Journal: Frontiers in Bioengineering and Biotechnology
Article Title: High-throughput optimization of antibody production in CHO cells by tuning heavy- and light-chain promoter strength
doi: 10.3389/fbioe.2025.1747473
Figure Lengend Snippet: mAb B: effects of LC/HC promoter strength on titer, cell density, and viability post-transfection (72 hpt), and model profiling. (A) Viability (%), (B) viable cell density (VCD; cells/mL), and (C) titer (mg/L) for all LC/HC combinations. Promoter levels: 5, 40, and 100 RPU for both LC and HC. Bars show mean ± SD (n = 2). Statistical analysis was based on one-way ANOVA with Tukey's HSD; different letters indicate p < 0.05. (D) Transfection efficiency (% GFP+) of a control plasmid (pMAX-GFP) quantified by flow cytometry. (E) JMP profiler for least-squares models of titer, VCD, and viability versus LC/HC promoter strength. Left: predicted means with 95% Cls. Right: composite desirability (0-1) balancing high titer with acceptable VCD and viability. Red dashed lines mark targets/constraints.
Article Snippet: CHO-S cells (R80007, Life Technologies, Waltham, MA, United States) CD CHO medium (10743029, Gibco, Thermo Fisher Scientific, Waltham, MA, United States) L-Glutamine (25030081, Thermo Fisher Scientific, Waltham, MA, United States) Freestyle MAX Transfection Reagent (16447100, Thermo Fisher Scientific, Waltham, MA, United States) OptiPRO serum free medium (12309019, Thermo Fisher Scientific, Waltham, MA, United States) Transfection positive control expressing GFP (pMax-E2F1, 16007, Addgene, Watertown, MA, United States) Solution 18 (910-3018, ChemoMetec A/S, Allerød, Denmark) NC-Slide A8 (941-0002, ChemoMetec A/S, Allerød, Denmark)
Techniques: Transfection, Control, Plasmid Preparation, Flow Cytometry