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(599ax) Lyophilized Cell-Free Systems for Robust, Modular Protein Expression

Authors: 
Smith, M. T., Brigham Young University
Bundy, B. C., Brigham Young University
Werner, C., Brigham Young University

Cell-free Protein Synthesis (CFPS) is a modular and scalable transcription/translation system with many propitious features for protein expression and engineering. CFPS reactions are not restricted by a cellular membrane and thereby mitigate or avoid transmembrane transport limitations and cytotoxicity concerns. The “open” nature of this system provides direct control over reagents, which can be prepared as individual modular elements (eg – cell-extract, energy source, genetic material, etc). Reaction volumes have been demonstrated to scale from microliters to 100 liters. One restriction of traditional CFPS systems is the need to store the modular elements below freezing. We hypothesized that lyophilizing (ie – freeze drying) modular CFPS elements would create a robust CFPS system that could be stored long-term above freezing. Here, we demonstrate a straight-forward a lyophilized CFPS system. A straightforward lyophilization technique allows for robust modular elements that retain protein synthesis viability when stored above freezing. Further benefits of lyophilized CFPS include significantly reduced storage volume and sterilization of our cell-extracts. This technology has the potential to facilitate robust portable protein expression devices such as pharmacy-on-a-chip and personalized point-of-care therapeutic expression.