In this study, we investigated the efficiency of LNPs loaded with GFP-encoding pDNA using several previously published ionizable lipids, including DLin-MC3-DMA (MC3), CIN-16645 (LP-01), ALC-0315, 4A3-SC8, and DOTAP (SORT), SM-102, and β-sitosterol as a cholesterol analogue in SM-102. Key formulation characteristics, such as polydispersity index (PDI), Z-average diameter (Z-Avg), encapsulation efficiency (%EE), and freeze-thaw stability, were examined through plate-based methods. Our results show that all LNP formulations achieved ≥85% encapsulation of pDNA-eGFP with PDI values ≤0.10, indicating efficient nanoparticle formation. Next, we evaluated the transfection efficiency and mean fluorescence intensity (MFI) of pDNA-loaded LNPs in Huh-7 hepatocytes and A549 lung epithelial cells. We show that pDNA-eGFP-loaded LNPs exhibited comparable GFP intensities, with slightly lower transfection efficiencies than mRNA-loaded LNPs. Both Huh-7 and A549 cells were successfully transfected with pDNA-encapsulated LNPs

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To cite this poster: Konopka, S., Stewart, S., Gronevelt, J.P., et al. In vivo investigation of pDNA delivery using lipid nanoparticle formulations in A549 and Huh7 cell lines. Poster presented at: Cell Bio 2024; December 14-18, San Diego, CA.