(319a) Producing Protein Therapeutics without Cells

Recent engineering advances in in vitro or "cell-free" protein synthesis systems are enabling new possibilities for protein therapeutic production. Here we describe the development of lyophilized cell-free systems which provide the ability to economically stock-pile reagents for rapid response against an emerging threat and also enable facile transportation for just-add-water therapeutic production potentially anywhere. Importantly, the lyophilization processes also sterilizes the reagents which enables the use of genetically modified organisms to create cell-free reagents but prevents the release of genetically modified organisms to the environment during transit or storage. Additionally, a cell-free protein synthesis approach enables the production of complex, difficult-to-fold, and cytotoxic proteins at high production yields, solubilities, and activities. A case study producing Onconase (a promising cancer and anogenital wart biotherapeutic) using a lyophilized cell-free system is presented.