Tools for tomorrow: expression without cells

Cell & Gene Therapy Insights 2020; 6(11), 1623–1633


Published: 8 January 2021
Jade Tuck, Stuart Jamieson, Philip Probert

Conventional biologics manufacturing approaches exploit the transcriptional and translational machinery of viable cells as the basis of production, be that in prokaryotic or eukaryotic hosts. Whilst these approaches are industry standard and decades of experience have led to the realisation of high yields, the requirement of growing cells brings with it variability and unpredictability, longer time frames for development and production, and physiological ranges in which processes must be run, but which may not necessarily provide optimal conditions for biomolecule expression or folding. Furthermore, their nature renders production of certain types of product, particularly RNA based therapeutics challenging as a consequence of the endogenous degradative pathways in cell systems. These factors, particularly within the context of gene therapies, don’t preclude development, but hinder identification, development and commercialisation of the next generation of advanced therapies.

Cell free synthesis systems, be they for the production of nucleic acids or proteins, have been used in academic research for decades, as tools initially to understand the fundamental nature of transcription and translation and subsequently for the production of small quantities of both nucleic acids and various proteins. With the development of these approaches and changing needs of biologic drug identification and manufacturing process development, cell free approaches are becoming more relevant and this is particularly true as much for conventional biologics as well as advanced cell and gene therapies. In this article we provide a brief introduction to the aforementioned technologies with particular focus on in-vitro transcription (IVT) and cell-free protein synthesis (CFPS) and their current and future application to supporting the identification and manufacture of advanced therapeutics.