Prof. Dr. Stefan Kalkhof is the head of the HAW Coburg group „Instrumental Bioanalysis“ as well as the Fraunhofer research unit „Protein Biomarker“ of the devision Therapy Validation, Fraunhofer Institute for cell therapy and immunology IZI.
The Fraunhofer team focus on the identification and validation of protein biomarker as well as the development and validation of diagnostic single and multiplex assays (e.g. MS, ELISA, Luminex).
The research group at Coburg University uses spectroscopic (FT-IR, UV-VIS) and mass spectrometric approaches (ESI-LC-MS, MALDI-MS) to study proteins (quantitation, modification, structure) and its interaction with low and high molecular weight interaction partners (drugs, polymers, carbohydrates).
Development and characterization of innovative implant materials
Due to demographic development, the number of patients with bone or soft tissue defects as well as chronic wounds increases. Thus, novel functionalized biomaterials and implants as well as novel methods for implant production are required to improve bone and tissue regeneration or substitution. To efficiently guide these developments, appropriate bio analytical approaches are needed, which provide comprehensive and detailed information concerning the wound region and the healing process based on a minimal invasive sampling procedure. In cooperation with partners from clinics, university and companies these methods are developed and applied.
Reference: Förster, Y., Schmidt, J. R., Wissenbach, D.K., Pfeiffer, S. E. M., Baumann, S., Hofbauer, L. C., Bergen, M. von, Kalkhof, S., and Rammelt, S., “Microdialysis Sampling from Wound Fluids Enables Quantitative Assessment of Cytokines, Proteins, and Metabolites Reveals Bone Defect-Specific Molecular Profiles,” PloSone, V.11, No. 7, 2016, e0159580.
Schmidt JR, Vogel S, Moeller S, Kalkhof S, Schubert K, von Bergen M, Hempel U. “Sulfated hyaluronic acid and dexamethasone possess a synergistic potential in the differentiation of osteoblasts from human bone marrow stromal cells” J Cell Biochem. 2018 Nov 28. doi:10.1002/jcb.28158
Mode of action of toxic compounds
According the European guideline REACH chemicals need to be tested comprehensively for environmental and health effects. Are there subtoxic effects? Which molecular mechanisms are affected? Are there additive or even synergistic effects? These questions are addressed in a project with the federal institute of risk assessment (BFR) and the Helmholtz Centre for environmental research – UFZ using toxico proteomics. In cooperation with Prof. Nolls group as well as with Fraunhofer the developed methods are applied in further projects.
Reference: Wewering, F., Jouy, F., Caliskan, S., Kalkhof, S., Bergen, M. von, Luch, A., and Zellmer, S., “Hepaticco-cultures in vitro reveal suitable to detect Nrf2-mediated oxidative stress responses on the bladder carcinogen o-anisidine,” Toxicology in vitro : an international journal published in association with BIBRA, V.40, 2017, pp. 153–160.
Steve U. Ayobahan, Elke Eilebrecht, Matthias Kotthoff, Lisa Baumann, Sebastian Eilebrecht, Matthias Teigeler, Henner Hollert, Stefan Kalkhof, Christoph Schäfers “A combined FSTRA-shotgun proteomics approach to identify molecular changes in zebrafish upon chemical exposure” Sci Rep. 2019; 9: 6599. Published online 2019 Apr 29
Diagnosis of kidney diseases A pathologically increased abundance of a monoclonal antibody of the gamma fraction of serum proteins is called monoclonal Gammopathy. For differentiation and a targeted therapy an accurate identification and characterization of this accumulation protein is required. In combination imaging MALDI mass spectrometry of proteins (Prof. Andreas Römpp, University of Bayreuth) and quantitative proteome analysis (Prof. Stefan Kalkhof, University of Coburg) and immunohistopathology (Frau Prof. Kerstin Amann, University of Erlangen) these challenge is addressed in order to improve diagnostics and molecular understanding of this and other kidney diseases.
