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

作者：Laura Roset Julià, Laura Grasemann, Francesco Stellacci, Sebastian J. Maerkl · 发表于：ACS Synthetic Biology · 年份：2025 · DOI：10.1021/acssynbio.4c00618 · 被引用次数：6 · 研究领域：Protein Structure and Dynamics、Enzyme Structure and Function、Biochemical and Molecular Research

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.