Fig. 3
Covalent coupling of L1-VLPs. DBCO pre-activated L1-VLPs captured azide-labeled unrelated peptides. Coupling efficiency was analyzed via gel electrophoresis followed by Coomassie staining (A). (B) The antigenicity of L1-DBCO (+) compared to L1 alone (-) was assessed by western blotting (left panel) using an HPV-16 L1 antibody (CAMVIR-1, Santa Cruz Biotechnology, USA). Likewise, L1 was pre-activated with DBCO groups and conjugated to a fluorescent azide-linker to prove the conjugation efficiency. Fluorescently conjugated L1 (+) and a control without DBCO pre-activation (-) were analyzed after SDS-PAGE by fluorescence imaging (right panel). Complete gel images are shown in Supplementary Figure S2. (C) After N3-modified Env trimers were covalently linked to the surface of the L1-VLPs, unbound Env was efficiently separated by SEC.
