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Cell-free Membrane Protein Expression

Cell-free protein synthesis (CFPS) is a versatile technique derived from crude cell extracts and it has shown remarkable utility including the production of pharmaceuticals and protein libraries. As a complement to in vivo components, CFPS system offers some advanced benefits. Firstly, the reaction of CFPS is performed in an open environment, which allows some new components, such as NSAAs (non-standard amino acids), to be added or synthesized. Second, CFPS is free from constrains of cell-viability requirements, which allows protein synthesis to proceed with otherwise toxic reagents or protein products. Third, CFPS can use linear DNA fragments to perform a target gene expression, avoiding time-consuming gene cloning steps which are commonly required for in vivo protein synthesis. Finally, from a biomanufacturing perspective, CFPS separate catalyst synthesis (cell growth) from catalyst utilization (protein production). Some popular CFPS systems include E.coli (prokaryotic), wheat germ (WGE, eukaryotic), rabbit reticulocytes (RRL, eukaryotic), and insect cells (ICE, eukaryotic).

Cell-free synthesis of membrane proteins in presence of different lipid Figure 1: Cell-free synthesis of membrane proteins in presence of different lipid based membrane structures.

Creative Biostructure provides MemPro™ cell-free protein synthesis (CFPS) services for the manipulation and optimization of membrane proteins (MPs). Membrane proteins are a large group of major pharmaceutical targets due to their important functions in diverse biological processes (See the list of membrane proteins). A CFPS system allows for the direct monitoring of the protein synthesis process, and more importantly it provides an ideal environment for the efficient production of soluble and functional MPs by adding chemical additives to mimic a lipidic environment, using lipid-detergent systems, nanodiscs, liposomes and biological membranes. Our scientists will also help you choose the most appropriate cell-free system, taking into consideration the availability of materials, protein origin and complexity, downstream processing, etc.

Please see the list of membrane proteins provided by Creative Biostructure. Feel free to contact us for a detailed quote!

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Reference

  1. Sachse R, et al. (2014) “Membrane protein synthesis in cell-free systems: from bio-mimetic systems to bio-membranes”. FEBS Letters 588(17):2774-2781.
  2. Hong SH, et al. (2014) “Non-standard amino acid incorporation into proteins using Escherichia coli cell-free protein synthesis”. Front Chem 2:34.
  3. Carlson ED, et al. (2012) “Cell-free protein synthesis: applications come of age”. Biotechnol Adv 30(5):1185-1194. 

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