Strategy exploration for developing robust lyophilized cell-free systems

Cell-free protein synthesis system is emerging as a powerful tool for rapid expression of therapeutic proteins. However, one drawback of cell-free technology is the necessity to store the major components below freezing in bulky aqueous solutions. To preserve the cell-free synthesis system for a longer time, the lyophilized cell-free system was developed, and its stability under freeze-drying conditions was explored. The novelty and the difference of this study from the others lie in determining more detailed storage conditions, which could provide a more accurate storage strategy. The effects of the treatment time, the storage temperature, and the component-mixing mode on the freeze-dried cell-free system were investigated. This study explored a robust storage strategy for freeze-dried cell-free systems and could further open up a better opportunity for on-demand synthesis of therapeutic proteins.