Thornton Ella Lucille, Paterson Sarah Maria, Gidden Zoe, Horrocks Mathew H, Laohakunakorn Nadanai, Regan Lynne
Centre for Synthetic and Systems Biology, Institute of Quantitative Biology, Biochemistry and Biotechnology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
School of Chemistry, University of Edinburgh, Edinburgh, United Kingdom.
Front Bioeng Biotechnol. 2022 Jul 7;10:915035. doi: 10.3389/fbioe.2022.915035. eCollection 2022.
We present a new method for the surface capture of proteins in cell-free protein synthesis (CFPS). We demonstrate the spontaneous self-assembly of the protein BslA into functionalizable surfaces on the surface of a CFPS reaction chamber. We show that proteins can be covalently captured by such surfaces, using "Catcher/Tag" technology. Importantly, proteins of interest can be captured either when synthesised by CFPS above the BslA surfaces, or when added as pure protein. The simplicity and cost efficiency of this method suggest that it will find many applications in cell-free-based methods.
我们提出了一种用于无细胞蛋白质合成(CFPS)中蛋白质表面捕获的新方法。我们展示了蛋白质BslA在CFPS反应室表面自发自组装成可功能化表面。我们表明,使用“捕获器/标签”技术,蛋白质可以通过此类表面共价捕获。重要的是,感兴趣的蛋白质既可以在BslA表面上方通过CFPS合成时捕获,也可以作为纯蛋白质添加时捕获。该方法的简单性和成本效益表明,它将在基于无细胞的方法中找到许多应用。