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

该项目的总体目标是确定调控重组疫苗抗原和其他生物药物在移动沟渠中水培栽培烟草(Nictiana benthamiana(NB))瞬时表达和积累的主要因素。这种类型的培养系统为大规模生产NB植物提供了巨大的潜力，这是我们该项目的合作伙伴Medicago Inc.的要求，该公司的目标是每年生产数千万剂四价季节性流感疫苗。在过去的十年中，Medicago大大改进了NB表达平台。一种疫苗抗原，流感病毒血凝素H1，在农业渗透的NB植物的叶片中表达，在被膜包裹的病毒样颗粒中积累，后来通过提取-澄清过程进行纯化。我们的目标将是为在沟渠系统中生长和培养的NB植物建立最好的作物管理计划(即根据植物生长、营养液的属性、再循环和消毒的函数的灌溉制度)，以最大限度地生产重组蛋白。该项目将有助于在提高制造能力的背景下了解这一模式植物的基本知识，并为Medicago和其他组织提供有用的数据和栽培实践指南，以实现在沟壑中以水培方式种植的NB植物的高重组蛋白产量和质量。它还将有助于培养高素质的分子生物技术研究生和人员，分子生物技术是加拿大具有重要经济意义的研发部门。

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.