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Home / Equine Research Bank / Toxins (Basel) / A Three-Monoclonal Antibody Combination Potently Neutralizes...
Toxins2023; 15(5); 316; doi: 10.3390/toxins15050316

A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice.

Abstract: Equine-derived antitoxin (BAT) is the only treatment for botulism from botulinum neurotoxin serotype G (BoNT/G). BAT is a foreign protein with potentially severe adverse effects and is not renewable. To develop a safe, more potent, and renewable antitoxin, humanized monoclonal antibodies (mAbs) were generated. Yeast displayed single chain Fv (scFv) libraries were prepared from mice immunized with BoNT/G and BoNT/G domains and screened with BoNT/G using fluorescence-activated cell sorting (FACS). Fourteen scFv-binding BoNT/G were isolated with K values ranging from 3.86 nM to 103 nM (median K 20.9 nM). Five mAb-binding non-overlapping epitopes were humanized and affinity matured to create antibodies hu6G6.2, hu6G7.2, hu6G9.1, hu6G10, and hu6G11.2, with IgG K values ranging from 51 pM to 8 pM. Three IgG combinations completely protected mice challenged with 10,000 LDs of BoNT/G at a total mAb dose of 6.25 μg per mouse. The mAb combinations have the potential for use in the diagnosis and treatment of botulism due to serotype G and, along with antibody combinations to BoNT/A, B, C, D, E, and F, provide the basis for a fully recombinant heptavalent botulinum antitoxin to replace the legacy equine product.
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Publication Date: 2023-04-30 PubMed ID: 37235351PubMed Central: PMC10222606DOI: 10.3390/toxins15050316Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper discusses the development of a potent, safe, and renewable antitoxin for botulism using humanized monoclonal antibodies in place of the traditional equine-derived antitoxin.

Introduction

  • The paper presents research aiming to develop a more potent, safer, and renewable antitoxin to treat botulism caused by botulinum neurotoxin serotype G (BoNT/G).
  • The traditional treatment for BoNT/G botulism is the equine-derived antitoxin (BAT), but since it is a foreign protein, it can carry possible severe side effects, and is not renewable.

Methodology

  • The researchers used yeast to display single chain Fv (scFv) libraries made from mice immunized with BoNT/G and BoNT/G domains.
  • The libraries were then screened using BoNT/G via fluorescence-activated cell sorting (FACS).
  • Fourteen scFv with binding properties to BoNT/G were isolated. Their binding strength ranged from 3.86 nM to 103 nM, with a median strength of 20.9 nM.

Antibody Development and Testing

  • Five monoclonal antibodies (mAb) that bind non-overlapping epitopes were developed, then humanized and affinity matured to create the antibodies hu6G6.2, hu6G7.2, hu6G9.1, hu6G10, and hu6G11.2.
  • The strength of these mAbs’ binding was measured, with IgG K values ranging from 51 pM to 8 pM.
  • Three combinations of these mAbs were found to completely protect mice from a challenge with 10,000 lethal doses of BoNT/G. The total mAb dose administered was 6.25 μg per mouse.

Antitoxin Potentials

  • These mAbs combinations provide the potential for diagnosing and treating botulism due to serotype G.
  • Alongside mAbs combinations for BoNT/A, B, C, D, E, and F, this research can form the basis of developing a fully recombinant heptavalent botulinum antitoxin.
  • This new development could be a replacement for the legacy equine product, providing a more potent, safer, and renewable treatment option.

Cite This Article

APA
Fan Y, Lou J, Tam CC, Wen W, Conrad F, Leal da Silva Alves P, Cheng LW, Garcia-Rodriguez C, Farr-Jones S, Marks JD. (2023). A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice. Toxins (Basel), 15(5), 316. https://doi.org/10.3390/toxins15050316

Publication

Toxins
ISSN: 2072-6651
NlmUniqueID: 101530765
Country: Switzerland
Language: English
Volume: 15
Issue: 5
PII: 316

Researcher Affiliations

Fan, Yongfeng
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Lou, Jianlong
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Tam, Christina C
  • Western Regional Research Center, Agricultural Research Station, United States Department of Agriculture, Albany, CA 94710, USA.
Wen, Weihua
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Conrad, Fraser
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Leal da Silva Alves, Priscila
  • Western Regional Research Center, Agricultural Research Station, United States Department of Agriculture, Albany, CA 94710, USA.
Cheng, Luisa W
  • Western Regional Research Center, Agricultural Research Station, United States Department of Agriculture, Albany, CA 94710, USA.
Garcia-Rodriguez, Consuelo
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Farr-Jones, Shauna
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.
Marks, James D
  • Department of Anesthesia and Perioperative Care, University of California, 1001 Potrero Ave., San Francisco, CA 94110, USA.

MeSH Terms

  • Mice
  • Animals
  • Horses
  • Antibodies, Monoclonal
  • Botulism / prevention & control
  • Botulinum Toxins, Type A
  • Single-Chain Antibodies
  • Antitoxins
  • Saccharomyces cerevisiae / metabolism
  • Immunoglobulin G

Conflict of Interest Statement

Authors Marks, Fan, Lou, are named inventors on the patent covering the antibodies described here. The other authors declare no competing interests. The funders had no role in the study design, data collection, analysis, decision to publish, or preparation of the manuscript. Opinions, interpretations, conclusions, and recommendations are those of the authors and not necessarily endorsed by the National Institute of Allergy and Infectious Diseases, or the National Institutes of Health.

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