Fig. 1

Design and generation of receptor-antibody hybrid proteins. A - Schematic representation of the receptor-antibody hybrid proteins featuring the decoyMET moiety at the N-terminus and the single chain MET antibody at the C-terminus (DecAb, left panel), or the single chain antibody moiety at the N-terminus and the decoyMET at the C-terminus (AbDec, right panel). SEMA, SEMA domain of MET (light blue: blades 1–4, corresponding to MET α-chain; magenta: blades 5–7). The black arrow indicates the disulfide bond linking α- and β-chains of MET; PSI, Plexin/Semaphorin/Integrin domain of MET. IPT, Immunoglobulin-like/Plexin/Transcription Factor domain of MET. The red star indicates a single point mutation (K842E) inserted in decoyMET to abrogate MvDN30 binding. VL-CL, Variable Light and Constant Light chains of MvDN30; VH-CH1, Variable Heavy and first Constant Heavy chains of MvDN30. The linker between the two moieties of the fusion molecule is highlighted in yellow. B - Coomassie Blue staining of affinity-purified hybrid proteins resolved by SDS-PAGE under reducing (RED) or non-reducing (NON-RED) conditions. In reducing conditions, DecAb hybrid proteins generate two bands of about 40 kDa (MET α-chain) and 150 kDa (MET β-chain + linker + single chain antibody), while AbDec hybrid proteins generate two bands of about 110 kDa (single chain antibody + linker + MET α-chain) and 80 kDa (MET β-chain)