Cell-free protein synthesis (CFPS) is a platform biotechnology that enables a breadth of applications. However, field applications remain limited due to the poor shelf-stability of aqueous cell extracts required for CFPS. Lyophilization of E. coli extracts improves shelf life but remains insufficient for extended storage at room temperature. To address this limitation, we mapped the chemical space of ten low-cost additives with four distinct mechanisms of action in a combinatorial manner to identify formulations capable of stabilizing lyophilized cell extract. We report three key findings: (1) unique additive formulations that maintain full productivity of cell extracts stored at 4 °C and 23 °C; (2) additive formulations that enhance extract productivity by nearly 2-fold; (3) a machine learning algorithm that provides predictive capacity for the stabilizing effects of additive formulations that were not tested experimentally. These findings provide a simple and low-cost advance toward making CFPS field-ready and cost-competitive for biomanufacturing.

中文翻译：

---

通过延长保质期和提高大肠杆菌提取物的生产率来释放无细胞生物技术的应用。

无细胞蛋白质合成（CFPS）是一种平台生物技术，可实现广泛的应用。然而，由于CFPS所需的水性细胞提取物的储藏稳定性差，因此现场应用仍然受到限制。大肠杆菌提取物的冻干可延长保质期，但仍不足以在室温下长时间保存。为了解决这一局限性，我们以组合方式绘制了具有四种不同作用机理的十种低成本添加剂的化学空间，以鉴定能够稳定冻干细胞提取物的制剂。我们报告了三个主要发现：（1）独特的添加剂配方，可保持4°C和23°C下储存的细胞提取物的完整生产率；（2）可将提取物生产率提高近2倍的添加剂配方；（3）一种机器学习算法，可为未经实验测试的添加剂配方的稳定效果提供预测能力。这些发现为实现CFPS在生物制造领域的现场准备和具有成本竞争力提供了简单而低成本的进步。