Donnerstag, 26. April 2018 I 13:00 – 17:00 Uhr
Fraunhofer IZI-BB I Am Mühlenberg 13 I 14476 Potsdam

Das Ziel des glyconetBB ist es, Synergien zwischen Wissenschaftlern und Unternehmern auf den Gebieten der Glykowissenschaft und Glykobiotechnologie zu identifizieren, Kooperationen zu initiieren sowie die Entwicklung innovativer Produkte für die Gesundheitswirtschaft in der Hauptstadtregion zu begleiten.
Im ersten Statusseminar möchten wir Ihnen das glyconetBB mit seinen Mitgliedern und Tätigkeitsschwerpunkten vorstellen. Es bietet die Möglichkeit, Netzwerkpartner kennenzulernen und mit ihnen zukünftige Projekte zu diskutieren.

PROGRAMM hier herunterladen



Bestätigte Referenten:

Prof. Dr. Rudolf Tauber (Charité – Universitätsmedizin Berlin)
Prof. Dr. Lothar Elling (RWTH Aachen, Helmholtz-Institut für Biomedizinische Technik)
Prof. Dr. Stephan Hinderlich (Beuth Hochschule für Technik Berlin)
Dr. Sven Bahrke (glycotope GmbH)
Dr. Erdmann Rapp (glyXera GmbH)
Dr. Ruben R. Rosencrantz (Fraunhofer IAP)
Dr. Marlitt Stech (Fraunhofer IZI-BB)
Dr. Nathalie Veith (UGA Biopharma GmbH)


Wir hoffen, Ihnen ein abwechslungsreiches Programm bieten zu können,
das Ihr Interesse weckt, und freuen uns auf Ihr Kommen.

Aufgrund der limitierten Teilnehmerzahl wird um Anmeldung gebeten
info@glyconet.de I https://glyconet.eventbrite.de I 0331-97921972

Die Teilnahme ist kostenlos.

Winter School 2017 glyconet e.V.

Successfully finished Winterschool „Moderne Methoden zur Synthese und Analyse von Glykoproteinen“ des glyconet Berlin Brandenburg e.V.

Winterschool 2017

Winter School 2017 glyconet

Das glyconet Berlin-Brandenburg (BB) ist ein Zusammenschluss von Firmen und Institutionen zur Förderung innovativer Forschungsansätze in der Zuckerforschung. Einen Einblick in die vielfältige Welt der Zuckerforschung ermöglichte das glyconet BB jetzt für Nachwuchswissenschaftler. Studenten verschiedener Institute aus Berlin und Brandenburg konnten Ende Januar 2017 bei der einwöchigen Winterschool Theorie und Praxis der Glykoforschung erleben. Als Dozenten konnten ausgewiesene Experten auf Ihrem jeweiligen Fachgebiet gewonnen werden. Diese erläuterten in den Räumlichkeiten des Fraunhofer-Institut für Zelltherapie und Immunologie IZI in Seminaren Themen wie Glykobiologie, Glykan Analytik, Antikörper Herstellung, zellfreier Glykoproteinsynthese und die Vielseitigkeit von Glykanen in biofunktionalen Materialien. Außerdem konnten die Teilnehmer praktische Versuche unter anderem zur Glykan Analytik und der zellfreien Synthese von Glykoproteinen im Labor selbst durchführen. Zum Abschluss der Winterschool wurde allen Teilnehmern der Besuch der Tagung GlycoBioTec 2017 in Berlin ermöglicht. Hier konnten weitere Disziplinen der Glykowissenschaften entdeckt und erste Kontakte zu anderen Forschern und Firmen geknüpft werden. Studenten und Dozenten waren sich am Ende einig, dass die Veranstaltung ein voller Erfolg war. Durch den Einblick in dieses spannende facettenreiche Forschungsfeld können angehende Wissenschaftler für die Glykowissenschaften begeistert werden. Somit stellt die Winterschool ein wertvolles Instrument zur zukünftigen Standortstärkung in Berlin/Brandenburg dar. Die Winterschool wird nun jedes Jahr in einem der Labore der glyconet BB Mitglieder durchgeführt.

