8 Key Areas of Research and Development – Advancements in Healthcare Will Be Sweet Systematic Glycoanalytics – Unveiling Complex Glycan Structures and Functions and Identifying New Biomarkers Glycoanalytics build the basis for all glycobiotechnology applications. Several technologies are applied to increase knowledge about glycans, its constituents and structure as well as its innumerable functions in health and disease. Beside their functions in energy metabolism, carbohydrates are constituents of glycoproteins, glycolipids, proteoglycans (glycosaminoglycans) and glycosphingolipids, and many diseases and disorders can be linked to alterations in glycosylation. To give one example of the great variety of glycans: linking three different monosaccharides can lead theoretically to over 20,000 different structures, which is incredible when one considers that linking three amino acids produces just 27 tripeptides. Almost all regional academic institutions and several companies committed to the development of glycoengineered therapies or vaccines employ glycoanalytical techniques as part of their research efforts and to confirm their results. These include particularly the companies Glycotope GmbH, ProBioGen AG, Octapharma Biopharmaceuticals GmbH and UGA Biopharma GmbH. One of the major players is the Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité – Universitätsmedizin Berlin. The working group of Véronique Blanchard aims at the systematical analysis of the glycome in the context of physiological mechanisms in human health and disease. The group is able to analyze glycans from human sources, animal models and even plants. Several technical devices are used in the laboratory to integrate different technologies such as mass spectrometry (including Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry, MALDI-TOF MS), capillary electrophoresis and 2D gel electrophoresis, or other chromatographic techniques, namely high performance liquid chromatography (HPLC) and high-pH anion-exchange chromatography (HPAEC). The group uses these different techniques to unravel the whole glycome: N/O-glycans, glycosaminoglycans and glycosphingolipids. Kevin Pagel from the Institute of Chemistry and Biochemistry, Department of Biology, Chemistry, Pharmacy, at the Freie Universität Berlin addresses a tricky challenge of glycoanalytics. Carbohydrates are often branched and have a complex regio- and stereochemistry. This in turn leads to a vast number of possible isomers of identical atomic composition and mass, which are difficult to distinguish using established analytical tools. To solve this problem, Kevin Pagel uses a combination of ion mobility-spectrometry and mass spectrometry (IM–MS), a very promising new tool in glycan analysis that has emerged in recent years. Additionally, he applies cold-ion spectroscopy, a method that provides highly diagnostic absorption features for oligosaccharides, resulting in a spectral fingerprint that is unique for each oligosaccharide. The unprecedented resolution of cold-ion spectroscopy coupled with tandem MS may render this the key technology to unravel complex glycomes. “Protein glycosylation plays a crucial role in essential biological processes, e.g., immune response, cell differentiation and development, cell-cell interaction and signal transduction. Alterations in glycosylation are linked to vast numbers of diseases and disorders. Systematic knowledge of disease-related changes of glycosylation as well as their integration in genomic and proteomic data allows for a new understanding of biomedical mechanisms and thus opens up new possibilities for diagnostics, therapeutic approaches and prevention. Our working group, aiming at the systematical analysis of the glycome, is part of the excellent basic research environment based in the area Berlin-Brandenburg. Additionally, key players from the industry are also present in the region, meaning that paths for joint R & D projects are particularly short.” Véronique Blanchard Head Department Glycodesign and Glycoanalytics, Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité – Universitätsmedizin Berlin
Key Areas of Research and Development – Advancements in Healthcare Will Be Sweet 9 Glycan-based biomarkers leverage a new dimension of diagnostics Changes in glycosylation are associated with many diseases and have been documented for several disease conditions such as cardiovascular, metabolic, infectious and inflammatory diseases as well as for some hereditary disorders. Tremendous research efforts all over the world focus on exploring altered glycosylation in various malignancies as tumor-related glycosylation patterns can provide valuable diagnostic information on the type of cancer, staging, progression and the patient’s prognosis. The identification and evaluation of glycan-based biomarkers represents a prominent field of research in Blanchard’s lab. One of the dedicated research projects – together with the Department of Gynecology of the Charité – focuses on serum glycome profiling to identify tumor markers for the diagnosis of epithelial ovarian cancer (EOC). Common tumor markers for this type of cancer show poor specificity; therefore, better biomarkers are needed. The group was able to develop a GLYCOV index combining upregulated structures that were fucosylated and sialylated, and a group of downregulated structures that were of the high-mannose type. The research results indicate that serum glycome profiling could contribute to an improved diagnosis of ovarian cancer as GLYCOV could differentiate between early stage EOC and benign ovarian growths. Blanchard’s group is also working towards the next generation of stem cell (SC) markers. The scientists work with adult multipotent bone marrow SCs and embryonal SCs in collaboration with the Berlin-Brandenburg Center for Regenerative Therapies (BCRT). SCs have the potential to develop into several tissues and are ideal candidates for applications in regenerative medicine. The group has identified key glycan structures in SCs that are promising candidate markers to detect and distinguish SCs from their adipogenic progeny. These and other projects reveal the potential of glycan-based biomarkers as determinants of physiological processes in health and disease. Furthermore, new glycan-based diagnostic methods enable more personalized therapy approaches and improved patient care.