Non-Destructive Measurement of Target Protein Content in Leaves for Plant-Made Pharmaceutical Protein Production

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In this study, we aimed at developing a non-destructive measurement technique using chlorophyll (Chl) fluorescence to quantify pharmaceutical protein content in leaves in a viral vector-based transient gene expression system using whole plants. A plasmid vector containing a cDNA of hemagglutinin (HA), an influenza vaccine antigen, as well as a tobamoviral replicon, was introduced into Nicotiana benthamiana plants via vacuum-infiltration with the transformed agrobacteria. The HA protein content in leaves increased from 3 d to 6 d after the gene introduction event. Simultaneously, the photosynthetic capacity of leaves markedly declined. It was demonstrated that this decrease in photosynthetic capacity can be detected by measuring Chl fluorescence parameters including the quantum efficiency and thermal dissipation activity of photosystem II, suggesting that the Chl fluorescence measurement can potentially be a useful technique for monitoring the dynamics of HA content in leaves.