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Abstract:
Glycomics is a rapidly emerging field that can be viewed as a continuation of the genomic and proteomic era. Hence, there is a dynamic increase in the demand for sophisticated databases and smart analytical tools in glycobiology, respectively, glycobiotechnology. In order to enhance and improve the comparatively small existing glycoanalytical toolbox a fully automated (high-throughput (HTP) and high-resolution (HR)) analysis method with a fully automated data evaluation is required. Besides several mass spectrometry and liquid chromatography based analysis techniques, electromigrative separation techniques for the analysis of oligosaccharides became apparent during the recent past. Especially, capillary gel electrophoresis with laser induced fluorescence detection (CGE-LIF) as one feasible electromigrative separation technique - using standard DNA sequencer equipment - has been established for HTP glycoprofiling of APTS-labeled glycans [1]. The application of this technique using instruments with up to 96 capillaries in parallel to glycoanalysis, results in massive reduction of the effective separation time per sample combined with an impressive sensitivity achieved due to LIF detection [2]. Due to the lack of appropriated software for data analysis, the MATLAB® based software tool “glyXtool” was developed, now significantly reducing the all-over analysis time per sample. A graphical user interface makes this tailor made glycoanalysis software-tool for automated data-processing easy to handle also by non-experts. Thereby, glyXtool provides automated background adjustment, raw data smoothing, migration time normalization to an internal standard, peak picking and peak integration in HTP. All this, combined with glyXtools intrinsic sample comparison function, a fully automated peak annotation and an interface to a continuously growing oligosaccharide database, makes multiplexed CGE-LIF to a powerful glycoanalysis tool. This is in contrast to currently prevailing methods, where multiplexing with respect to HTP is highly cost and lab-space intensive and ties up a lot of manpower and experts hands-on-time.
[1] Schwarzer J, et al., Electrophoresis, 2008, 29, 4203-4214.
[2] Ruhaak LR, et al., Journal of Proteome Research, 2010, 9,6655-6664.