Methods for evidence based on health promotion
As part of the project „Gesundheit messen“ in the work package headed by Prof. Dr. Janosch Hildebrand and Prof. Dr. Stefan Kalkhof methods to measure the hydration status of patients with non-accute exsiccosis will be developed and applied on a molecular level. Thus stratification of health promotion strategies will be examined in terms of efficiency.
Structure elucidation of protein complexes e.g. TSH/TSH receptor interaction.
The TSH receptor (thyrotropin receptor) is a G-Protein coupled receptor of the thyroid, which is activated by thyrotropin. This activation triggers the formation of thyroid hormones.
A malfunction can cause e.g. Morbus Basedow.
The goal of this cooperation project with the groups of Prof. Dr. Jens Meiler (Vanderbilt Universität, USA) and Prof. Dr. Ralf Paschke (University Calgary, Canada) is the elucidation of the protein structure to pave the way for the development of activing and blocking drugs.
Reference: Schaarschmidt, J., Nagel, M. B. M., Huth, S., Jaeschke, H., Moretti, R., Hintze, V., Bergen, M. von, Kalkhof, S., Meiler, J., and Paschke, R., “Rearrangement of the Extracellular Domain/Extracellular Loop 1 Interface Is Critical for Thyrotropin Receptor Activation,” The Journal of biological chemistry, V.291, No. 27, 2016, pp. 14095–14108.
Biotechnical production of stable isotope marked (SI) amino acids and amino acid precursors for markings in mammalian cell cultures (funded by Bundesministerium für Wirtschaft und Energie aufgrund eines Beschlusses des Deutschen Bundestages)
Marking with stable isotopes (13C, 15N) does not result in functional changes in the labeled biomolecules in general, so this significant marking method became more and more accepted in the fields of biology, pharmacy and medicine and induces an increasing requirement of applications. Within the framework of this ZIM project, industrially applicable biotechnological production methods for SI amino acids or their precursor molecules will be developed in cooperation with eminent partners from industry and science. Simultaneously, the metabolic marking methods for various applications in NMR spectroscopy and proteomics are further developed and refined.
Ecotoxicological Testing of Biocidal Building Materials by conventional und systems biology approaches (funded by the Bavarian State Ministry of the Environment and Consumer Protection)
Soil ecosystems are exposed to a broad range of toxic compounds, among those various biocides. Building materials are vital source for soil contamination. About 25 % of the annual produced biocides are used in building materials. Biocides are utilized to protect the building materials against animal pests, algae, fungi and other microorganisms. Especially temporal and spatial input of biocidal building materials may heavily charge soils.
During the course of this project, representative test samples of building materials and component mixtures are incubated in a weathering chamber under different weather conditions and characterized in more detail using the high-throughput methods proteomics and microbiome analysis. In addition, highly sensitive biocide analysis is carried out using UPLC. In collaboration with the AG Noll, established and certified aquatic and terrestrial ecotoxicological tests are carried out. In cooperation with the German Environment Agency the used “omics”-methods are to be transferred into standard operating procedures.
Förster, Y., Schmidt, J. R., Wissenbach, D. K., Pfeiffer, S. E. M., Baumann, S., Hofbauer, L. C., Bergen, M. von, Kalkhof, S., and Rammelt, S., “Microdialysis Sampling from Wound Fluids Enables Quantitative Assessment of Cytokines, Proteins, and Metabolites Reveals Bone Defect-Specific Molecular Profiles,”PloS one, V. 11, No. 7, 2016, e0159580.
Hsu, M. J., Kratochvil, I., Winkler, S., Hempel, M., Kühne, H., Kalkhof, S., Baumann, S., Schubert, K., Bergen, M., and Christ, B., “Mesenchymale Stromalzellen begünstigen den Lipidabbau in isolierten Hepatozyten,”Z Gastroenterol, V. 56, No. 08, 2018, e252-e253.
Kalkhof, S., Schildbach, S., Blumert, C., Horn, F., Bergen, M. von, and Labudde, D., “PIPINO: A Software Package to Facilitate the Identification of Protein-Protein Interactions from Affinity Purification Mass Spectrometry Data,”BioMed research international, V. 2016, 2016, p. 2891918.
