Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses IZI-BB
Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses IZI-BB

Dipl.-Ing. FH Doreen Wüstenhagen

leads the research group Eukaryotic Lysates

 

© Fraunhofer IZI-BB

Scientific focus and expertise:

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  • Knauer, J. F., Liers, C., Hahn, S., Wuestenhagen, D. A., Zemella, A., Kellner, H., Haueis, L., Hofrichter, M., & Kubick, S. (2022). Cell-free production of the bifunctional glycoside hydrolase GH78 from Xylaria polymorpha. Enzyme and Microbial Technology, 161, 110110. https://doi.org/10.1016/j.enzmictec.2022.110110
  • Krebs, S. K., Stech, M., Jorde, F., Rakotoarinoro, N., Ramm, F., Marinoff, S., Bahrke, S., Danielczyk, A., Wüstenhagen, D. A., & Kubick, S. (2022). Synthesis of an Anti-CD7 Recombinant Immunotoxin Based on PE24 in CHO and E. coli Cell-Free Systems. International Journal of Molecular Sciences, 23(22). https://doi.org/10.3390/ijms232213697
  • Dhandapani, P., Dondapati, S. K., Zemella, A., Bräuer, D., Wüstenhagen, D. A., Mergler, S., & Kubick, S. (2021). Targeted esterase-induced dye (TED) loading supports direct calcium imaging in eukaryotic cell-free systems. RSC Advances, 11(27), 16285–16296. https://doi.org/10.1039/d0ra08397f
  • Wüstenhagen, D. A., Lukas, P., Müller, C., Aubele, S. A., Hildebrandt, J.-P., & Kubick, S. (2020). Cell-free synthesis of the hirudin variant 1 of the blood-sucking leech Hirudo medicinalis. Scientific Reports, 10(1), 19818.  https://doi.org/10.1038/s41598-020-76715-w
  • Thoring, L., Wüstenhagen, D. A., Borowiak, M., Stech, M., Sonnabend, A., & Kubick, S. (2016). Cell-Free Systems Based on CHO Cell Lysates: Optimization Strategies, Synthesis of "Difficult-to-Express" Proteins and Future Perspectives. PloS One, 11(9), e0163670. dx.doi.org/10.1371/journal.pone.0163670 Artikel
  • Dondapati, S. K., Kreir, M., Quast, R. B., Wüstenhagen, D. A., Brüggemann, A., Fertig, N., & Kubick, S. (2014). Membrane assembly of the functional KcsA potassium channel in a vesicle-based eukaryotic cell-free translation system. Biosensors & Bioelectronics, 59, 174–183. https://doi.org/10.1016/j.bios.2014.03.004
  • Stech, M., Quast, R. B., Sachse, R., Schulze, C., Wüstenhagen, D. A., & Kubick, S. (2014). A continuous-exchange cell-free protein synthesis system based on extracts from cultured insect cells. PloS One, 9(5), e96635. https://doi.org/10.1371/journal.pone.0096635
  • Bechlars, S., Wüstenhagen, D. A., Drägert, K., Dieckmann, R., Strauch, E., & Kubick, S. (2013). Cell-free synthesis of functional thermostable direct hemolysins of Vibrio parahaemolyticus. Toxicon : Official Journal of the International Society on Toxinology, 76, 132–142. https://doi.org/10.1016/j.toxicon.2013.09.012
  • Brödel, A. K., Sonnabend, A., Roberts, L. O., Stech, M., Wüstenhagen, D. A., & Kubick, S. (2013). Ires-mediated translation of membrane proteins and glycoproteins in eukaryotic cell-free systems. PloS One, 8(12), e82234. https://doi.org/10.1371/journal.pone.0082234
  • Sachse, R., Wüstenhagen, D., Šamalíková, M., Gerrits, M., Bier, F. F., & Kubick, S. (2013). Synthesis of membrane proteins in eukaryotic cell‐free systems. Engineering in Life Sciences, 13(1), 39–48. https://doi.org/10.1002/elsc.201100235.
  • Zampatis, D. E., Rutz, C., Furkert, J., Schmidt, A., Wüstenhagen, D., Kubick, S., Tsopanoglou, N. E., & Schülein, R. (2012). The protease-activated receptor 1 possesses a functional and cleavable signal peptide which is necessary for receptor expression. FEBS Letters, 586(16), 2351–2359. https://doi.org/10.1016/j.febslet.2012.05.042
  • Brödel, A. K., Sonnabend, A., & Kubick, S. (2014). Cell-free protein expression based on extracts from CHO cells. Biotechnology and Bioengineering, 111(1), 25–36. https://doi.org/10.1002/bit.25013
  • Stech, M., Merk, H., Schenk, J. A., Stöcklein, W. F. M., Wüstenhagen, D. A., Micheel, B., Duschl, C., Bier, F. F., & Kubick, S. (2012). Production of functional antibody fragments in a vesicle-based eukaryotic cell-free translation system. Journal of Biotechnology, 164(2), 220–231. https://doi.org/10.1016/j.jbiotec.2012.08.020

 

  • Fermentation of eukaryotic cell lines in suspension culture
  • Production of translation-active lysates and their integration into cell-free systems
  • Development of eukaryotic cell-free translation systems
  • Testing of new cell lines for their in vitro expression capacity
  • Validation of DNA and mRNA templates
  • Integration of regulatory sequences, signal peptides, IRES sites, purification and fluorescence tags via generation of linear templates
  • Cell-free synthesis of difficult-to-express proteins, such as cytotoxic proteins and membrane proteins
  • Evaluation of protein synthesis using various cell-free systems (lysates from insect cells, CHO cells, cultured human cells; E. coli and wheat germ lysates) in batch and dialysis mode (CECF)
  • MS analyses of peptides, proteins (identification, characterization and investigation of modifications, such as glycosylation, phosporylation, palmitylation), protein-protein as well as protein-ligand interactions
  • Determination of synthesis yield by (14C)-protein labeling and TCA precipitation
  • Characterization of protein synthesis by gel electrophoresis, autoradiography and quantitative imaging in the phosphorimager Protein analysis by fluorescence microscopy and western blotting