Environmental factors controlling the production of recombinant proteins in Nicotiana benthamiana plants grown hydroponically in mobile gullies

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The overall goal of this project is to determine the main factors regulating transient expression and accumulation of recombinant vaccine antigens and other biopharmaceuticals in Nicotiana benthamiana (NB) cultivated hydroponically in moving gullies. This type of cultivation system offers great potential for large-scale production of NB plants, a requirement for our partner to the project, Medicago inc., whose objective is to produce each year tens of million doses of quadrivalent seasonal flu vaccines. Medicago has improved the NB expression platform considerably over the last decade. A vaccine antigen, the influenza virus hemagglutinin H1, expressed in leaves of agroinfiltrated NB plants is accumulated in membrane-enveloped virus-like particles, which are later purified by an extraction-clarification process. Our goal will be to establish, for NB plants grown and incubated in a gully system, the best crop management program (i.e. irrigation regime as a function of plant growth, attributes of the nutrient solution, recirculation and disinfection) for maximal production of recombinant proteins. The project will contribute to basic knowledge on this model plant in a context of increased manufacturing capacity, and provide Medicago and other organizations with useful data and guidelines on cultural practices to achieve high recombinant protein yield and quality in NB plants grown hydroponically in gullies. It will also contribute to the training of highly qualified graduate students and personnel in molecular biotechnology, a R&D sector of economic importance in Canada.