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Glyconet Berlin Brandenburg e.V.

Glycoscience and glycobiotechnology opening a very fast growing market. The network is a platform to cooperate and interlink partners and arises opportunities to easily develop innovative products in the glycoscience area.

„glyconet BB“ is a network that supports research in the innovative glycobiotechnology and in the development of newly synthetic analytic and therapeutics for medicine engineering and life science. The main goal is to strengthen and visualize skills and education in the previous mentioned areas of life science.

Berlin Brandenburg has a unique value chain in glycobiotechnology and is now able to recruit partners from small and middle industry company as well as research institutes in hole germany for this cross-sectional technology.

Dr. Stefan Kubick, Fraunhofer IZI-BB

Dr. Stefan Kubick heads the department “Cell-free and Cell-based Bioproduction” at the Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses Potsdam-Golm. His laboratory exploits cell-free protein synthesis as a versatile tool for functional genomics, e.g. cell-free synthesis of membrane proteins and glycoproteins, as well as chip-based protein synthesis and translational regulation. Dr. Kubick is also a lecturer at the Freie Universität Berlin and the University of Potsdam. He is an affiliate of the Technical University of Berlin and Lecturer at the University of Applied Sciences, Berlin.

As a chairman of glyconet Berlin Brandenburg, he actively works on establishing the region as a center of excellence in glycobiology.

Dr. Kubick introduced us to the newest developments in the cell-free protein synthesis and adressed the possibilities of its application in the chemistry of sugars.

In the 1960s, Marshall Nirenberg and Heinrich Matthaei pioneered the use of prokaryotic enzyme extracts to characterize a range of essential factors in protein synthesis. One result of their famous “PolyU” experiment was to demonstrate the role of a ‘template RNA’ in protein synthesis – a milestone on the road to the ultimate discovery of the base triplet as the coding unit for amino acids. At the end of the 1980s, it was possible to significantly boost the capabilities of cell lysates, thanks especially to the work of Alexander Spirin, finally enabling the synthesis of larger quantities of proteins in cell-free systems. The “Continuous-Exchange Cell-Free” process patented by Spirin was based on a small bioreactor with a synthesis chamber, coupled to a reservoir tank that, via a dialysis membrane, continuously supplied the reaction with fresh consumables such as ATP and GTP as sources of energy, plus all of the necessary amino acids as building blocks for the protein synthesis itself. Small-molecule inhibitors were also removed from the synthesis chamber, as their increasing concentration disrupted the process. Left in the reaction chamber were the coding nucleic acids and the ribosomal machinery for protein synthesis, plus the gradually accumulating, newly synthesized proteins. This technology was the first to offer continuous cell-free synthesis of proteins over a period of up to 24 hours, which represented a significant improvement in protein output compared to other systems. Cell-free protein synthesis systems, also termed in vitro translation systems, expand the diversity of protein expression in a remarkable way. The intense development and analysis of pro- and eukaryotic in vitro translation systems are permanently growing fields of research. Open cell-free systems are designed for the synthesis of complex membrane proteins, glycoproteins and cytosolic proteins with the addition of posttranslational modifications onto the growing nascent polypeptide chain. The development of translationaly active cell lysates into highly productive systems, the generation and optimization of particular DNA and RNA templates for different cell-free systems and the optimization of reaction conditions in newly developed in vitro translation systems are part of our current research investigations.

