF based glycoanalysis system, the generated “normalized” electropherograms of glycomoieties (“fingerprints”) can be evaluated on three levels: (1) “simple” qualitative and quantitative pattern comparison (“glycofingerprinting”), (2) identification of compounds in complex mixtures via database matching (“standard glycoprofiling”), and (3) extended structural analysis using exoglycosidase sequencing in combination with xCGE-LIF based glycoprofiling with repeated xCGE-LIF-based glycoprofiling (“extended glycoprofiling”), including linkage analysis. The broad applicability of the system is demonstrated for different types of glycosamples and different application fields - from biologics for biopharma to body fluids for personalized diagnostics.

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