A Highly Specific Monoclonal Antibody for Botulinum Neurotoxin Type A-Cleaved SNAP25

Toxins

Catherine Rhບume²  Brian B. Cai²  Joanne Wang²  Ester Fernández-Salas¹  K. Roger Aoki²  Joseph Francis²  Ron S. Broide² 

[1] Department of Pathology, School of Medicine, University of Michigan, Ann Arbor, MI 48109, USA; E-Mail:;Department of Biological Sciences, Allergan, Irvine, CA 92612, USA; E-Mails:

关键词: botulinum neurotoxin;    SNAP25;    neuromuscular junction;    motor nerve;    autonomic nerve;    recombinant antibody;   

DOI  :  10.3390/toxins7072354

来源: mdpi

【 摘 要 】

Botulinum neurotoxin type-A (BoNT/A), as onabotulinumtoxinA, is approved globally for 11 major therapeutic and cosmetic indications. While the mechanism of action for BoNT/A at the presynaptic nerve terminal has been established, questions remain regarding intracellular trafficking patterns and overall fate of the toxin. Resolving these questions partly depends on the ability to detect BoNT/A’s location, distribution, and movement within a cell. Due to BoNT/A’s high potency and extremely low concentrations within neurons, an alternative approach has been employed. This involves utilizing specific antibodies against the BoNT/A-cleaved SNAP25 substrate (SNAP25₁₉₇) to track the enzymatic activity of toxin within cells. Using our highly specific mouse monoclonal antibody (mAb) against SNAP25₁₉₇, we generated human and murine recombinant versions (rMAb) using specific backbone immunoglobulins. In this study, we validated the specificity of our anti-SNAP25₁₉₇ rMAbs in several different assays and performed side-by-side comparisons to commercially-available and in-house antibodies against SNAP25. Our rMAbs were highly specific for SNAP25₁₉₇ in all assays and on several different BoNT/A-treated tissues, showing no cross-reactivity with full-length SNAP25. This was not the case with other reportedly SNAP25₁₉₇-selective antibodies, which were selective in some, but not all assays. The rMAbs described herein represent effective new tools for detecting BoNT/A activity within cells.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.