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. 2022 Jun 9;18(6):e1010573. doi: 10.1371/journal.ppat.1010573

Fig 2. Production and characterization of LS-POWV-EDIII nanoparticle immunogen.

Fig 2

(A) Schematic representation of LS-POWV-EDIII nanoparticle. Structures of LS (PDB: 1HQK; green) and POWV EDIII (blue) are covalently linked via SpyCatcher/SpyTag (cyan). (B) Coomassie stained SDS-PAGE gel of POWV EDIII nanoparticle conjugation using SpyCatcher/SpyTag system. Purified POWV EDIII-SpyT (12.6 kDa), LS-SpyC (32 kDa), and conjugated LS-POWV-EDIII (45 kDa) are shown. (C) Size exclusion chromatogram of LS-POWV-EDIII. Sample was run on Sephacryl S-400 HR AKTA column. A retention time of 45 mL was observed (MW 1x106-1x107 Da). (D) ELISA reactivity of mAbs B2 toward POWV EDIII-SpyT and LS-POWV-EDIII. ELISAs were performed twice independently in triplicate wells (mean ± SD). (E) Negative-stain transmission EM images of LS-POWV-EDIII. Scale bar is 50 nm.