Figure 1 | Scientific Reports

Figure 1

From: Antibodies from Sierra Leonean and Nigerian Lassa fever survivors cross-react with recombinant proteins representing Lassa viruses of divergent lineages

Figure 1

Recombinant LASV proteins representing lineages II–IV and their use in ELISA. Panel (A) Nucleoproteins (NP) from LASV representing lineages II–IV were expressed in E. coli, purified and resolved by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Vertical lines indicate removal of lanes from a single gel. Circles: minor contaminants. Asterisk: monomeric NP. Square: NP breakdown product. Panel (B): LASV Zinc protein was expressed in E. coli, purified and resolved by SDS-PAGE and analyzed by western blotting. Monomeric and multimeric forms of the protein (asterisks) are detected. Panel (C) LASV linked glycoprotein was expressed in Drosophila S2 cells, and purified and resolved by SDS-PAGE in the presence and absence of dithiothreitol (DTT). Because most of the GP1 and 2 subunits are covalently linked, reduction by DTT does not affect the mobility of the GP protomer. Panel (D): LASV prefusion glycoprotein (Pf-GP) was expressed in Drosophila S2 cells, purified and resolved by SDS-PAGE in the presence and absence of dithiothreitol (DTT). Reduction by DTT converts GP protomer to GP1 and 2 subunits that have similar molecular weights. Panels (C,D) were prepared from three separate gels. Uncropped polyacrylamide gels and the western blot used to prepare panels (A–D) are shown in Fig. S1. Panel (E): ReLASV Pan-Lassa NP ELISA configured with a mixture of recombinant NP from LASV representing lineages II–IV (LII–IV) and Pan-Lassa Pf-GP ELISA configured with a mixture of recombinant Pf-GP from LASV representing lineages II–IV were used to compare reactivity of IgG and IgM from control samples from heathy United States blood donors (n = 28) versus and Sierra Leonean Lassa fever survivors (n = 58). Data was analyzed using Prism (version 6.07, GraphPad Software, Inc., San Diego, CA) and the figure was compiled using Adobe Illustrator (version 15.1.0, San Jose, CA). Box and whisker plots representing means and standard deviations of samples are indicated. P values for pairwise comparisons are indicated.

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