Cell-Free Protein Expression

Cell-free protein synthesis (CFPS) is a protein expression approach that enables production of a target protein without the use of living cells. In CFPS, a solution containing all the cellular machinery needed to direct protein synthesis (e.g., ribosomes, tRNAs, enzymes, cofactors, amino acids, etc.) is used to transcribe and translate a supplied nucleic acid template (e.g., plasmid DNA, linear DNA or mRNA). In a simple biochemical reaction, CFPS generates desired proteins in just hours compared to traditional in vivo protein expression approaches that take several days or longer.

The NEBExpress^® Cell-free E. coli Protein Synthesis System (NEB #E5360) is a lysate-based system derived from E. coli cells that have been engineered for high in vitro expression performance. This system offers high expression levels, the ability to produce high molecular weight proteins, scalability, and is cost-effective for high throughput expression applications. Alternatively, CFPS can be achieved using purified protein synthesis machinery. The PURExpress^® In Vitro Protein Synthesis Kit (NEB #E6800) is a reconstituted mixture of numerous individually purified components that are required for E. coli transcription and translation. This system permits the production of proteins in the absence of detrimental E. coli activities (e.g., nucleases and proteases).