These extracellular signals must be transmitted across the leukocyte’s plasma membrane in a way that intracellular signaling cascades enable directional cell movement. Leukocytes are specialized in sensing a variety of guidance cues and to integrate environmental stimuli to navigate in a timely and spatially controlled manner. Leukocyte migration across vessels into and within peripheral and lymphoid tissues is essential for host defense against invading pathogens. 3Theodor Kocher Institute, University of Bern, Bern, Switzerland.2Faculty of Biology, University of Konstanz, Konstanz, Germany.1Biotechnology Institute Thurgau at the University of Konstanz, Kreuzlingen, Switzerland."The NMR solution structure of the artificial protein M7 matches the computationally designed model." Proteins.Guerric P. "A tetrapeptide fragment-based design method results in highly stable artificial proteins." Proteins 68(4): 839-49.ĥ Claudius Stordeur, Roman Dallüge, Olaf Birkenmeier, Hans Wienk, Rainer Rudolph, Christian Lange, Christian Lücke (2008). Rainer Rudolph, and Christian Lange (2007). "Affilin–Novel Binding Molecules Based on Human γ-B-Crystallin, an All β-Sheet Protein." Journal of Molecular Biology 372(1): 172-185.Ĥ Roman Dallüge, Jan Oschmann, Olaf Birkenmeier, Christian Lücke, Hauke Lilie, Stubbs, Carola Reimann, Gabriele Proetzel, Rainer Rudolph and Ulrike Fiedler (2007). "Engineering novel binding proteins from nonimmunoglobulin domains." Nature Biotechnology 23(10): 1257-1268.ģ Hilmar Ebersbach, Erik Fiedler, Tanja Scheuermann, Markus Fiedler, Milton T. Kaspar Binz, Patrick Amstutz & Andreas Plückthun (2005). "Artificial, non-antibody binding proteins for pharmaceutical and industrial applications." Trends in Biotechnology 23(10): 514-522.Ģ H. 4,5 Various library formats are developed for selection of artificial binding proteins.ġ Thomas Hey, Erik Fiedler, Rainer Rudolph and Markus Fiedler (2005). The scaffolds we evaluate and use currently are the human gamma-Crystallin 3 and the thermodynamically extremely stable computationally designed protein M7. With the aquired data we are able to optimise the selection procedure.Ĭhemical and genetical fusion methods are analyzed for advantages and disadvantages in ease of handling, avidity effects and other characteristics. Optimized automation protocols allow a high through-put in these steps and in return create large amounts of data on biochemical and biophysical properties of selected proteins.ĭifferent selection systems (ribosome display, various phage display methods) are analyzed for selection efficiency and characteristics of the selected binding proteins (affinity, solubility and thermodynamical stability). Selection and screening of libraries as well as production and purification of the so-called hit-variants is extremely suited for automation. Our work will also give us a deeper insight into molecular principles influencing protein-protein interactions as well as protein folding and stability. Which can then be used for the routine creation of artificial binding proteins. We aim to integrate optimized existing and novel innovative methods into a complete technology platform Applications for such selected artificial binding proteins can be found for example in therapy, diagnostics and research. The library is subsequently screened for binding activity against specific targets by evolutionary selection methods, e.g. This either generates a de novo binding patch or alters an existing patch. To achieve this we start from creating a protein variant library by randomizing the surface of the scaffold proteins. coliīy protein engineering and evolutionary selection techniques we are able to equip the scaffold proteins with the desired binding specificities. be easily producible in prokaryotes, e.g. be highly thermodynamically stable and free of disulfide bonds 1,2 Based on novel scaffold proteins they can circumvent some problems that antibodies are faced with today when applied in fields like medicine or biotechnology. InnoProfile Junior Research Group Artificial Binding ProteinsĪrtificial binding proteins are also referred to as the next-generation antibodies.
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