Zika virus-like particle vaccine fusion loop mutation increases production yield but fails to protect AG129 mice against Zika virus challenge
Fig 2
Comparison of the maturation state of lead and F108A particles by prM/M protein detection and E epitope mapping.
(A) SDS-PAGE followed by Coomassie protein staining of 500ng and 1μg of purified lead and F108A ZIKV VLPs. Molecular weight ladders on this gel are the Novex Sharp (NS; Invitrogen, cat#57318, molecular weights for this ladder shown in kDa) and the Super Signal (SS; Thermo, cat#847868) (B) SDS-PAGE followed by immunoblotting analysis of 500ng and 1μg of purified lead and F108A ZIKV VLPs. An anti-prM polyclonal Ab against ZIKV (Genetex, cat#gtx133305) was used to detect VLP proteins. (C) Binding of each mAb to F108A VLP relative to lead VLP measured by Biolayer Interferometry. Binding was measured at 100 seconds (ZIKV-116, ZV67, IM-79), 250 seconds (ZIKV-117, ZV48, LM-081) or both 100 and 250 seconds (ZIKV-117). For ZIKV-117, only the highest F108A concentration tested fell on the standard curves for the 100 and 250-second measurements, so both are shown. For each Ab, 2 or 4 F108A VLP dilutions fell on the lead VLP standard curve. For those dilutions, the F108A VLP concentration was calculated by least squares fit from the 4PL curve, and the ratios of bound F108A VLP to bound lead VLP were calculated. Refer to S1 Fig for standard curves. Symbols represent the ratios from individual measurements, bars and numbers at the top of each group represent geometric mean ratios, and an equivalence line representing equal mAb binding to F108A and lead is shown at 1.