A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes

Toxins
Consuelo Garcia-RodriguezJames D Marks

Abstract

Human botulism is most commonly caused by botulinum neurotoxin (BoNT) serotypes A, B, and E. For this work, we sought to develop a human monoclonal antibody (mAb)-based antitoxin capable of binding and neutralizing multiple subtypes of BoNT/E. Libraries of yeast-displayed single chain Fv (scFv) antibodies were created from the heavy and light chain variable region genes of humans immunized with pentavalent-toxoid- and BoNT/E-binding scFv isolated by Fluorescence-Activated Cell Sorting (FACS). A total of 10 scFv were isolated that bound one or more BoNT/E subtypes with nanomolar-level equilibrium dissociation constants (KD). By diversifying the V-regions of the lead mAbs and selecting for cross-reactivity, we generated three scFv that bound all four BoNT/E subtypes tested at three non-overlapping epitopes. The scFvs were converted to IgG that had KD values for the different BoNT/E subtypes ranging from 9.7 nM to 2.28 pM. An equimolar combination of the three mAbs was able to potently neutralize BoNT/E1, BoNT/E3, and BoNT/E4 in a mouse neutralization assay. The mAbs have potential utility as therapeutics and as diagnostics capable of recognizing multiple BoNT/E subtypes. A derivative of the three-antibody combination (NTM-1633) i...Continue Reading

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Citations

Apr 6, 2019·Current Opinion in Infectious Diseases·Michael P MotleyBettina C Fries
Sep 25, 2019·Toxins·Christine Rasetti-Escargueil, Michel R Popoff
Feb 9, 2020·International Journal of Molecular Sciences·Chih-Hung WangCheng-Yen Kao
Nov 30, 2018·Toxins·Jianlong Lou, James D Marks
Oct 9, 2019·Antimicrobial Agents and Chemotherapy·Doris M SnowMilan T Tomic
Jul 20, 2019·Toxins·Robert J HobbsChristopher D Gwenin
Apr 6, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Sébastien BrierMichel R Popoff

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Methods Mentioned

BETA
Fluorescence-Activated Cell Sorting
FACS
flow cytometry
X-ray
PCR

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