Software Development for xCGE-LIF-Based Glycan Analysis

The analysis of protein glycosylation became of increasing importance in glycobiological research, as well as for bioprocess monitoring during development and production of biopharmaceuticals and vaccines in the recent past. In order to enhance and improve the existing glycoanalytical toolbox, automated high-throughput, high-resolution and highly sensitive but robust analysis methods with automated data processing and analysis are required. Multiplexed capillary gel electrophoresis with laser induced fluorescence detection (xCGE-LIF), a quite feasible electrokinetic separation technique using standard DNA sequencer equipment, has been developed for glycoprofiling of fluorescently labeled glycans. However, when it comes to data analysis bioinformatics software is still lacking for processing experimental data.

Graphical user interfaces (GUI) of the modular glycoanalysis software tool for HT data processing and evaluation.

© MPI Magdeburg (BPE)

Graphical user interfaces (GUI) of the modular glycoanalysis software tool for HT data processing and evaluation.

© MPI Magdeburg (BPE)

To fully exploit the potential of this method for high-performance glycoanalysis, data analysis has to be automated. In cooperation with the glyXera GmbH in Magdeburg/Germany, a novel modular software tool for data processing and analysis is developed and interfaced with a corresponding glyco-database. Using this software tool, "normalized" electropherograms of glycomoieties ("fingerprints"), generated by this glycoanalysis system, can be evaluated on two stages: "simple" qualitative and quantitative fingerprint comparison and/or structural elucidation of each of the glycocomponents of the samples.

Publications

Rapp, E.; Hennig, R.; Borowiak, M.; Kottler, R.; Reichl, U.: High-Throughput Glycosylation Pattern Analysis of Glycoproteins Utilizing a Multiplexing Capillary-DNA-Sequencer. In Glycoconjugate Journal, 28 (5 - Special Issue : XXI International Symposium on Glycoconjugates), p. 234. 21st International Symposium on Glycoconjugates - Glyco 21, Wien, Östereich, August 21, 2011 - August 26, 2011. (2011)

MPG.PuRe

DOI

Hennig, R.; Reichl, U.; Rapp, E.: A Software Tool for Automated High-Throughput Processing of CGE-LIF Based Glycoanalysis Data, Generated by a Multiplexing Capillary DNA Sequencer. In Glycoconjugate Journal, 28 (5 - Special Issue : XXI International Symposium on Glycoconjugates), p. 331. 21st International Symposium on Glycoconjugates (Glyco 21), Wien, Östereich, August 21, 2011 - August 26, 2011. (2011)

MPG.PuRe

DOI

Behne, A.; Muth, T.; Borowiak, M.; Reichl, U.; Rapp, E.: glyXalign: High-throughput migration time alignment preprocessing of electrophoretic data retrieved via multiplexed capillary gel electrophoresis with laser-induced fluorescence detection-based glycoprofiling. Electrophoresis 34 (16), pp. 2311 - 2315 (2013)

MPG.PuRe

DOI

post-print

Patents

[1] E Rapp, J Schwarzer, U Reichl, C Bohne: Method for automated high throughput identification of carbohydrates and carbohydrate mixture composition patterns as well as systems therefore. European patent: EP2533039 (A1) (2008).

[2] E Rapp, J Schwarzer, U Reichl, C Bohne: Method for automated high throughput identification of carbohydrates and carbohydrate mixture composition patterns as well as systems therefore. US patent: US2013075258 (A1) (2009).