Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells
The Taiwanese cobra, <i>Naja atra</i>, is a clinically significant species of snake observed in the wild in Taiwan. Victims bitten by <i>N. atra</i> usually experience severe pain and local tissue necrosis. Although antivenom is available for treatment of cobra envenomation,...
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MDPI AG
2022-07-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/7/459 |
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| author | Chien-Chun Liu Cho-Ju Wu Tsai-Ying Chou Geng-Wang Liaw Yung-Chin Hsiao Lichieh-Julie Chu Chi-Hsin Lee Po-Jung Wang Cheng-Hsien Hsieh Chun-Kuei Chen Jau-Song Yu |
| author_facet | Chien-Chun Liu Cho-Ju Wu Tsai-Ying Chou Geng-Wang Liaw Yung-Chin Hsiao Lichieh-Julie Chu Chi-Hsin Lee Po-Jung Wang Cheng-Hsien Hsieh Chun-Kuei Chen Jau-Song Yu |
| author_sort | Chien-Chun Liu |
| collection | DOAJ |
| description | The Taiwanese cobra, <i>Naja atra</i>, is a clinically significant species of snake observed in the wild in Taiwan. Victims bitten by <i>N. atra</i> usually experience severe pain and local tissue necrosis. Although antivenom is available for treatment of cobra envenomation, its neutralization potency against cobra-induced necrosis is weak, with more than 60% of cobra envenoming patients developing tissue necrosis after antivenom administration. The present study found that cytotoxin (CTX) is a key component of <i>N. atra</i> venom responsible for cytotoxicity against myoblast cells. Anti-CTX IgY was generated in hens, and the spleens of these hens were used to construct libraries for the development of single chain variable fragments (scFv). Two anti-CTX scFv, S1 and 2S7, were selected using phage display technology and biopanning. Both polyclonal IgY and monoclonal scFv S1 reacted specifically with CTX in cobra venom. In a cell model assay, the CTX-induced cytolytic effect was inhibited only by monoclonal scFv S1, not by polyclonal IgY. Moreover, the neutralization potency of scFv S1 was about 3.8 mg/mg, approximately three times higher than that of conventional freeze-dried neurotoxic antivenom (FNAV). Collectively, these results suggest that scFv S1 can effectively neutralize CTX-induced cytotoxicity and, when combined with currently available antivenom, can improve the potency of the latter, thereby preventing tissue damage induced by cobra envenoming. |
| first_indexed | 2024-03-09T05:46:02Z |
| format | Article |
| id | doaj.art-5576f4ec29dd43459f02be8072a9b982 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T05:46:02Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-5576f4ec29dd43459f02be8072a9b9822023-12-03T12:21:05ZengMDPI AGToxins2072-66512022-07-0114745910.3390/toxins14070459Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 CellsChien-Chun Liu0Cho-Ju Wu1Tsai-Ying Chou2Geng-Wang Liaw3Yung-Chin Hsiao4Lichieh-Julie Chu5Chi-Hsin Lee6Po-Jung Wang7Cheng-Hsien Hsieh8Chun-Kuei Chen9Jau-Song Yu10Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, TaiwanDepartment of Emergency Medicine, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Taoyuan 33305, TaiwanGraduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, TaiwanDepartment of Emergency Medicine, Yeezen General Hospital, Taoyuan 32645, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 33302, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 33302, TaiwanSchool of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 11042, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 33302, TaiwanDepartment of Emergency Medicine, En Chu Kong Hospital, New Taipei City 23741, TaiwanDepartment of Emergency Medicine, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Taoyuan 33305, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 33302, TaiwanThe Taiwanese cobra, <i>Naja atra</i>, is a clinically significant species of snake observed in the wild in Taiwan. Victims bitten by <i>N. atra</i> usually experience severe pain and local tissue necrosis. Although antivenom is available for treatment of cobra envenomation, its neutralization potency against cobra-induced necrosis is weak, with more than 60% of cobra envenoming patients developing tissue necrosis after antivenom administration. The present study found that cytotoxin (CTX) is a key component of <i>N. atra</i> venom responsible for cytotoxicity against myoblast cells. Anti-CTX IgY was generated in hens, and the spleens of these hens were used to construct libraries for the development of single chain variable fragments (scFv). Two anti-CTX scFv, S1 and 2S7, were selected using phage display technology and biopanning. Both polyclonal IgY and monoclonal scFv S1 reacted specifically with CTX in cobra venom. In a cell model assay, the CTX-induced cytolytic effect was inhibited only by monoclonal scFv S1, not by polyclonal IgY. Moreover, the neutralization potency of scFv S1 was about 3.8 mg/mg, approximately three times higher than that of conventional freeze-dried neurotoxic antivenom (FNAV). Collectively, these results suggest that scFv S1 can effectively neutralize CTX-induced cytotoxicity and, when combined with currently available antivenom, can improve the potency of the latter, thereby preventing tissue damage induced by cobra envenoming.https://www.mdpi.com/2072-6651/14/7/459cobra venom<i>Naja atra</i>cytotoxin (CTX)cytotoxicitynecrosissingle-chain variable fragment (scFv) |
| spellingShingle | Chien-Chun Liu Cho-Ju Wu Tsai-Ying Chou Geng-Wang Liaw Yung-Chin Hsiao Lichieh-Julie Chu Chi-Hsin Lee Po-Jung Wang Cheng-Hsien Hsieh Chun-Kuei Chen Jau-Song Yu Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells Toxins cobra venom <i>Naja atra</i> cytotoxin (CTX) cytotoxicity necrosis single-chain variable fragment (scFv) |
| title | Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells |
| title_full | Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells |
| title_fullStr | Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells |
| title_full_unstemmed | Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells |
| title_short | Development of a Monoclonal scFv against Cytotoxin to Neutralize Cytolytic Activity Induced by <i>Naja atra</i> Venom on Myoblast C2C12 Cells |
| title_sort | development of a monoclonal scfv against cytotoxin to neutralize cytolytic activity induced by i naja atra i venom on myoblast c2c12 cells |
| topic | cobra venom <i>Naja atra</i> cytotoxin (CTX) cytotoxicity necrosis single-chain variable fragment (scFv) |
| url | https://www.mdpi.com/2072-6651/14/7/459 |
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