These are exiting times for humankind, after the recent revolution of molecular biology current research in synthetic biology aims at developing new control elements and new modules to existing genomes or build up wholly new genomes. DNA ligation via click-chemistry, the expansion of the genetic alphabet, synthetic production of whole genomes, and the use of unnatural amino acids, are a few very exciting examples of the new frontiers in biology.
Within the field of synthetic biology new methods are arising to translate and transcribe (new) DNA to usable protein. The synthetic biology workflow has its own language and the product of many processes are most often proteins. Protein production in more controlled processes has been a push to develop engineered expression systems. In addition Cell Free Protein Synthesis (or CFPS) is emerging as a promising and flexible technology for protein synthesis at different scales (micro to manufacturing). CFPS allows in vitro optimization of different cell extracts towards the exclusive production of a single protein. The CFPS bypass cell walls, organelle structures, and avoids the issues of cellular genetic regulation. The absence of cellular barriers allows easy incorporation of substrates, efficient product removal, rapid system monitoring and convenient sampling.
These efforts have resulted in in vitro synthesis of complex (prokaryotic and eukaryotic) proteins containing disulphide bridges, N-glycans, unnatural amino acids, and different types of radioactive/chemical labels. Compared to heterologous host systems, the relatively high protein yields and 10 fold faster gene-to-product rate in CFPS also make it a powerful method for small-volume and large sample number screening methods thereby speeding protein evolution. In addition, CFPS has also opened the possibility of new commercial production platforms (up to 100 L scale) for protein therapeutics and vaccines.
One of the immediate challenges in the field of the production of pharmaceutical proteins is the development of genomically re-coded organisms for the preparation of highly active extracts to be used in CFPS. It is my opinion, and focus of ongoing research, that the open field of the development of engineered strains for the production of lysates for current- and novel-CFPS application needs immediate exploration.
(c) Marco G. Casteleijn. 2014-2017
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