The past 30 years have seen the growth of plant molecular farming as an approach to the production of recombinant proteins for pharmaceutical and biotechnological uses. Much of this effort has focused on producing vaccine candidates against viral diseases, including those caused by enveloped viruses. These represent a particular challenge given the difficulties associated with expressing and purifying membrane-bound proteins and achieving correct assembly. Despite this, there have been notable successes both from a biochemical and a clinical perspective, with a number of clinical trials showing great promise. This review will explore the history and current status of plant-produced vaccine candidates against enveloped viruses to date, with a particular focus on virus-like particles (VLPs), which mimic authentic virus structures but do not contain infectious genetic material.
Summary
We have investigated the use of transient expression to produce virus‐like particles (VLPs) of severe acute respiratory syndrome coronavirus 2, the causative agent of COVID‐19, in
Nicotiana benthamiana
. Expression of a native form of the spike (S) protein, either alone or in combination with the envelope (E) and membrane (M) proteins, all of which were directed to the plant membranes via their native sequences, was assessed. The full‐length S protein, together with degradation products, could be detected in total protein extracts from infiltrated leaves in both cases. Particles with a characteristic ‘crown‐shaped’ or ‘spiky’ structure could be purified by density gradient centrifugation. Enzyme‐linked immunosorbent assays using anti‐S antibodies showed that threefold higher levels of VLPs containing the full‐length S protein were obtained by infiltration with S alone, compared to co‐infiltration of S with M and E. The S protein within the VLPs could be cleaved by furin
in vitro
and the particles showed reactivity with serum from recovering COVID‐19 patients, but not with human serum taken before the pandemic. These studies show that the native S protein expressed in plants has biological properties similar to those of the parent virus. We show that the approach undertaken is suitable for the production of VLPs from emerging strains and we anticipate that the material will be suitable for functional studies of the S protein, including the assessment of the effects of specific mutations. As the plant‐made material is noninfectious, it does not have to be handled under conditions of high containment.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.