Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice
The bacterium <i>Clostridium botulinum</i> is the causative agent of botulism—a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel <i>C. botulinum</i> immunotherapeutics, camelid...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-08-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/11/8/464 |
| _version_ | 1811304419727245312 |
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| author | Svetlana A. Godakova Anatoly N. Noskov Irina D. Vinogradova Galina A. Ugriumova Andrey I. Solovyev Ilias B. Esmagambetov Amir I. Tukhvatulin Denis Y. Logunov Boris S. Naroditsky Dmitry V. Shcheblyakov Aleksandr L. Gintsburg |
| author_facet | Svetlana A. Godakova Anatoly N. Noskov Irina D. Vinogradova Galina A. Ugriumova Andrey I. Solovyev Ilias B. Esmagambetov Amir I. Tukhvatulin Denis Y. Logunov Boris S. Naroditsky Dmitry V. Shcheblyakov Aleksandr L. Gintsburg |
| author_sort | Svetlana A. Godakova |
| collection | DOAJ |
| description | The bacterium <i>Clostridium botulinum</i> is the causative agent of botulism—a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel <i>C. botulinum</i> immunotherapeutics, camelid single-domain antibodies (sdAbs, VHHs, or nanobodies) could be used due to their unique structure and characteristics. In this study, VHHs were produced using phage display technology. A total of 15 different monoclonal VHHs were selected based on their comlementarity-determining region 3 (CDR3) sequences. Different toxin lethal dose (LD<sub>50</sub>) challenges with each selected phage clone were conducted in vivo to check their neutralizing potency. We demonstrated that modification of neutralizing VHHs with a human immunoglobulin G (IgG)1 Fc (fragment crystallizable) fragment (fusionbody, VHH-Fc) significantly increased the circulation time in the blood (up to 14 days). At the same time, VHH-Fc showed the protective activity 1000 times higher than monomeric form when challenged with 5 LD<sub>50</sub>. Moreover, VHH-Fcs remained protective even 14 days after antibody administration. These results indicate that this VHH-Fc could be used as an effective long term antitoxin protection against botulinum type A. |
| first_indexed | 2024-04-13T08:06:48Z |
| format | Article |
| id | doaj.art-5bc5f87f181d46978be593eb2f389dee |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-13T08:06:48Z |
| publishDate | 2019-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-5bc5f87f181d46978be593eb2f389dee2022-12-22T02:55:08ZengMDPI AGToxins2072-66512019-08-0111846410.3390/toxins11080464toxins11080464Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in MiceSvetlana A. Godakova0Anatoly N. Noskov1Irina D. Vinogradova2Galina A. Ugriumova3Andrey I. Solovyev4Ilias B. Esmagambetov5Amir I. Tukhvatulin6Denis Y. Logunov7Boris S. Naroditsky8Dmitry V. Shcheblyakov9Aleksandr L. Gintsburg10Department of Genetics and Bacteria Molecular Biology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Bacteriology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Bacteriology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Bacteriology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Bacteriology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Genetics and Bacteria Molecular Biology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Medical Microbiology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Medical Microbiology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Genetics and Bacteria Molecular Biology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Genetics and Bacteria Molecular Biology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaDepartment of Genetics and Bacteria Molecular Biology, Gamaleya Research Center of Epidemiology and Microbiology, 18 Gamaleya Street, Moscow 123098, RussiaThe bacterium <i>Clostridium botulinum</i> is the causative agent of botulism—a severe intoxication caused by botulinum neurotoxin (BoNT) and characterized by damage to the nervous system. In an effort to develop novel <i>C. botulinum</i> immunotherapeutics, camelid single-domain antibodies (sdAbs, VHHs, or nanobodies) could be used due to their unique structure and characteristics. In this study, VHHs were produced using phage display technology. A total of 15 different monoclonal VHHs were selected based on their comlementarity-determining region 3 (CDR3) sequences. Different toxin lethal dose (LD<sub>50</sub>) challenges with each selected phage clone were conducted in vivo to check their neutralizing potency. We demonstrated that modification of neutralizing VHHs with a human immunoglobulin G (IgG)1 Fc (fragment crystallizable) fragment (fusionbody, VHH-Fc) significantly increased the circulation time in the blood (up to 14 days). At the same time, VHH-Fc showed the protective activity 1000 times higher than monomeric form when challenged with 5 LD<sub>50</sub>. Moreover, VHH-Fcs remained protective even 14 days after antibody administration. These results indicate that this VHH-Fc could be used as an effective long term antitoxin protection against botulinum type A.https://www.mdpi.com/2072-6651/11/8/464camelid single-domain antibodiesVHH<i>Clostridium botulinum</i>toxin neutralizationphage displaydimersFc fragments |
| spellingShingle | Svetlana A. Godakova Anatoly N. Noskov Irina D. Vinogradova Galina A. Ugriumova Andrey I. Solovyev Ilias B. Esmagambetov Amir I. Tukhvatulin Denis Y. Logunov Boris S. Naroditsky Dmitry V. Shcheblyakov Aleksandr L. Gintsburg Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice Toxins camelid single-domain antibodies VHH <i>Clostridium botulinum</i> toxin neutralization phage display dimers Fc fragments |
| title | Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice |
| title_full | Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice |
| title_fullStr | Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice |
| title_full_unstemmed | Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice |
| title_short | Camelid VHHs Fused to Human Fc Fragments Provide Long Term Protection Against Botulinum Neurotoxin A in Mice |
| title_sort | camelid vhhs fused to human fc fragments provide long term protection against botulinum neurotoxin a in mice |
| topic | camelid single-domain antibodies VHH <i>Clostridium botulinum</i> toxin neutralization phage display dimers Fc fragments |
| url | https://www.mdpi.com/2072-6651/11/8/464 |
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