Team Leader (BPA)

Dr. Erdmann Rapp
Dr. Erdmann Rapp
Phone: +49 391 6110 314
Fax: +49 391 6110 535
Room: S0.13

Researchers

Samanta Cajic, MPharm
Phone: +49 391 6110 334
Room: S0.09

Additional Information

Collaboration:
glyXera GmbH, Magdeburg/Germany

Supported by:
European Commission (EC) project: "High Glycan" in the European Union’s Seventh Framework Programm, Call: FP7-Health-F5-2011 (grant agreement no. 278535)
Started: 11/2011
http://www.highglycan.eu

Method Development for Multiplexing CGE-LIF Based Glycoanalysis

Header image 1380273611

Method Development for Multiplexing CGE-LIF Based Glycoanalysis

Motivation

In cooperation with the glyXera GmbH in Magdeburg/Germany, a method for HT/HP/HR glycoanalysis, based on multiplexed capillary gel electrophoresis with laser induced fluorescence detection (xCGE-LIF) utilizing a DNA-sequencer, is developed at the MPI. The application of this technique to glycoanalysis using instruments with up to 96 capillaries in parallel, results in massive reduction of the effective separation time per sample combined with an impressive sensitivity achieved due to LIF detection and therefore, shows high potential for HT/HP/HR glycoprofiling of glycoconjugates.

<small>Workflow of high-throughput glycosylation pattern analysis of glycoproteins utilizing a multiplexing capillary-DNA-sequencer.</small> Zoom Image
Workflow of high-throughput glycosylation pattern analysis of glycoproteins utilizing a multiplexing capillary-DNA-sequencer. [less]

Aim of the project

The aim of this project is to further investigate and to improve this innovative approach for use in various fields of application. Therefore, the sample preparation method and workflow are further optimized with respect to performance and feasibility regarding HT, and the separation parameters (e.g.: gel matrix composition, voltage or capillary length) are evaluated regarding HP and HR, to find the optimum for an automated separation for each of a set of different types of samples.

References

1.
Hennig, R.; Rapp, E.; Kottler, R.; Cajic, S.; Reichl, U.: N -Glycosylation Fingerprinting of Viral Glycoproteins by xCGE-LIF. In: Carbohydrate-Based Vaccines: Methods and Protocols, pp. 123 - 143. Springer, New York (2015)
2.
Borowiak, M.; Hennig, R.; Kottler, R.; Reichl, U.; Rapp, E.: Tailored methods for CGE-LIF based multiplexed high-throughput glycoanalysis with respect to sample characteristics. In: Glycoconjugate Journal, Vol. 28, p. 332. 21st International Symposium on Glycoconjugates - Glyco 21, Wien, Östereich, August 21, 2011 - August 26, 2011. (2011)
3.
Schwarzer, J.; Rapp, E.; Reichl, U.: N-Glycan Analysis by CGE-LIF – Profiling Influenza A Virus Hemagglutinin N-Glycosylation during Vaccine Production. Electrophoresis 29 (20), pp. 4203 - 4214 (2008)

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).

 
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