Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV
Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that infect pigs’ intestinal epithelial cells, causing high morbidity and mortality. Due to the rapid mutation of PEDV, vaccine efficacy is uncertain, prompting exploration of alternative treatments. Nanobodies, also known as...
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MDPI AG
2024-11-01
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| Series: | Veterinary Sciences |
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| author | Li Zhang Wei Miao Mo Zhou Miao Lin Changyao Fu Zhi Wu Xinnuo Lei Jialong Xu Shinuo Cao Shanyuan Zhu |
| author_facet | Li Zhang Wei Miao Mo Zhou Miao Lin Changyao Fu Zhi Wu Xinnuo Lei Jialong Xu Shinuo Cao Shanyuan Zhu |
| author_sort | Li Zhang |
| collection | DOAJ |
| description | Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that infect pigs’ intestinal epithelial cells, causing high morbidity and mortality. Due to the rapid mutation of PEDV, vaccine efficacy is uncertain, prompting exploration of alternative treatments. Nanobodies, also known as variable heavy chain domains of heavy chain-only antibodies (VHHs), offer significant potential in biomedical applications due to their small size and high specificity. In this study, yeast two-hybrid technology was employed to screen for eight specific VHH sequences targeting the PEDV S protein from a synthetically constructed nanobody yeast library. The VHH genes were then cloned into expression plasmids for recombinant protein production, and the resulting VHHs (termed PEDV S-VHHs) were purified. Indirect immunofluorescence assay (IFA) and Western blotting analysis confirmed that these VHHs specifically bind to both PEDV and its S protein. Neutralization assays demonstrated that seven PEDV S-VHHs exhibited potent neutralizing activity against PEDV. Additionally, a combination of these seven antibodies showed enhanced antiviral effects. Preliminary predictions were also made regarding the binding sites between these VHHs and PEDV. The PEDV S-VHHs described in this study hold potential as candidates for the prevention and treatment of PEDV infection. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-ffeddc6e372e4fd3a6967e39d63c0f092024-11-26T18:24:50ZengMDPI AGVeterinary Sciences2306-73812024-11-01111153310.3390/vetsci11110533Neutralizing VHH Antibodies Targeting the Spike Protein of PEDVLi Zhang0Wei Miao1Mo Zhou2Miao Lin3Changyao Fu4Zhi Wu5Xinnuo Lei6Jialong Xu7Shinuo Cao8Shanyuan Zhu9Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaCollege of Veterinary Pharmacy, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaCollege of Veterinary Pharmacy, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaCollege of Veterinary Pharmacy, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaMedical School, Nanjing University, Nanjing 210093, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaJiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, ChinaPorcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that infect pigs’ intestinal epithelial cells, causing high morbidity and mortality. Due to the rapid mutation of PEDV, vaccine efficacy is uncertain, prompting exploration of alternative treatments. Nanobodies, also known as variable heavy chain domains of heavy chain-only antibodies (VHHs), offer significant potential in biomedical applications due to their small size and high specificity. In this study, yeast two-hybrid technology was employed to screen for eight specific VHH sequences targeting the PEDV S protein from a synthetically constructed nanobody yeast library. The VHH genes were then cloned into expression plasmids for recombinant protein production, and the resulting VHHs (termed PEDV S-VHHs) were purified. Indirect immunofluorescence assay (IFA) and Western blotting analysis confirmed that these VHHs specifically bind to both PEDV and its S protein. Neutralization assays demonstrated that seven PEDV S-VHHs exhibited potent neutralizing activity against PEDV. Additionally, a combination of these seven antibodies showed enhanced antiviral effects. Preliminary predictions were also made regarding the binding sites between these VHHs and PEDV. The PEDV S-VHHs described in this study hold potential as candidates for the prevention and treatment of PEDV infection.https://www.mdpi.com/2306-7381/11/11/533porcine epidemic diarrhea virusspike (S) proteinneutralizing antibodynanobodyVHH |
| spellingShingle | Li Zhang Wei Miao Mo Zhou Miao Lin Changyao Fu Zhi Wu Xinnuo Lei Jialong Xu Shinuo Cao Shanyuan Zhu Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV Veterinary Sciences porcine epidemic diarrhea virus spike (S) protein neutralizing antibody nanobody VHH |
| title | Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV |
| title_full | Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV |
| title_fullStr | Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV |
| title_full_unstemmed | Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV |
| title_short | Neutralizing VHH Antibodies Targeting the Spike Protein of PEDV |
| title_sort | neutralizing vhh antibodies targeting the spike protein of pedv |
| topic | porcine epidemic diarrhea virus spike (S) protein neutralizing antibody nanobody VHH |
| url | https://www.mdpi.com/2306-7381/11/11/533 |
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