Cell-free systems are complex; hence the increased uptake in recent years of cell-free kits
The wide availability and convenience of cell-free kits has led to their increased use over the last few years. However, despite the multitude of cell-free kits on the market, they do not allow as much flexibility as custom solutions. Indeed, the base reaction buffer...
Cell-free systems do not produce functional protein complexes
Cell-free systems not only enable the production of proteins, but also protein complexes composed of around 2 to 1500 proteins. Indeed, the open nature of cell-free systems is advantageous for the co-expression of proteins since the ratios of DNA templates and the...
Cell-free systems only work with plasmids
It is true that cell-free protein synthesis works with genetic materials such as plasmid DNA but also with PCR-amplified linear DNA sequences. However, when cell-free expression assays started with PCR fragments, the expression yields were much lower than from plasmid...
Cell-free systems are only relevant for membrane proteins
Cell-free protein synthesis (CFPS) methodology offers the possibility to produce proteins that would be difficult or even impossible to synthesize in cell-based systems. These so-called “difficult-to-express proteins” include membrane proteins, but also cytotoxic...
Cell-free systems deliver low expression yields
It is generally considered that cell-free protein synthesis systems are capable of generating only a small expression yield. Even if this was true in the 2000s, it is certainly no longer the case today (Carlson et al., 2012) as we will explain in the following paper....
Cell-free systems only use bacterial extract.
Escherichia coli (E. Coli) is the most popular cell extract source for cell-free expression system. However, extracts from wheat germ, rabbit reticulocytes or insect cells are also commonly used. Other mammalian cell lines are also more recently used for cell-free...
Cell-free systems are not compatible with Good Manufacturing Practices (GMP grade)
In the last decade, major advances have been made in the field of cell-free protein synthesis. Although the manufacturing of therapeutic proteins has been dominated for many years by cell-based systems, mainly using mammalian and bacterial cell-type systems, cell-free...
Cell-free systems are not suitable for large-scale industrial applications
Although traditional cell-free protein synthesis is only considered appropriate for small scale protein production, technological breakthroughs made since the turn of the century have drastically accelerated the development of this production platform, making it now...