Fig. 1: Design and characterization of NiV G nanoparticle.

a Schematic of NiV G nanoparticle (NiV G-ferritin), soluble NiV G head domain (NiV sG), and ferritin proteins. b SEC of NiV G-ferritin, NiV sG, and ferritin on Superose 6 Increase 10/300GL. c SDS-PAGE analysis of NiV G-ferritin, NiV sG, and ferritin under reduced (left) and non-reduced (right) conditions. The gels were derived from the same experiment and processed in parallel. d Size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) analysis of NiV G-ferritin and NiV sG. The SEC-MALS shows the protein component molecular weight (blue line) of NiV G-ferritin and NiV sG is ~1.5 MDa and 48.8 kDa, respectively. e Negative-stain electron micrographs and 2D classification averages of purified NiV G-ferritin and ferritin nanoparticle immunogens. Scale bar, 100 nm. f Hydrodynamic diameter distribution of NiV G-ferritin, NiV sG, and ferritin measured by dynamic light scattering (DLS). Each protein was performed three times. The data represent the mean intensity from three replications. g Thermal stability assay of NiV G-ferritin and NiV sG by differential scanning fluorimetry (DSF). Protein denaturation was monitored as the sample temperature increased. Each protein was performed four times, and representative data are shown. h Antigenicity detection of NiV G-ferritin nanoparticle using published NiV G mAbs by ELISA. HENV-26, HENV-32, and nAH1.3 are NiV G-specific mAbs targeting three distinct epitopes on NiV G. Rabies virus monoclonal antibody RVC20 was used as a negative control. The ELISA was performed twice independently, and representative binding curves are shown. Each symbol represents mean ± SEM.