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Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins

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 surfa...

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Detalles Bibliográficos
Autores principales: Thornton, Ella Lucille, Paterson, Sarah Maria, Gidden, Zoe, Horrocks, Mathew H., Laohakunakorn, Nadanai, Regan, Lynne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9300835/
https://www.ncbi.nlm.nih.gov/pubmed/35875503
http://dx.doi.org/10.3389/fbioe.2022.915035
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author Thornton, Ella Lucille
Paterson, Sarah Maria
Gidden, Zoe
Horrocks, Mathew H.
Laohakunakorn, Nadanai
Regan, Lynne
author_facet Thornton, Ella Lucille
Paterson, Sarah Maria
Gidden, Zoe
Horrocks, Mathew H.
Laohakunakorn, Nadanai
Regan, Lynne
author_sort Thornton, Ella Lucille
collection PubMed
description 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.
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spelling pubmed-93008352022-07-22 Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins Thornton, Ella Lucille Paterson, Sarah Maria Gidden, Zoe Horrocks, Mathew H. Laohakunakorn, Nadanai Regan, Lynne Front Bioeng Biotechnol Bioengineering and Biotechnology 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. Frontiers Media S.A. 2022-07-07 /pmc/articles/PMC9300835/ /pubmed/35875503 http://dx.doi.org/10.3389/fbioe.2022.915035 Text en Copyright © 2022 Thornton, Paterson, Gidden, Horrocks, Laohakunakorn and Regan. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Thornton, Ella Lucille
Paterson, Sarah Maria
Gidden, Zoe
Horrocks, Mathew H.
Laohakunakorn, Nadanai
Regan, Lynne
Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title_full Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title_fullStr Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title_full_unstemmed Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title_short Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
title_sort self-assembling protein surfaces for in situ capture of cell-free-synthesized proteins
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9300835/
https://www.ncbi.nlm.nih.gov/pubmed/35875503
http://dx.doi.org/10.3389/fbioe.2022.915035
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