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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Main Authors: 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
Format: Article
Language:English
Published: MDPI AG 2022-07-01
Series:Toxins
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Online Access:https://www.mdpi.com/2072-6651/14/7/459
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Summary: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.
ISSN:2072-6651