Monoclonal Antibodies that Inhibit the Proteolytic Activity of Botulinum Neurotoxin Serotype/B

Toxins
Yongfeng FanJ D Marks

Abstract

Existing antibodies (Abs) used to treat botulism cannot enter the cytosol of neurons and bind to botulinum neurotoxin (BoNT) at its site of action, and thus cannot reverse paralysis. However, Abs targeting the proteolytic domain of the toxin could inhibit the proteolytic activity of the toxin intracellularly and potentially reverse intoxication, if they could be delivered intracellularly. As such, antibodies that neutralize toxin activity could serve as potent inhibitory cargos for therapeutic antitoxins against botulism. BoNT serotype B (BoNT/B) contains a zinc endopeptidase light chain (LC) domain that cleaves synaoptobrevin-2, a SNARE protein responsible for vesicle fusion and acetylcholine vesicle release. To generate monoclonal Abs (mAbs) that could reverse paralysis, we targeted the protease domain for Ab generation. Single-chain variable fragment (scFv) libraries from immunized mice or humans were displayed on yeast, and 19 unique BoNT/B LC-specific mAbs isolated by fluorescence-activated cell sorting (FACS). The equilibrium dissociation constants (KD) of these mAbs for BoNT/B LC ranged from 0.24 nM to 14.3 nM (mean KD 3.27 nM). Eleven mAbs inhibited BoNT/B LC proteolytic activity. The fine epitopes of selected mAbs were...Continue Reading

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Citations

Mar 9, 2017·Expert Opinion on Drug Discovery·Marco Pirazzini, Ornella Rossetto
Sep 25, 2019·Toxins·Christine Rasetti-Escargueil, Michel R Popoff
Oct 9, 2019·Antimicrobial Agents and Chemotherapy·Doris M SnowMilan T Tomic
Jun 8, 2017·Toxicon : Official Journal of the International Society on Toxinology·Lisa C Shriver-LakeGeorge P Anderson

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Methods Mentioned

BETA
flow cytometry
Fluorescence
FRET
PCR
electrophoresis

Software Mentioned

GraphPad Prism
Pymol

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