Cell-Free Technology

Synthelis Cell-free Protein Synthesis TechnologyCell-free expert

Cell-free protein expression systems (CFPS) exploit the cell transcription and translation machinery outside of the cells to express only one or several genes of interest in vitro. Thus, the protein expression machinery is only dedicated to the protein of interest. Cell-free technology also offers the advantages of an open system, contrary to the cell based systems where the cells constitute closed units.
This feature allows the possibility to incorporate elements specific to the target protein to increase the expression rate and obtain the proper folding. Cell-free expression systems have been the key to unlocking the production of a number of highly interesting molecules, due to its independence from physiological constraints; this broadens our horizons for the production of membrane proteins, or toxic proteins.

Synthelis’ innovative patented cell-free technology allows customized expression and characterization of active proteins, packaged in proteoliposomes  or delivered in a detergent solution.
This makes it particularly suitable for the expression of membrane proteins, which are well known to be challenging to produce in a usable form. The independence of cell-free technology from the physiological constraints of cell-based systems, also makes it generally relevant for any difficult-to-express proteins, including soluble, or toxic proteins.

Compared with other expression systems, Synthelis’ cell-free approach results in:

  • Fast expression of a broad range of protein variants
  • Fully preserved functionality, with full length proteins
  • Large protein amounts (10 to 100 mg) on a regular basis with a possible scaling up if requested
  • Rapid feasibility studies (up to 2-3 months)
  • Fast and economic protein labeling (for structural and functional purposes)
  • >90% success rate, depending on the type of protein.
  • Short production times for previously expressed proteins: 1-2 weeks per mg of protein*.

The following table shows what Synthelis can do with its cell-free system to express, or to modify, the protein you need for your project.

Required type of protein Key compounds used
Membrane proteins Detergents, lipids
Labeled proteins Modified amino acids
Modified proteins Modified amino acids
Unstable proteins Chaperones, protease inhibitors
Cytotoxic proteins  
Apo-proteins Cofactors
Disulfide bonded proteins Oxidative conditions, PDts

*1 mg of CXCR4 or Bak protein in one week for example. Times may vary depending on the expression rate of the protein.

 

References

Membrane protein production in Escherichia coli cell-free lysates Erik Henrich, Christopher Hein, Volker Dötsch, Frank Bernhard ,Volume 589, Issue 15, 8 July 2015, Pages 1713–1722

Cell-free protein synthesis: The state of the art James W Whittaker Biotechnol Lett. 2013 Feb;35(2):143-52. doi: 10.1007/s10529-012-1075-4. Epub 2012 Oct 21.