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. 2021 Jul 6;21(13):5697–5705. doi: 10.1021/acs.nanolett.1c01421

Figure 1.

Figure 1

Size of pDNA/PEI particles dictates their transfection efficiency in LVV production cells. (a) Schematic of preparation of pDNA/PEI particles and transfection process for production of LVVs. Each exclamation mark indicates a critical process control parameter influencing transfection outcomes. (b) Transfection efficiencies, characterized as transgene expression levels of the luciferase reporter, in a monolayer culture of HEK293T cells as a function of pDNA concentration at the mixing step and incubation time (10 s to 60 min) before dosage. For the group of mixing at a DNA concentration of 5, 10, or 20 μg/mL, the particles were diluted 5, 10, or 20 times, respectively, to 1 μg/mL to dose cells. (c) Growth in the average size (z-average diameter given by dynamic light scattering, DLS) of pDNA/PEI particles following mixing of pDNA and PEI solutions in Opti-MEM. The growth kinetics is dependent on the concentration of pDNA. The error bars were derived from three independent experiments, demonstrating reproducibility and predictability under the experimental conditions used. (d) Direct correlation between transfection efficiency and the z-average particle size based on data points from all experiments from (b) and (c) with varying pDNA concentrations and incubation times.