Figure 1.

Generation of the Ab-VAMP₇₇ antibody. (A) Scheme showing the three SNARE proteins involved in neuroexocytosis that form the SNARE complex by coil-coiling their SNARE motifs, one from the vesicular VAMP-1/2 (blue), two from the membrane-anchored SNAP-25 (pink), and one from the integral membrane protein syntaxin-1A/1B (red). (B) Scheme showing the peptide bonds cleaved by the VAMP-specific CNT used in this study, which generate specific new N-termini to VAMP (shown is human VAMP-2). (C) Alignment of VAMP-1/2 showing that the FETSAAKLKRKYW peptide (pink) exposed by BoNT/B and TeNT is in all the main animal species used in research. The green residues indicate cleavage sites of TeNT and BoNT/B in VAMP-1 and VAMP-2. Red residues indicate the mutation responsible for VAMP-1 resistance to TeNT and BoNT/B in rat and chicken. (D) Scheme showing the generation of immunogenic carrier by chemical conjugation of the C-terminal Cysteine to the Keyhole limpet Hemocyanin. (E) the peptide-KLH complex was injected into a rabbit, and at the scheduled time point the blood was collected for the ensuing purification of peptide-specific IgGs.