Figure 1. Characterization of anti-SNAP25₁₉₇ monoclonal antibody 2E2A6.

Specificity demonstrated by lack of cross-reactivity towards SNAP25₂₀₆. A. Neuro-2a cells were treated with BoNT/A (150 kDa) from 0.01 to 10 nM (duplicate wells) for 24 h. Western blot was performed with 2E2A6 antibody (1 µg/mL). No cross-reactivity with SNAP25₂₀₆ (no bands observed in the 0 nM BoNT/A lanes) and no other non-specific bands were detected in the whole blot. B & C. Surface plasmon resonance (SPR) was used to characterize the 2E2A6 antibody and to compare the binding affinity and binding kinetics of 2E2A6 and MC-6053. 2E2A6 bound SNAP25₁₉₇ with high affinity and did not bind SNAP25₂₀₆ at any of the concentrations tested up to 10 µM. MC-6053 was able to bind SNAP25₂₀₆ with a K_D of 240 nM. D. The K_d for 2E2A6 is tenfold lower than the K_d for MC-6053 as shown in the normalized graph resulting in better affinity.