Hägele Lorena, Pfleger Brian F, Takors Ralf
Institute of Biochemical Engineering, University of Stuttgart, 70569 Stuttgart, Germany.
Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
Bioengineering (Basel). 2024 Sep 3;11(9):892. doi: 10.3390/bioengineering11090892.
In recent years, the design-build-test-learn (DBTL) cycle has become a key concept in strain engineering. Modern biofoundries enable automated DBTL cycling using robotic devices. However, both highly automated facilities and semi-automated facilities encounter bottlenecks in clone selection and screening. While fully automated biofoundries can take advantage of expensive commercially available colony pickers, semi-automated facilities have to fall back on affordable alternatives. Therefore, our clone selection method is particularly well-suited for academic settings, requiring only the basic infrastructure of a biofoundry. The automated liquid clone selection (ALCS) method represents a straightforward approach for clone selection. Similar to sophisticated colony-picking robots, the ALCS approach aims to achieve high selectivity. Investigating the time analogue of five generations, the model-based set-up reached a selectivity of 98 ± 0.2% for correctly transformed cells. Moreover, the method is robust to variations in cell numbers at the start of ALCS. Beside , promising chassis organisms, such as and , were successfully applied. In all cases, ALCS enables the immediate use of the selected strains in follow-up applications. In essence, our ALCS approach provides a 'low-tech' method to be implemented in biofoundry settings without requiring additional devices.
近年来，设计-构建-测试-学习（DBTL）循环已成为菌株工程中的一个关键概念。现代生物铸造厂能够使用机器人设备实现自动化的DBTL循环。然而，高度自动化的设施和半自动化的设施在克隆选择和筛选方面都遇到了瓶颈。虽然全自动生物铸造厂可以利用昂贵的商用菌落挑选仪，但半自动化设施不得不依靠价格合理的替代方案。因此，我们的克隆选择方法特别适合学术环境，只需要生物铸造厂的基本基础设施。自动化液体克隆选择（ALCS）方法是一种直接的克隆选择方法。与复杂的菌落挑选机器人类似，ALCS方法旨在实现高选择性。通过研究五代的时间模拟，基于模型的设置对正确转化的细胞实现了98±0.2%的选择性。此外，该方法对ALCS开始时细胞数量的变化具有鲁棒性。此外，还成功应用了有前景的底盘生物，如 和 。在所有情况下，ALCS都能使所选菌株立即用于后续应用。本质上，我们的ALCS方法提供了一种“低技术”方法，可在生物铸造厂环境中实施，无需额外设备。
