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Accelerating the Production of Druggable Targets: Eukaryotic Cell-Free Systems Come into Focus

Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI-BB), Am Mühlenberg 13, D-14476 Potsdam, Germany
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Methods Protoc. 2019, 2(2), 30; https://doi.org/10.3390/mps2020030
Received: 15 February 2019 / Revised: 5 April 2019 / Accepted: 10 April 2019 / Published: 16 April 2019
(This article belongs to the Special Issue Cell-Free Synthetic Biology)
In the biopharmaceutical pipeline, protein expression systems are of high importance not only for the production of biotherapeutics but also for the discovery of novel drugs. The vast majority of drug targets are proteins, which need to be characterized and validated prior to the screening of potential hit components and molecules. A broad range of protein expression systems is currently available, mostly based on cellular organisms of prokaryotic and eukaryotic origin. Prokaryotic cell-free systems are often the system of choice for drug target protein production due to the simple generation of expression hosts and low cost of preparation. Limitations in the production of complex mammalian proteins appear due to inefficient protein folding and posttranslational modifications. Alternative protein production systems, so-called eukaryotic cell-free protein synthesis systems based on eukaryotic cell-lysates, close the gap between a fast protein generation system and a high quality of complex mammalian proteins. In this study, we show the production of druggable target proteins in eukaryotic cell-free systems. Functional characterization studies demonstrate the bioactivity of the proteins and underline the potential for eukaryotic cell-free systems to significantly improve drug development pipelines. View Full-Text
Keywords: in vitro translation; protein production; drug development; cell-free protein synthesis; eukaryotic lysates; microsomes; growth factors; enzymes in vitro translation; protein production; drug development; cell-free protein synthesis; eukaryotic lysates; microsomes; growth factors; enzymes
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Thoring, L.; Zemella, A.; Wüstenhagen, D.; Kubick, S. Accelerating the Production of Druggable Targets: Eukaryotic Cell-Free Systems Come into Focus. Methods Protoc. 2019, 2, 30.

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