Abstract
Griffithsin is an algae-derived lectin with strong anti-viral activity against HIV and a positive safety profile. Multiple clinical studies are investigating griffithsin's utility as topical HIV microbicide. HIV microbicides are an extremely cost-sensitive market and plant-based griffithsin protein expression has the potential to meet those demands. The griffithsin product used in the clinic has been expressed and purified in N. benthamiana, using a TMV-based viral vector system, Geneware®. Outdoor pharming of biopharmaceuticals would further alleviate startup costs for biotechnology firms and may allow broader product accessibility. Therefore, this study assessed expression in a hybrid tobacco line, N. excelsiana, that is susceptible to TMV-based viral vectors and can be grown outdoors. In addition to using this hybrid line we expand on methods for in planta storage of griffithsin in leafy plants by ensiling kilogram quantities of griffithsin. The ensiling process allows year-round biomanufacturing, minimal environmental-controlled storage, and reduces the industry need for multiple growth areas to maintain multi-product manufacturing of plant-based pharmaceuticals. This study shows that griffithsin can be expressed in N. excelsiana and is stable, recoverable, and active from ensiled tissue. These studies can pave the way for future plant-based pharmaceuticals to be expressed and stored in this manner.
Highlights
Human Immunodeficiency Virus (HIV) is one of many diverse enveloped viruses that can cause life-threatening human disease
In the following study we explored the feasibility and practicality of silage as a method of storing GRFT expressed in N. excelsiana
N. excelsiana is an interspecific hybrid derived from a cross between N. excelsior and N. benthamiana
Summary
Human Immunodeficiency Virus (HIV) is one of many diverse enveloped viruses that can cause life-threatening human disease. Pre-exposure prophylaxis has been shown to be one of the most effective ways to prevent HIV transmission; various issues, such as cost, side effects, access, and adherence have led to less than ideal effectiveness of these drugs (Abdool Karim et al, 2010; Marrazzo et al, 2015; Vamvaka et al, 2016; Marrazzo, 2017). This is evident in rural and developing regions, Ensiling Nicotiana excelsiana Expressing Griffithsin where access to these drugs may be difficult or non-existent. There is a particular need to develop a highquality stable virucide that is able to be transported to diverse regions in less-than-ideal environmental conditions that would retain activity to combat the virus months or years after it has been manufactured
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