Solvent Tolerance of Freeze-Dried Cfps Reactions Enables Materials Applications (Industry Candidate)

Cell-free systems have growing importance as a way to power synthetic biology tools in the field. Lysates may be dried for storage, delivering biochemical activity for sensing or producing molecules on-demand upon rehydration at the point of need. Up to now, cell-free protein synthesis (CFPS) reactions have been studied as aqueous solutions in test tubes or absorbed onto paper or cloth. Embedding biological functionality into broadly-used materials, such as plastic polymers, has long been an attractive goal. Unfortunately, this goal has for the most part remained out of reach, often due to the fragility of biological systems outside of aqueous environments. In this work, we describe and utilize a surprising and useful feature of lyophilized cell-free lysate systems: tolerance to anhydrous organic solvents. To explore this newly discovered feature, a variety of solvents were tested and CFPS reaction components were screened for protein-protective properties. Tolerance to suspension in organic solvents may facilitate the use of polymers to deliver dry cell-free reactions in the form of coatings or fibers, among other processing possibilities. In pursuit of this application, preliminary casting into PLGA polymer is demonstrated as a proof of concept.