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A Single Tri-Epitopic Antibody Virtually Recapitulates the Potency of a Combination of Three Monoclonal Antibodies in Neutralization of Botulinum Neurotoxin Serotype A
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Toxins 2018, 10(3), 105; https://doi.org/10.3390/toxins10030105

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

1
Zuckerberg San Francisco General Hospital and Trauma Center, Room 3C-38, Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Avenue, San Francisco, CA 94110, USA
2
Knobbe Martin, 2040 Main Street, 14th Floor, Irvine, CA 92614, USA
3
Department of Molecular Biology and Genetics, Faculty of Science, Istanbul Medeniyet University, Unalan Mahallesi, Unalan Sokak, D100 Karayolu, Uskudar-Istanbul 34700, Turkey
4
Molecular and Translational Sciences Division, United States Army Medical Institute of Infectious Diseases (USAMRIID), Fort Detrick, MD 21702, USA
5
Ke’aki Technologies LLC, United States Army Medical Institute of Infectious Diseases, (USAMRIID) Fort Detrick, MD 21702, USA
6
Medical Countermeasures Technology, USAMRIID, Fort Detrick, MD 21702-5011, USA
*
Author to whom correspondence should be addressed.
Received: 12 December 2017 / Revised: 5 February 2018 / Accepted: 13 February 2018 / Published: 1 March 2018
(This article belongs to the Special Issue Botulinum Neurotoxins Antibody and Vaccine)
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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) is in pre-clinical development with an investigational new drug (IND) application filing expected in 2018. View Full-Text
Keywords: botulinum neurotoxin; oligoclonal antibodies; serotype E botulism; recombinant antibodies; antibody engineering; mouse neutralization assay; botulinum antitoxin botulinum neurotoxin; oligoclonal antibodies; serotype E botulism; recombinant antibodies; antibody engineering; mouse neutralization assay; botulinum antitoxin
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Garcia-Rodriguez, C.; Razai, A.; Geren, I.N.; Lou, J.; Conrad, F.; Wen, W.-H.; Farr-Jones, S.; Smith, T.J.; Brown, J.L.; Skerry, J.C.; Smith, L.A.; Marks, J.D. A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes. Toxins 2018, 10, 105.

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