AUTHOR=Thornton Ella Lucille , Paterson Sarah Maria , Gidden Zoe , Horrocks Mathew H. , Laohakunakorn Nadanai , Regan Lynne TITLE=Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins JOURNAL=Frontiers in Bioengineering and Biotechnology VOLUME=10 YEAR=2022 URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2022.915035 DOI=10.3389/fbioe.2022.915035 ISSN=2296-4185 ABSTRACT=

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 in situ 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.