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).
- Based on my application, should I use the NEBExpress® Cell-free E. coli Protein Synthesis System (NEB #E5360) or the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800)?
- What is the difference between NEBExpress® Cell-free E. coli Protein Synthesis System and the PURExpress In Vitro Protein Synthesis Kit?
- Protein Synthesis Reaction using PURExpress (E6800)
- Protein Synthesis Reaction using PURExpress (E3313)
- Protein Synthesis Reaction using PURExpress® ∆ (aa, tRNA) Kit (E6840)
- Protein Synthesis Reaction using PURExpress® ∆ RF123 Kit (E6850)
- Quick Start Protocol for NEBExpress® Cell-free E. coli Protein Synthesis System
- Use of the PURExpress® in vitro Protein Synthesis Kit, Disulfide Bond Enhancer and SHuffle® Competent E. coli for heterologous in vitro and in vivo cellulase expression.
- Using the PURExpress® In Vitro Protein Synthesis Kit for Heterologous In Vitro Expression and Functional Screening of FMN-dependent Oxidoreductase Variants
- NEBExpress® Cell-free E. coli Protein Synthesis System
- Accelerating DNA Construction to Protein Expression A Rapid 1-Day Workflow Using NEBridge Golden Gate Assembly
- Scaling down to scale up – Miniaturizing cell-free protein synthesis reactions with the Echo 525 Acoustic Liquid Handler
Avoid Common Obstacles in Protein Expression
Read how to avoid common obstacles in protein expression that prevent interactions with cellular machinery.
The Future of Cell-Free Protein Synthesis
Cell-free protein synthesis has the potential to become one of the most important high throughput technologies for functional genomics and proteomics.
- Protein Expression & Purification Brochure
- Purification Beads, Columns and Resins Brochure
- PURExpress® vs. NEBExpress® Application Chart
- Protein Expression and Purification Selection Chart
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This webinar discusses the fundamental workflows and advantages of cell-free (in vitro) protein synthesis versus traditional cellular expression methods.
Watch this tutorial explaining the streamlined workflow for our new NEBExpress® Cell-free Protein Synthesis System to learn how you can easily synthesize your protein in as little as 2 to 4 hours.
NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.