Kratochvil, I., Hofmann, T., Rother, S., Schlichting, R., Moretti, R., Scharnweber, D., Hintze, V., Escher, B. I., Meiler, J., Kalkhof, S., and Bergen, M. von, “Mono(2-ethylhexyl) phthalate (MEHP) and mono(2-ethyl-5-oxohexyl) phthalate (MEOHP) but not di(2-ethylhexyl) phthalate (DEHP) bind productively to the peroxisome proliferator-activated receptor γ,”Rapid communications in mass spectrometry : RCM, 2018.
May, J. C., Jurneczko, E., Stow, S. M., Kratochvil, I., Kalkhof, S., and McLean, J. A., “Conformational Landscapes of Ubiquitin, Cytochrome c, and Myoglobin: Uniform Field Ion Mobility Measurements in Helium and Nitrogen Drift Gas,”International journal of mass spectrometry, V. 427, 2018, pp. 79–90.
Rother, S., Samsonov, S. A., Hofmann, T., Blaszkiewicz, J., Köhling, S., Moeller, S., Schnabelrauch, M., Rademann, J., Kalkhof, S., Bergen, M. von, Pisabarro, M. T., Scharnweber, D., and Hintze, V., “Structural and functional insights into the interaction of sulfated glycosaminoglycans with tissue inhibitor of metalloproteinase-3 – A possible regulatory role on extracellular matrix homeostasis,”Acta biomaterialia, V. 45, 2016, pp. 143–154.
Schaarschmidt, J., Nagel, M. B. M., Huth, S., Jaeschke, H., Moretti, R., Hintze, V., Bergen, M. von, Kalkhof, S., Meiler, J., and Paschke, R., “Rearrangement of the Extracellular Domain/Extracellular Loop 1 Interface Is Critical for Thyrotropin Receptor Activation,”The Journal of biological chemistry, V. 291, No. 27, 2016, pp. 14095–14108.
Schmidt, J. R., Kliemt, S., Preissler, C., Moeller, S., Bergen, M. von, Hempel, U., and Kalkhof, S., “Osteoblast-released Matrix Vesicles, Regulation of Activity and Composition by Sulfated and Non-sulfated Glycosaminoglycans,”Molecular & cellular proteomics : MCP, V. 15, No. 2, 2016, pp. 558–572.
Wewering, F., Jouy, F., Wissenbach, D., Gebauer, S., Bergen, M. von, Luch, A., Kalkhof, S., and Zellmer, S., “A human hepatic in vitro co-culture system for the analysis of DILI related signaling,”Z Gastroenterol, V. 53, No. 12, 2015.
Wewering, F., Jouy, F., Caliskan, S., Kalkhof, S., Bergen, M. von, Luch, A., and Zellmer, S., “Hepatic co-cultures in vitro reveal suitable to detect Nrf2-mediated oxidative stress responses on the bladder carcinogen o-anisidine,”Toxicology in vitro : an international journal published in association with BIBRA, V. 40, 2017, pp. 153–160.
Wewering, F., Jouy, F., Wissenbach, D. K., Gebauer, S., Blüher, M., Gebhardt, R., Pirow, R., Bergen, M. von, Kalkhof, S., Luch, A., and Zellmer, S., “Characterization of chemical-induced sterile inflammation in vitro: application of the model compound ketoconazole in a human hepatic co-culture system,”Archives of toxicology, V. 91, No. 2, 2017, pp. 799–810.
Winkler, S., Kalkhof, S., Brückner, S., Hempel, M., Baumann, S., Bergen, M. von, and Christ, B., “Metabolic fingerprint of an immunodeficient NASH mouse model and impact after stem cell therapy,”Journal of Hepatology, V. 66, No. 1, 2017, S606-S607.
Wuchty, S., Müller, S. A., Caufield, J. H., Häuser, R., Aloy, P., Kalkhof, S., and Uetz, P., “Proteome Data Improves Protein Function Prediction in the Interactome of Helicobacter pylori,”Molecular & cellular proteomics : MCP, V. 17, No. 5, 2018, pp. 961–973.