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

Abstract

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.