A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice
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 KD values ranging from 3.86 nM to 103 nM (median KD 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 KD values ranging from 51 pM to 8 pM. Three IgG combinations completely protected mice challenged with 10,000 LD50s 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.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:15 |
|---|---|
| Enthalten in: |
Toxins - 15(2023), 5 vom: 30. Apr. |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Fan, Yongfeng [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 29.05.2023 Date Revised 30.05.2023 published: Electronic Citation Status MEDLINE |
|---|
| doi: |
10.3390/toxins15050316 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM357376226 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM357376226 | ||
| 003 | DE-627 | ||
| 005 | 20231226072446.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.3390/toxins15050316 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1191.xml |
| 035 | |a (DE-627)NLM357376226 | ||
| 035 | |a (NLM)37235351 | ||
| 035 | |a (PII)316 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Fan, Yongfeng |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 29.05.2023 | ||
| 500 | |a Date Revised 30.05.2023 | ||
| 500 | |a published: Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 KD values ranging from 3.86 nM to 103 nM (median KD 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 KD values ranging from 51 pM to 8 pM. Three IgG combinations completely protected mice challenged with 10,000 LD50s 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a antibody engineering | |
| 650 | 4 | |a botulinum neurotoxin | |
| 650 | 4 | |a mouse neutralization assay | |
| 650 | 4 | |a oligoclonal antibodies | |
| 650 | 4 | |a recombinant antibodies | |
| 650 | 4 | |a serotype G botulism | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Botulinum Toxins, Type A |2 NLM | |
| 650 | 7 | |a EC 3.4.24.69 |2 NLM | |
| 650 | 7 | |a Single-Chain Antibodies |2 NLM | |
| 650 | 7 | |a Antitoxins |2 NLM | |
| 650 | 7 | |a Immunoglobulin G |2 NLM | |
| 700 | 1 | |a Lou, Jianlong |e verfasserin |4 aut | |
| 700 | 1 | |a Tam, Christina C |e verfasserin |4 aut | |
| 700 | 1 | |a Wen, Weihua |e verfasserin |4 aut | |
| 700 | 1 | |a Conrad, Fraser |e verfasserin |4 aut | |
| 700 | 1 | |a Leal da Silva Alves, Priscila |e verfasserin |4 aut | |
| 700 | 1 | |a Cheng, Luisa W |e verfasserin |4 aut | |
| 700 | 1 | |a Garcia-Rodriguez, Consuelo |e verfasserin |4 aut | |
| 700 | 1 | |a Farr-Jones, Shauna |e verfasserin |4 aut | |
| 700 | 1 | |a Marks, James D |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Toxins |d 2009 |g 15(2023), 5 vom: 30. Apr. |w (DE-627)NLM199735913 |x 2072-6651 |7 nnns |
| 773 | 1 | 8 | |g volume:15 |g year:2023 |g number:5 |g day:30 |g month:04 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.3390/toxins15050316 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 15 |j 2023 |e 5 |b 30 |c 04 | ||