Basic cultural determinants of recombinant protein yield in Nicotiana benthamiana used as a transient expression host for the flu vaccine antigen hemagglutinin H1

Authors: Shang, Lingling
Advisor: Michaud, DominiquePépin, Steeve
Abstract: Plants are promising hosts for the production of medically-useful recombinant protein sand numerous studies have been done over the years to optimize transgene expression rates and protein maturation processes in plant systems. By comparison, little is still known about the influence of basic environmental factors and cultural practices on the expression and yield of heterologous proteins in plants. Current cultural practices in greenhouse settings, that generally allow for an increased biomass or food/flower product yield, are not necessarily well suited to recombinant protein production in a molecular farming context. In this study, we investigated the effects of CO2 enrichment, supplemental lighting, ammonium fertigation and plant culture density on growth, development and recombinant protein yield of the protein expression host Nicotiana benthamiana used to express the flu vaccine antigen influenza virus hemagglutinin H1. In brief, our data showed (1) atmospheric CO2 enrichment, high-light irradiance, supplemental LED inter-lighting in the plant canopy and high-ammonium fertigation to enhanced leaf biomass production and endogenous protein content on a plant basis, and (2) LED inter-lighting or elevated plant density to increase recombinant protein yield on a whole-crop area basis. On the other hand, H1 content was not influenced or negatively affected by CO2 enrichment, high-light irradiance or high-ammonium supply on a leaf fresh weight basis. Overall, our findings indicate that the optimal cultural practices for the production of horticultural food products or ornementals in controlled environment settings may not be optimal in molecular farming settings, where the ultimate goal is recombinant protein yield and quality, not leaf biomass, nutrient content, fruit yield or flower quality.
Document Type: Thèse de doctorat
Issue Date: 2019
Open Access Date: 14 November 2019
Grantor: Université Laval
Collection:Thèses et mémoires

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