Roset Julià Laura, Grasemann Laura, Stellacci Francesco, Maerkl Sebastian J
Institute of Materials, School of Engineering, École Polytechnique Fédérale de Lausanne, Lausanne 1015, Switzerland.
Swiss National Center for Competence in Research (NCCR) Bio-Inspired Materials, University of Fribourg, Fribourg 1700, Switzerland.
ACS Synth Biol. 2025 Jan 17;14(1):290-295. doi: 10.1021/acssynbio.4c00618. Epub 2025 Jan 5.
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.
无细胞系统是合成生物学中功能强大的工具，具有广泛多样的应用。然而，这些系统，特别是无细胞PURE系统的一个重大瓶颈是其有限的反应寿命和产量。透析提供了一种延长反应寿命和提高产量的有前景的方法，但大多数定制透析系统需要使用3D打印机或微加工工具等精密设备。在本研究中，我们利用市售组件构建了一种易于组装的中型无细胞反应透析系统。通过采用定期更换进料溶液的透析方法，我们在PURE系统中实现了1.16 mg/mL绿色荧光蛋白（GFP）的蛋白质产量，并将蛋白质合成延长了至少连续12.5天，证明了该系统具有出色的稳定性。
