Preprint Review Version 2 Preserved in Portico This version is not peer-reviewed

The Evolution of Cell Free Biomanufacturing

Version 1 : Received: 31 March 2020 / Approved: 31 March 2020 / Online: 31 March 2020 (22:24:23 CEST)
Version 2 : Received: 29 April 2020 / Approved: 30 April 2020 / Online: 30 April 2020 (05:19:00 CEST)

A peer-reviewed article of this Preprint also exists.

Vilkhovoy, M.; Adhikari, A.; Vadhin, S.; Varner, J.D. The Evolution of Cell Free Biomanufacturing. Processes 2020, 8, 675. Vilkhovoy, M.; Adhikari, A.; Vadhin, S.; Varner, J.D. The Evolution of Cell Free Biomanufacturing. Processes 2020, 8, 675.


Cell free systems are a widely used research tool in systems and synthetic biology and a promising platform for manufacturing of proteins and chemicals. In the past, cell free biology was primarily used to better understand fundamental biochemical processes. Notably, E. coli cell free extracts were used in the 1960s to decipher the sequencing of the genetic code. Since then, the transcription and translation capabilities of cell free systems have been repeatedly optimized to improve energy efficiency and protein yield. Today, cell free systems, in combination with the rise of synthetic biology, have taken on a new role as a promising technology for just in time manufacturing of therapeutically important biologics and high-value small molecules. They have also been implemented in an industrial scale for the production of antibodies and cytokines. In this review, we discuss the evolution of cell free systems, advancements in cell free protein synthesis, and cell free metabolic engineering, and conclude with discussing the importance and feasibility of mathematical modeling in cell free systems.


cell free protein synthesis; cell free metabolic engineering; metabolic modeling


Engineering, Bioengineering

Comments (1)

Comment 1
Received: 30 April 2020
Commenter: Jeffrey Varner
Commenter's Conflict of Interests: Author
Comment: Summary of changes (reponse to reviewers)
1. We added a timeline figure (Fig. 1) to better illustrate the evolution of cell free technology development.
2. We added subject headings to aid the organization and readibility of the manuscript. 
3. We added text reviewing current outstanding issue in the field, for example scale-up of cell free reactions
4. We reorganized the mathematical modeling section
5. We reorgamized the conclusions section 
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