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research article

Long-Term Protein Synthesis with PURE in a Mesoscale Dialysis System

Roset Julià, Laura  
•
Grasemann, Laura  
•
Stellacci, Francesco  
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January 17, 2025
ACS Synthetic Biology

Cell-free systems are powerful tools in synthetic biology with versatile and wide-ranging applications. However, a significant bottleneck for these systems, particularly the PURE cell-free system, is their limited reaction lifespan and yield. Dialysis offers a promising approach to prolong reaction lifetimes and increase yields, yet most custom dialysis systems require access to sophisticated equipment like 3D printers or microfabrication tools. In this study, we utilized an easy-to-assemble, medium-scale dialysis system for cell-free reactions using commercially available components. By employing dialysis with periodic exchange of the feeding solution, we achieved a protein yield of 1.16 mg/mL GFP in the PURE system and extended protein synthesis for at least 12.5 consecutive days, demonstrating the system’s excellent stability.

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Type
research article
DOI
10.1021/acssynbio.4c00618
Scopus ID

2-s2.0-85215070645

PubMed ID

39757539

Author(s)
Roset Julià, Laura  

École Polytechnique Fédérale de Lausanne

Grasemann, Laura  

École Polytechnique Fédérale de Lausanne

Stellacci, Francesco  

École Polytechnique Fédérale de Lausanne

Maerkl, Sebastian J.  

École Polytechnique Fédérale de Lausanne

Date Issued

2025-01-17

Published in
ACS Synthetic Biology
Volume

14

Issue

1

Start page

290

End page

295

Subjects

cell-free expression

•

continuous dialysis

•

extended expression

•

lysate

•

PURE

•

scale-up

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
SUNMIL  
LBNC  
Available on Infoscience
January 25, 2025
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/244392
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