Cell-free systems based on E. coli lysates utilise batch based and dialysis based procedures to deliver protein yields up to 5 mg/ml. Improvements here have focused primarily on ensuring high protein content in a soluble and functional form. In contrast to the conventional expression of proteins in cells and bacteria, the open, cell-free systems permit the configuration of the ionic milieu, the reaction temperature and the dosing of mild detergents, as well as folding helpers (chaperones) even during protein synthesis. Those proteins, termed “difficult-to-express proteins”, which to date have not been successfully expressed or which exhibit cytotoxic effects and kill off the host cell, can now be readily expressed by deploying an efficient template with a protein-specific supplementation of the translation reaction in automated, highly parallel cell-free systems. The expression of functionally active eukaryotic proteins requires not only the coordinated interaction of the protein expression machinery but also the integrity of all co- and post-translationally protein modifying components. If human genes are to be manufactured in a modified and simultaneously active form, this is best done in a system that most closely corresponds to the natural environment of these proteins – and at the point in time of synthesis. For this reason, there is large-scale use of cultivated mammalian cells – and human cell lines in particular – for the human-identical manufacturing of pharmacologically relevant proteins in eukaryotic cell-free systems. The decisive advantages of cell-free protein synthesis in eukaryotic in vitro translation systems, such as the high-throughput synthesis of continuously active, cytotoxic membrane proteins, can be combined with biochip-based, impedimetric and electrophysiological functional analysis. A typical yield for a protein synthesized in a eukaryotic cell-free system is 10 to 100 µg per ml depending on the individual protein to be produced.

Cultivated eukaryotic cell lines possess the necessary subcellular structures to perform protein glycosylation and the enzymatic activities found within them are a prerequisite to perform these protein modifications. They therefore constitute an ideal starting substrate for the manufacture of eukaryotic in vitro translation systems. In this way, the general advantages of cell-free protein synthesis– such as the possibility of synthesising cytotoxic and labelled proteins – are combined with a rapid and easy to manage expression procedure for functionally active eukaryotic proteins. Due to the procedure utilised for the homogenisation of eukaryotic cells, intracellular, vesicular structures are retained in a functionally active form in the lysate. These microsomal elements are deployed for co-translational translocation and the immobilisation of cell-free synthesised membrane proteins on chip surfaces. The functional integrity of these subcellular components can be demonstrated by means of signal peptide cleavage, as well as the glycosylation of cell-free synthesised proteins. This enormous potential of eukaryotic in vitro translation systems is also utilised in the manufacture of antibody fragments, which involves the accumulation of defined,posttranslationally modified and fluorescence- labelled antibody fragments in proteoliposomes.

Many issues, such as lysate preparation, optimization of reaction conditions, and the design of the expression vector are taken into consideration for cell-free synthesis of functional proteins. Particularly, the latter is a crucial aspect for high-yield in vitro protein production. The performance of the cell-free systems is evaluated by synthesizing functional cytosolic proteins, posttranslationally modified proteins and membrane-spanning proteins. To monitor protein quality and quantity, reaction mixtures are supplemented with 14C labeled leucine. De novo synthesized proteins are routinely analyzed by autoradiography and radioactively labeled proteins are visualized using a phosphorimager system. Protein yield is determined by hot TCA precipitation. Additionally, detailed analysis is performed by mass spectrometry. Finally, cell-free produced ion channels for example, are analyzed by electrophysiological measurements.

Various cell-free protein synthesis systems based on cultured insect cells as well as cultured mammalian cells have been developed in the past decade in order to address the synthesis of posttranslationally modified proteins. The use of recently established lysates derived from these cultured insect cells and mammalian cells is preferred, as these extracts already contain endogenous microsomes derived from the endoplasmic reticulum of the cultured cells and these cell-free protein synthesis platforms have the potential to perform signal sequence processing and N-linked glycosylation. An emerging number of publications illustrates the application of these cell-free systems for the synthesis of co- and post-translationally modified proteins. Additionally, target proteins could be engineered in a desired manner by making use of the accessible nature of the cell-free protein synthesis reaction. In this context, bioorthogonal systems provide a promising tool for the incorporation of chemoselective reactive amino acids into synthesized proteins at defined positions by expanding the genetic code. These amino acids can subsequently be modified in a desired manner by the addition of a corresponding reaction partner, e.g. chemically synthesized glyco polymers, resulting in novel characteristics of the target protein. Several bioorthogonal and chemoselective reactions have been identified and applied in cell-free systems in recent years and this technology will obviously expand the number of applications of cell-free protein synthesis in the field of pharmaceutical research and development.

