Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin

Tetanus toxin (TeNT) is one of the most toxic proteins. Neutralizing antibodies against TeNT are effective in prevention and treatment. In this study, 14 anti-tetanus nanobodies were obtained from a phage display nanobody library by immunizing a camel with the C-terminal receptor-binding domain of T...

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Main Authors: Kexuan Cheng, Jiansheng Lu, Jiazheng Guo, Rong Wang, Lei Chen, Xi Wang, Yujia Jiang, Yating Li, Changyan Xu, Qinglin Kang, Gulisaina Qiaerxie, Peng Du, Chen Gao, Yunzhou Yu, Zhixin Yang, Wei Wang
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Human Vaccines & Immunotherapeutics
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Online Access:https://www.tandfonline.com/doi/10.1080/21645515.2024.2366641
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author Kexuan Cheng
Jiansheng Lu
Jiazheng Guo
Rong Wang
Lei Chen
Xi Wang
Yujia Jiang
Yating Li
Changyan Xu
Qinglin Kang
Gulisaina Qiaerxie
Peng Du
Chen Gao
Yunzhou Yu
Zhixin Yang
Wei Wang
author_facet Kexuan Cheng
Jiansheng Lu
Jiazheng Guo
Rong Wang
Lei Chen
Xi Wang
Yujia Jiang
Yating Li
Changyan Xu
Qinglin Kang
Gulisaina Qiaerxie
Peng Du
Chen Gao
Yunzhou Yu
Zhixin Yang
Wei Wang
author_sort Kexuan Cheng
collection DOAJ
description Tetanus toxin (TeNT) is one of the most toxic proteins. Neutralizing antibodies against TeNT are effective in prevention and treatment. In this study, 14 anti-tetanus nanobodies were obtained from a phage display nanobody library by immunizing a camel with the C-terminal receptor-binding domain of TeNT (TeNT-Hc) as the antigen. After fusion with the human Fc fragment, 11 chimeric heavy-chain antibodies demonstrated nanomolar binding toward TeNT-Hc. The results of toxin neutralization experiments showed that T83–7, T83–8, and T83–13 completely protected mice against 20 × the median lethal dose (LD50) at a low concentration. The neutralizing potency of T83–7, T83–8, and T83–13 against TeNT is 0.4 IU/mg, 0.4 IU/mg and 0.2 IU/mg, respectively. In the prophylactic setting, we found that 5 mg/kg of T83–13 provided the mice with full protection from tetanus, even when they were injected 14 days before exposure to 20 × LD50 TeNT. T83–7 and T83–8 were less effective, being fully protective only when challenged 7 or 10 days before exposure, respectively. In the therapeutic setting, 12 h after exposure to TeNT, 1 ~ 5 mg/kg of T83–7, and T83–8 could provide complete protection for mice against 5 × LD50 TeNT, while 1 mg/kg T83–13 could provide complete protection 24 h after exposure to 5 × LD50 TeNT. Our results suggested that these antibodies represent prophylactic and therapeutic activities against TeNT in a mouse model. The T83–7, T83–8, and T83–13 could form the basis for the subsequent development of drugs to treat TeNT toxicity.
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spelling doaj-art-13b0ffc440814458b9e5816a3a74d3a72025-08-20T02:16:40ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2024-12-0120110.1080/21645515.2024.2366641Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxinKexuan Cheng0Jiansheng Lu1Jiazheng Guo2Rong Wang3Lei Chen4Xi Wang5Yujia Jiang6Yating Li7Changyan Xu8Qinglin Kang9Gulisaina Qiaerxie10Peng Du11Chen Gao12Yunzhou Yu13Zhixin Yang14Wei Wang15Department of Occupational Health and Occupational Diseases, College of Public Health, Zhengzhou University, Zhengzhou, Henan, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaDepartment of Occupational Health and Occupational Diseases, College of Public Health, Zhengzhou University, Zhengzhou, Henan, ChinaDepartment of Occupational Health and Occupational Diseases, College of Public Health, Zhengzhou University, Zhengzhou, Henan, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaLaboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, ChinaDepartment of Occupational Health and Occupational Diseases, College of Public Health, Zhengzhou University, Zhengzhou, Henan, ChinaTetanus toxin (TeNT) is one of the most toxic proteins. Neutralizing antibodies against TeNT are effective in prevention and treatment. In this study, 14 anti-tetanus nanobodies were obtained from a phage display nanobody library by immunizing a camel with the C-terminal receptor-binding domain of TeNT (TeNT-Hc) as the antigen. After fusion with the human Fc fragment, 11 chimeric heavy-chain antibodies demonstrated nanomolar binding toward TeNT-Hc. The results of toxin neutralization experiments showed that T83–7, T83–8, and T83–13 completely protected mice against 20 × the median lethal dose (LD50) at a low concentration. The neutralizing potency of T83–7, T83–8, and T83–13 against TeNT is 0.4 IU/mg, 0.4 IU/mg and 0.2 IU/mg, respectively. In the prophylactic setting, we found that 5 mg/kg of T83–13 provided the mice with full protection from tetanus, even when they were injected 14 days before exposure to 20 × LD50 TeNT. T83–7 and T83–8 were less effective, being fully protective only when challenged 7 or 10 days before exposure, respectively. In the therapeutic setting, 12 h after exposure to TeNT, 1 ~ 5 mg/kg of T83–7, and T83–8 could provide complete protection for mice against 5 × LD50 TeNT, while 1 mg/kg T83–13 could provide complete protection 24 h after exposure to 5 × LD50 TeNT. Our results suggested that these antibodies represent prophylactic and therapeutic activities against TeNT in a mouse model. The T83–7, T83–8, and T83–13 could form the basis for the subsequent development of drugs to treat TeNT toxicity.https://www.tandfonline.com/doi/10.1080/21645515.2024.2366641Tetanus toxinnanobodyheavy chain antibodyneutralizing antibodyphage display library
spellingShingle Kexuan Cheng
Jiansheng Lu
Jiazheng Guo
Rong Wang
Lei Chen
Xi Wang
Yujia Jiang
Yating Li
Changyan Xu
Qinglin Kang
Gulisaina Qiaerxie
Peng Du
Chen Gao
Yunzhou Yu
Zhixin Yang
Wei Wang
Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
Human Vaccines & Immunotherapeutics
Tetanus toxin
nanobody
heavy chain antibody
neutralizing antibody
phage display library
title Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
title_full Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
title_fullStr Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
title_full_unstemmed Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
title_short Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin
title_sort characterization of neutralizing chimeric heavy chain antibodies against tetanus toxin
topic Tetanus toxin
nanobody
heavy chain antibody
neutralizing antibody
phage display library
url https://www.tandfonline.com/doi/10.1080/21645515.2024.2366641
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