Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization
Recombinant monoclonal antibody (mAb) botulinum neurotoxin (BoNT) antitoxins, consisting of three mAbs that bind non-overlapping epitopes, are highly potent. However, the three-mAb mixtures pose unique development and manufacturing challenges. Combining even more mAbs to create multivalent antitoxin...
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MDPI AG
2025-06-01
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| author | Jianlong Lou Wei Hua Wen Fraser Conrad Christina C. Tam Consuelo Garcia-Rodriguez Shauna Farr-Jones James D. Marks |
| author_facet | Jianlong Lou Wei Hua Wen Fraser Conrad Christina C. Tam Consuelo Garcia-Rodriguez Shauna Farr-Jones James D. Marks |
| author_sort | Jianlong Lou |
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| description | Recombinant monoclonal antibody (mAb) botulinum neurotoxin (BoNT) antitoxins, consisting of three mAbs that bind non-overlapping epitopes, are highly potent. However, the three-mAb mixtures pose unique development and manufacturing challenges. Combining even more mAbs to create multivalent antitoxin drugs multiplies those challenges. We previously reported that a single tri-epitopic IgG1-based mAb (TeAb) containing the variable domains of the three parental BoNT/A mAbs and an Fc was as potent as the combination of three IgGs in the mouse neutralization assay (MNA). Here, we extended the tri-epitopic strategy to three other BoNT serotypes. Each TeAb (TeAb-B for BoNT/B, TeAb-E for BoNT/E, and TeAb-F for BoNT/F) binding was measured using fluorescence-activated cell sorting and flow fluorimetry, and the potency was tested in the MNA. The three TeAbs displayed binding affinities that were the same within error of the parental IgGs for each epitope, and all had higher avidity to each serotype of BoNT than that of the parental mAbs. The potency of the BoNT/B, BoNT/E, and BoNT/F TeAbs was similar to the combinations of the three parental IgGs binding BoNT/B, BoNT/E, and BoNT/F in the MNA. We now have four examples of a single TeAb recapitulating the affinity and in vivo potency of a three-mAb antitoxin. The tri-epitopic strategy could be applied to streamline the production and bioanalytics of antibody drugs where three-mAb binding is required for activity. |
| format | Article |
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| issn | 2072-6651 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
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| series | Toxins |
| spelling | doaj-art-909fd4cbe9fb4a1284a7ca8dc9b3657a2025-06-25T14:29:15ZengMDPI AGToxins2072-66512025-06-0117628110.3390/toxins17060281Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin NeutralizationJianlong Lou0Wei Hua Wen1Fraser Conrad2Christina C. Tam3Consuelo Garcia-Rodriguez4Shauna Farr-Jones5James D. Marks6Department of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USADepartment of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USADepartment of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USAFoodborne Toxin Detection and Prevention Research Unit, Western Regional Research Center, Agricultural Research Services, United States Department of Agriculture, 800 Buchanan Street, Albany, CA 94710, USADepartment of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USADepartment of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USADepartment of Anesthesia and Perioperative Care, University of California, Zuckerberg San Francisco General Hospital, Box 1327, 2540 23rd St, San Francisco, CA 94143, USARecombinant monoclonal antibody (mAb) botulinum neurotoxin (BoNT) antitoxins, consisting of three mAbs that bind non-overlapping epitopes, are highly potent. However, the three-mAb mixtures pose unique development and manufacturing challenges. Combining even more mAbs to create multivalent antitoxin drugs multiplies those challenges. We previously reported that a single tri-epitopic IgG1-based mAb (TeAb) containing the variable domains of the three parental BoNT/A mAbs and an Fc was as potent as the combination of three IgGs in the mouse neutralization assay (MNA). Here, we extended the tri-epitopic strategy to three other BoNT serotypes. Each TeAb (TeAb-B for BoNT/B, TeAb-E for BoNT/E, and TeAb-F for BoNT/F) binding was measured using fluorescence-activated cell sorting and flow fluorimetry, and the potency was tested in the MNA. The three TeAbs displayed binding affinities that were the same within error of the parental IgGs for each epitope, and all had higher avidity to each serotype of BoNT than that of the parental mAbs. The potency of the BoNT/B, BoNT/E, and BoNT/F TeAbs was similar to the combinations of the three parental IgGs binding BoNT/B, BoNT/E, and BoNT/F in the MNA. We now have four examples of a single TeAb recapitulating the affinity and in vivo potency of a three-mAb antitoxin. The tri-epitopic strategy could be applied to streamline the production and bioanalytics of antibody drugs where three-mAb binding is required for activity.https://www.mdpi.com/2072-6651/17/6/281tri-epitopic antibodyrecombinant monoclonal antibodybotulinum neurotoxinantitoxinbotulismserotypes B, E, F |
| spellingShingle | Jianlong Lou Wei Hua Wen Fraser Conrad Christina C. Tam Consuelo Garcia-Rodriguez Shauna Farr-Jones James D. Marks Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization Toxins tri-epitopic antibody recombinant monoclonal antibody botulinum neurotoxin antitoxin botulism serotypes B, E, F |
| title | Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization |
| title_full | Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization |
| title_fullStr | Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization |
| title_full_unstemmed | Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization |
| title_short | Single Tri-Epitopic Antibodies (TeAbs) to Botulinum Neurotoxin Serotypes B, E, and F Recapitulate the Full Potency of a Combination of Three Monoclonal Antibodies in Toxin Neutralization |
| title_sort | single tri epitopic antibodies teabs to botulinum neurotoxin serotypes b e and f recapitulate the full potency of a combination of three monoclonal antibodies in toxin neutralization |
| topic | tri-epitopic antibody recombinant monoclonal antibody botulinum neurotoxin antitoxin botulism serotypes B, E, F |
| url | https://www.mdpi.com/2072-6651/17/6/281 |
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