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. 2023 Feb 28;10(3):304. doi: 10.3390/bioengineering10030304

Figure 4.

Figure 4

(a) CFPS expression of four IgGs in a 1-mL flowerplate reaction format with either HC/LC co-expression (green bars) or the expression of HC alone with PFLC reagent addition at 0.5 mg/L (blue bars). These reactions were performed in a plate-based format with temperature control but without pH or DO control. (b) CFPS expression of four IgGs in a 250-mL stirred tank reaction format with either HC/LC co-expression (green bars) or the expression of HC alone with PFLC reagent addition at 0.5 mg/L (blue bars). These reactions had temperature, agitation, DO, and two-sided pH control. Data represent mean ± SD for n = 2 titer assays from a single reaction. (c) Comparison of the relative improvement of cell-free expression by flower plate (FP) or Dasbox (DB) observed with the PFLC reagent compared to HC/LC co-expression for the four IgGs tested. The gray bars represent the theoretical titer improvement expected given a constant level of cell-free protein synthesis, folding, and assembly between the two reaction formats. Orange bars represent the additional titer increase observed, which is partially attributed to the LC acting as a co-chaperone to improve HC solubility and final IgG folding and assembly.