Fig. 3 | Nature Communications

Fig. 3

From: Structure and immunogenicity of a stabilized HIV-1 envelope trimer based on a group-M consensus sequence

Fig. 3

Design and characterization of ConM SOSIP-ferritin nanoparticles. a Left: linear representation of ConM SOSIP.v7-ferritin. The N-terminus from H. pylori ferritin (purple) is linked to the SOSIP.v7 C-terminus by a Gly-Ser-Gly (GSG) linker. Right: model of eight ConM SOSIP.v7 trimers with gp120 subunits (red) and gp41 subunits (pink), displayed on the H. pylori ferritin nanoparticle (purple, PDB: 3BVE) viewed down the two-fold, three-fold, and four-fold axis of the ferritin particle, respectively. The figure was drawn using PyMol. b The two-dimensional class averages of the negative-stain electron microscopic analyses of ConM SOSIP.v7-ferritin nanoparticle. Example images of the micrographs are shown in Supplementary Fig. 5c. c Representative dynamic light scattering graphs showing the size distribution of ConM SOSIP.v7 trimer and SOSIP.v7-ferritin nanoparticle. d Glycan profile of the ConM SOSIP.v7-ferritin nanoparticle as determined by hydrophilic interaction liquid chromatography-ultra performance liquid chromatography. The circle diagram indicates the relative amounts of Man5–9GlcNac2 (M5–M9) and complex glycans. e Thermostability (measured by differential scanning calorimetry) of PGT145-purified ConM SOSIP.v7-ferritin nanoparticle expressed in 293F cells. The ConM SOSIP.v7 trimer melting curve (Fig. 1e) is shown in red for comparison. f Binding of broadly neutralizing antibodies (bNAbs) 2G12 and PGT145 and non-neutralizing antibodies (non-NAbs) F105 and 19b to ConM SOSIP.v7 trimer and SOSIP.v7-ferritin (100 nM SOSIP.v7 equivalent each) measured by biolayer interferometry after incubation at different temperatures. Shown are the mean values of the area under the curve (AUC) derived from the binding curves in Supplementary Fig. 6. g Calcium flux (relative fluorescence) in B cells expressing either VRC01, PGT145, PGT121, or PG16 as the B cell receptor, stimulated with ConM SOSIP.v7 trimers or ConM SOSIP.v7-ferritin nanoparticles

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