TY - JOUR
T1 - Self-Assembling Protein Surfaces for In Situ Capture of Cell-Free-Synthesized Proteins
AU - Thornton, Ella Lucille
AU - Paterson, Sarah Maria
AU - Gidden, Zoe
AU - Horrocks, Mathew H.
AU - Laohakunakorn, Nadanai
AU - Regan, Lynne
N1 - Funding Information:
ET acknowledges the support of the DSTL during her PhD. ZG acknowledges support of the BBSRC via the EastBio DTP (BB/M010996/1). We also acknowledge the School of Biological Sciences and The Edinburgh Protein Production Facility at the University of Edinburgh. LR, ET, SP, and NL acknowledge support of the Leverhulme Trust (RPG-2021-230). NL is supported by a UKRI Future Leaders Fellowship (MR/V027107/1). MH acknowledges Jim Love and UCB Biopharma for funding the microscope used in the FRAP experiments.
Funding Information:
ET acknowledges the support of the DSTL during her PhD. ZG acknowledges support of the BBSRC via the EastBio DTP (BB/M010996/1). We also acknowledge the School of Biological Sciences and The Edinburgh Protein Production Facility at the University of Edinburgh. LR, ET, SP, and NL acknowledge support of the Leverhulme Trust (RPG-2021-230). NL is supported by a UKRI Future Leaders Fellowship (MR/V027107/1). MH acknowledges Jim Love and UCB Biopharma for funding the microscope used in the FRAP experiments.
Publisher Copyright:
Copyright © 2022 Thornton, Paterson, Gidden, Horrocks, Laohakunakorn and Regan.
PY - 2022/7/7
Y1 - 2022/7/7
N2 - 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.
AB - 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.
U2 - 10.3389/fbioe.2022.915035
DO - 10.3389/fbioe.2022.915035
M3 - Article
SN - 2296-4185
VL - 10
JO - Frontiers in Bioengineering and Biotechnology
JF - Frontiers in Bioengineering and Biotechnology
ER -