Fig. 3: Schematic of rational design and characterization of LNP-based DNA vaccine formulation.
From: Nanoparticle-based DNA vaccine protects against SARS-CoV-2 variants in female preclinical models
A Traditional four-component LNPs, which consist of ionizable lipid component combined with a phospholipid to support bilayer structure, cholesterol to enhance bilayer stability, and a lipid-anchored-PEG to increase circulation time prepared via microfluidic mixing method. Representative cryo-TEM microscopy of LNPs encapsulated with pDNA: B LNP-C C LNP-HPS (scale bar, 200 nm). D Average hydrodynamic diameter and polydispersity index (PDI) of LNPs, measured by dynamic light scattering (DLS) (n = 6/group). E Zeta potential showed slightly negatively charged LNPs. pKa values of these LNPs varied from 7 to 7.4 (n = 6/group). F Agarose gels of LNPs after DNase activity assay reveals preserved pDNA when encapsulated in LNPs. G, H Spike protein expression in HEK-293 cells treated with LNP-C and LNP-HPS after 48 h via immunofluorescence (n = 5/group). Representative fluorescence images of HEK-293 cells marked with anti-Spike antibody. Samples were stained with anti-SARS-CoV-2 Spike protein (RBD) antibody (magenta), Phalloidin (green), and DAPI (blue), acquired using a ×10 objective. Data are presented as mean ± SEM; ****p < 0.0001. G One-way ANOVA followed by Tukey’s multiple comparison test. B–H Each experiment was repeated at least three times independently with similar results and the representative dataset is presented. Source data are provided as a Source Data file.
