Lyophilized Escherichia coli-based cell-free systems for robust, high-density, long-term storage.
Cell-free protein synthesis (CFPS) is a versatile tool for rapid recombinant protein production and engineering. One drawback of cell-free technology is the necessity to store the major components-cell extracts and energy systems-below freezing in bulky aqueous solutions. Here we describe simple methods for lyophilizing extracts and preparing powdered energy systems for CFPS. These techniques allow for high-density storage of cell-free systems that are more robust against temperature and bacterial degradation. Our methods have the potential to decrease storage expenses, allow for longer shelf-life of cell extracts at room temperature, and enable durable portable protein production technologies.
Department of Chemical Engineering, Brigham Young University, Provo, Utah.
Mark Thomas
MT
Smith
Department of Chemical Engineering, Brigham Young University, Provo, Utah.
Scott D
SD
Berkheimer
Department of Chemical Engineering, Brigham Young University, Provo, Utah.
Christopher J
CJ
Werner
Department of Chemical Engineering, Brigham Young University, Provo, Utah.
Bradley C
BC
Bundy
D001709:Biotechnology; D002457:Cell Extracts; D002474:Cell-Free System / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D005612:Freeze Drying / Q000379:methods*; D011994:Recombinant Proteins; D013696:Temperature
D002457:Cell Extracts; D011994:Recombinant Proteins
GFP reporter protein; cell-extract preparation; cell-free energy system; cell-free protein synthesis; in vitro transcription / translation; lyophilization