Figure 3.

Scalability of RIT expression to differentiated plants and preparative PCPs (pPCPs). (a) Comparison of PCPs and differentiated N. benthamiana plants for the expression of RITs with different payloads. (b) Accumulation of ER-targeted N-terminal (red dots) and C-terminal (green dots) VisA-based RITs in differentiated N. benthamiana plants according to the incubation time after infiltration (4–6 days). Dotted lines represent 95% confidence intervals (CI) for the model with a fair fit (Table S2) using a natural log transformation. (c) Accumulation of RITs in N. benthamiana plants (different subcellular compartments) for different payloads conjugated to an anti-CD64 scFv. (d) Correlation of DsRed in differentiated N. benthamiana plants and pPCPs generated from a BY-2 culture volume of 10 mL with a cell density of 200 g L⁻¹ (m v⁻¹) and n ≥ 7 biological replicates (N = 24). Lines represent linear fits (Table S3). Error bars represent standard deviation with n ≥ 9 (a) or n ≥ 3 (c) and N = 554 (a) or N = 77 (c) biological replicates. In a and c, statistical significance was determined by two-way ANOVA with Bonferroni correction. Samples without significant differences are marked with identical letters. Apo, apoplast; Chlo, chloroplast; Cyto, cytosol; ER, endoplasmic reticulum; N.B., N. benthamiana.