Heat-stable preservation of protein expression systems for portable therapeutics production

Many biotechnology capabilities are limited by stringent storage needs of reagents, largely prohibiting use outside of specialized laboratories. Focusing on a large class of protein-based biotechnology applications, we address this issue by developing a method for preserving cell-free protein expression systems under months of heat stress. Our approach realizes an unprecedented degree of long term heat stability by leveraging the sugar alcohol trehalose, a simple, low-cost, open-air drying step, and strategic separation of sets of reaction components during drying. The resulting preservation capacity opens the door for efficient production of a wide range of on-demand proteins under adverse conditions, for instance during emergency outbreaks or in remote or otherwise inaccessible locations. As such, our preservation method stands to advance a great number of different cell-free technologies, including remediation efforts, point of care therapeutics, and large-scale biosensing. To demonstrate this application potential, we use cell-free reagents subjected to months of heat stress and atmospheric conditions to produce sufficient concentrations of a pyocin protein to kill Pseudomonas aeruginosa , one of the most troublesome pathogens for traumatic and burn wound injuries. Our work makes possible new biotechnology applications that demand both ruggedness and scalability.