scholarly journals Cell-free protein synthesis: advances on production process for biopharmaceuticals and immunobiological products

BioTechniques ◽  
2021 ◽  
Vol 70 (2) ◽  
pp. 126-133
Author(s):  
Camila Hiromi Chiba ◽  
Marcos Camargo Knirsch ◽  
Adriano Rodrigues Azzoni ◽  
Antonio R Moreira ◽  
Marco Antonio Stephano

Biopharmaceutical products are of great importance in the treatment or prevention of many diseases and represent a growing share of the global pharmaceutical market. The usual technology for protein synthesis (cell-based expression) faces certain obstacles, especially with ‘difficult-to-express’ proteins. Cell-free protein synthesis (CFPS) can overcome the main bottlenecks of cell-based expression. This review aims to present recent advances in the production process of biologic products by CFPS. First, key aspects of CFPS systems are summarized. A description of several biologic products that have been successfully produced using the CFPS system is provided. Finally, the CFPS system's ability to scale up and scale down, its main limitations and its application for biologics production are discussed.

1996 ◽  
Vol 799 (1 Enzyme Engine) ◽  
pp. 406-412
Author(s):  
HIDEO NAKANO ◽  
YASUAKI KAWARASAKI ◽  
TSUNEO YAMANE

RSC Advances ◽  
2020 ◽  
Vol 10 (66) ◽  
pp. 40588-40596
Author(s):  
Tony Köhler ◽  
Thomas Heida ◽  
Sandra Hoefgen ◽  
Niclas Weigel ◽  
Vito Valiante ◽  
...  

We describe a bottom-up approach towards functional enzymes utilizing microgels as carriers for genetic information that enable cell-free protein synthesis, in situ immobilization, and utilization of functional deGFP-MatB.


AIChE Journal ◽  
2021 ◽  
Author(s):  
Carlos A. Duran‐Villalobos ◽  
Olotu Ogonah ◽  
Beatrice Melinek ◽  
Daniel G. Bracewell ◽  
Trevor Hallam ◽  
...  

2021 ◽  
Author(s):  
Takeshi Tabuchi ◽  
Yohei Yokobayashi

Synthetic riboswitches can be used as chemical gene switches in cell-free protein synthesis systems. We provide a current perspective on the state of cell-free riboswitch technologies and their future directions.


2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Liyuan Zhang ◽  
Xiaomei Lin ◽  
Ting Wang ◽  
Wei Guo ◽  
Yuan Lu

AbstractCell-free protein synthesis (CFPS) systems have become an ideal choice for pathway prototyping, protein production, and biosensing, due to their high controllability, tolerance, stability, and ability to produce proteins in a short time. At present, the widely used CFPS systems are mainly based on Escherichia coli strain. Bacillus subtilis, Corynebacterium glutamate, and Vibrio natriegens are potential chassis cells for many biotechnological applications with their respective characteristics. Therefore, to expand the platform of the CFPS systems and options for protein production, four prokaryotes, E. coli, B. subtilis, C. glutamate, and V. natriegens were selected as host organisms to construct the CFPS systems and be compared. Moreover, the process parameters of the CFPS system were optimized, including the codon usage, plasmid synthesis competent cell selection, plasmid concentration, ribosomal binding site (RBS), and CFPS system reagent components. By optimizing and comparing the main influencing factors of different CFPS systems, the systems can be optimized directly for the most influential factors to further improve the protein yield of the systems. In addition, to demonstrate the applicability of the CFPS systems, it was proved that the four CFPS systems all had the potential to produce therapeutic proteins, and they could produce the receptor-binding domain (RBD) protein of SARS-CoV-2 with functional activity. They not only could expand the potential options for in vitro protein production, but also could increase the application range of the system by expanding the cell-free protein synthesis platform.


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