High-throughput protein expression platforms are becoming increasingly important. A unique and exciting development is the potential to enable the production of personalized medicines. Cell-free systems in particular, have many advantages for meeting this need. Cell-free expression methodology is well suited for pharmaceutical protein expression and engineering and will probably become more commonly used in the future. In the past decade, substantial progress has been made in the development of novel mammalian-based cell-free protein synthesis systems. However, there are still limitations which are currently in the focus of research and developmental efforts. Ongoing challenges in the field of mammalian-based cell-free protein synthesis are the optimization of already established platforms and the development of novel systems in order to enable even higher protein yields and lower manufacturing costs while facilitating the synthesis of a huge number of biologically active target proteins. By applying a combination of cell-free protein synthesis, high-throughput screening technologies and protein engineering, the chances of obtaining a therapeutic protein with optimal properties such as efficacy, stability and solubility, can only be increased. In this way, cell-free protein synthesis will be a true alternative to cell based protein expression systems currently used in industrial applications.

‘‘Lab-on-chip“ technologies are believed to be one of the mainstream technologies within the next centuries and high-throughput multiplex immunoassays that measure hundreds of proteins in complex biological matrices in parallel have become significant tools for quantitative proteomics studies, diagnostic discovery, and biomarker-assisted drug development. In this context cell-free protein synthesis is an essential prerequisite for the „just-in-time on-chip synthesis“ of biomarkers and the development of label-free assays based on these freshly synthesized active proteins.

While used for decades as a foundational research tool for understanding transcription and translation, recent advances have made possible cost-effective microscale to manufacturing scale cell-free synthesis of complex proteins. Protein yields exceed grams protein produced per liter reaction volume, batch reactions last for multiple hours, costs have been reduced orders of magnitude and reaction scale has reached the 100-liter milestone. Recent advances have inspired new applications in the synthesis of protein libraries for functional genomics, the production of personalized medicines, and the expression of tailor-made biological recognition elements including membrane-bound enzymes and receptors.

Dr. Holger Eickhoff, Scienion AG

During the last 15 years at the top of the Scienion AG, Dr. Holger Eickhoff experienced highs and lows of the biotech start-up landscape. But the onetime competitive swimmer pulled through adjusting his business along the ever changing market. Today, he leads one of the most promising biotech companies in the region, gradually expanding to the global market.

During SCIENION’s foundation it was most helpful to have the Max Planck Society (Max-Planck Gesellschaft (MPG)) as a reliable partner. More than 15 years ago the MPG had already had established technology transfer platform that professionally supported the transfer of new ideas coming out of basic research to the market, creating a business opportunity. Locally we benefitted from the BioTOP network (today healthcapital.de), since we both operate at the interface between science and business. However, in my opinion one approach is still missing: not every invention or business idea should lead to a distinct start-up company. It should be rather evaluated if a new business can be incorporated in existing companies. This approach is still lacking or is, at least, underrepresented.

What held us back for the first two-three years in company development was actually that we had been too innovative and didn’t find users for our products.

We see a chance for better networking between the regional players and checking of options for joint projects at the same time establishing an initial, local customer base. Our particular interest aims at promoting the benefits of novel array-based diagnostics utilizing glycan structures. We expect to learn more about this new market through the glyconet platform.

The chances of glycosciences have been well recognized within the scientific community. However, we need more research and development.  From a practical point of view, we need large, well-characterized glyco libraries in adequate amounts for R&D to advance glycomics. The integration of glyconet activities into national and international initiatives would definitely bring benefits!

We have an excellent and very broad expertise in the region – this is true for both: companies and research institutions. The glycoscience community in Berlin and Brandenburg covers many areas: from therapeutics, vaccines and diagnostics via nutrition to material sciences.

It is important to have in mind that glycosciences represent a cross-sectional technology and it will give us a competitive edge only if we successfully bundle our top expertise across companies and academia!