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A cell-free system is an in vitro technology widely used to study biological reactions that happen within cells while reducing the complex interactions found in a whole cell. Subcellular fractions can be isolated by ultracentrifugation to provide molecular machinery that can be used in reactions in the absence of many of the other cellular components.
Figure 1. Cell-free protein synthesis system for producing proteins or (poly)peptide-based materials. (Front. Chem., 2014)
Cell-free expression systems can be prepared by incubating a number of purified enzymes and coenzymes. Cell-free expression systems are developed as a novel cost-efficient biomanufacturing technology compared to cell-based fermentation used for thousands of years. Cell-free expression systems have several ADVANTAGES suitable in industrial applications:
Comparison of cell-free protein expression systems
|E. Coli cell lysate||Rabbit reticulocyte cell lysate||Hela cell lysate||CHO cell lysate|
|Protein modifications||None||Limited glycosylation||Glycosylation and phosphorylation||Glycosylation and phosphorylation|
|Recommended for high MW proteins||No||No||Yes||Yes|
|Produces functional proteins||Possible||Possible||Yes||Yes|
|Comments||Robust system||Flexible system||Higher protein yield per reaction||Most economocal cost per miligram produced|