The Potential of Eukaryotic Cell-Free Systems as a Rapid Response to Novel Zoonotic Pathogens: Analysis of SARS-CoV-2 Viral Proteins

Abstract

The ongoing pandemic caused by the novel coronavirus (SARS-CoV-2) has led to more than 445 million infections and the underlying disease, COVID-19, resulted in more than 6 million deaths worldwide. The scientific world is already predicting future zoonotic diseases. Hence, rapid response systems are needed to tackle future epidemics and pandemics. Here, we present the use of eukaryotic cell-free systems for the rapid response to novel zoonotic diseases represented by SARS-CoV-2. Non-structural, structural and accessory proteins encoded by SARS-CoV-2 were synthesized by cell-free protein synthesis in a fast and efficient manner. The inhibitory effect of the non-structural protein 1 on protein synthesis could be shown in vitro. Structural proteins were quantitatively detected by commercial antibodies, therefore facilitating cell-free systems for the validation of available antibodies. The cytotoxic envelope protein was characterized in electrophysiological planar lipid bilayer measurements. Hence, our study demonstrates the potential of eukaryotic cell-free systems as a rapid response mechanism for the synthesis, functional characterization and antibody validation against a viral pathogen.

Scaling eukaryotic cell‐free protein synthesis achieved with the versatile and high‐yielding tobacco BY‐2 cell lysate

“…This circumvents confounding variables like transfection efficiencies, indicating a more flexible screening platform than cell‐based methods and with particular relevance to the monoclonal antibody discovery sector, especially as our CB2 work shows that function can be assess in‐lysate without purification. Streamlined protein variant expression also has relevance to pandemic preparedness for example, rapid production of SARS‐CoV‐2 RBD mutants from emerging viral strains for diagnostic or therapeutic purposes (Ramm et al, 2022). The use of CFPS has even been posited for individualized vaccines and distributed medicines manufacturing, presenting new medical paradigms if regulation can be safely adapted to meet this potential (Kanter et al, 2007; Ogonah et al, 2017; Zawada et al, 2022).…”

“…Streamlined protein variant expression also has relevance to pandemic preparedness for example, rapid production of SARS-CoV-2 RBD mutants from emerging viral strains for diagnostic or therapeutic purposes (Ramm et al, 2022). The use of CFPS has even been posited for individualized vaccines and distributed medicines manufacturing, presenting new medical paradigms if regulation can be safely adapted to meet this potential (Kanter et al, 2007;Ogonah et al, 2017;Zawada et al, 2022).…”

ALiCE®: A versatile, high yielding and scalable eukaryotic cell-free protein synthesis (CFPS) system

“…Streamlined protein variant expression also has relevance to pandemic preparedness e.g. rapid production of SARS-CoV-2 RBD mutants from emerging viral strains for diagnostic or therapeutic purposes 36 . The use of CFPS has even been posited for individualised vaccines and distributed medicines manufacturing, presenting new medical paradigms if regulation can be safely adapted to meet this potential [37][38][39] .…”