Cell-free protein synthesis: Applications come of age

Abstract

Cell-free protein synthesis has emerged as a powerful technology platform to help satisfy the growing demand for simple and efficient protein production. While used for decades as a foundational research tool for understanding transcription and translation, recent advances have made possible cost-effective microscale to manufacturing scale synthesis of complex proteins. Protein yields exceed grams protein produced per liter reaction volume, batch reactions last for multiple hours, costs have been reduced orders of magnitude, and reaction scale has reached the 100-liter milestone. These advances have inspired new applications in the synthesis of protein libraries for functional genomics and structural biology, the production of personalized medicines, and the expression of virus-like particles, among others. In the coming years, cell-free protein synthesis promises new industrial processes where short protein production timelines are crucial as well as innovative approaches to a wide range of applications.

Emerging technology of in situ cell free expression protein microarrays

“…As expected, not all the expressed proteins by cell free expression system are properly folded which includes oxidative folding pathways to allow for isomerization and formation of disulfide bonds. Considerable progress has been made towards enhancing the folding of eukaryotic proteins with multiple disulfide bonds (Carlson et al, 2011). In addition to this, new advances have been made to form and isomerize disulfide bonds by using natural enzymes (e.g.…”

“…This will have a positive outcome for the generation of new vaccines in the future. Another major application is the selection of better therapeutic antibodies from libraries of single chain antibodies (Carlson et al, 2011). Antibody profiling is another major area where these technologies can make a real difference.…”

Construction de cellules synthétiques

“…Le développement de systèmes d'expression encore plus puissants est nécessaire à la construction de cellules minimales. De tels travaux sont bénéfiques pour de nouvelles applications in vitro comme la production de molécules thérapeutiques telles que des vaccins [49,50]. L'une des clés du développement de ce type de recherche réside dans la création de nouvelles voies métaboliques afin de stimuler et accroître l'expression in vitro.…”

Protein-Based Textiles: Bio-Inspired and Bio-Derived Materials for Medical and Non-Medical Applications

“…[17][18][19] Synthesized proteins can be processed using traditional manufacturing techniques, such as wet spinning or electrospinning, to produce protein based fibers for medical applications, such as vascular grafts or tissue engineered scaffolds. [20][21][22] In this review, we will examine the design rules mediating protein self-assembly on the molecular level, and the current technologies available to build protein-based textiles, and the tools available to test these properties across multiple spatial scales.…”