Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity
Antivenom immunotherapy is the mainstay of treatment for snakebite envenoming. Most parts of the world affected by snakebite envenoming depend on broad-spectrum polyspecific antivenoms that are known to contain a low content of case-specific efficacious immunoglobulins. Thus, advances in toxin-speci...
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MDPI AG
2022-04-01
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| Series: | Toxins |
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| author | Ernest Z. Manson Mutinda C. Kyama Josephine Kimani Aleksandra Bocian Konrad K. Hus Vladimír Petrilla Jaroslav Legáth James H. Kimotho |
| author_facet | Ernest Z. Manson Mutinda C. Kyama Josephine Kimani Aleksandra Bocian Konrad K. Hus Vladimír Petrilla Jaroslav Legáth James H. Kimotho |
| author_sort | Ernest Z. Manson |
| collection | DOAJ |
| description | Antivenom immunotherapy is the mainstay of treatment for snakebite envenoming. Most parts of the world affected by snakebite envenoming depend on broad-spectrum polyspecific antivenoms that are known to contain a low content of case-specific efficacious immunoglobulins. Thus, advances in toxin-specific antibodies production hold much promise in future therapeutic strategies of snakebite envenoming. We report anti-3FTxs monoclonal antibodies developed against <i>N. ashei</i> venom in mice. All the three test mAbs (P4G6a, P6D9a, and P6D9b) were found to be IgG antibodies, isotyped as IgG1. SDS-PAGE analysis of the test mAbs showed two major bands at approximately 55 and 29 kDa, suggestive of immunoglobulin heavy and light chain composition, respectively. The immunoaffinity-purified test mAbs demonstrated higher binding efficacy to the target antigen compared to negative control. Similarly, a cocktail of the test mAbs was found to induce a significantly higher inhibition (<i>p</i>-value < 0.0001) compared to two leading commercial brands of antivenoms on the Kenyan market, implying a higher specificity for the target antigen. Both the test mAbs and 3FTxs polyclonal antibodies induced comparable inhibition (<i>p</i>-value = 0.9029). The inhibition induced by the 3FTxs polyclonal antibodies was significantly different from the two antivenoms (<i>p</i>-value < 0.0001). Our results demonstrate the prospects of developing toxin-specific monoclonal-based antivenoms for snakebite immunotherapy. |
| first_indexed | 2024-03-09T12:54:58Z |
| format | Article |
| id | doaj.art-8352d56a604a4d14aa0332fae89d5f98 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T12:54:58Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-8352d56a604a4d14aa0332fae89d5f982023-11-30T22:01:17ZengMDPI AGToxins2072-66512022-04-0114428510.3390/toxins14040285Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition ActivityErnest Z. Manson0Mutinda C. Kyama1Josephine Kimani2Aleksandra Bocian3Konrad K. Hus4Vladimír Petrilla5Jaroslav Legáth6James H. Kimotho7Institute for Basic Sciences, Technology & Innovation, Pan African University, Nairobi 00100, KenyaDepartment of Medical Laboratory Science, College of Health Sciences, Jomo Kenyatta University of Agriculture & Technology, Nairobi 00100, KenyaDepartment of Biochemistry, College of Health Sciences, Jomo Kenyatta University of Agriculture & Technology, Nairobi 00100, KenyaDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandDepartment of Biology and Physiology, University of Veterinary Medicine and Pharmacy, 041-81 Košice, SlovakiaDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandKenya Medical Research Institute, Nairobi 00100, KenyaAntivenom immunotherapy is the mainstay of treatment for snakebite envenoming. Most parts of the world affected by snakebite envenoming depend on broad-spectrum polyspecific antivenoms that are known to contain a low content of case-specific efficacious immunoglobulins. Thus, advances in toxin-specific antibodies production hold much promise in future therapeutic strategies of snakebite envenoming. We report anti-3FTxs monoclonal antibodies developed against <i>N. ashei</i> venom in mice. All the three test mAbs (P4G6a, P6D9a, and P6D9b) were found to be IgG antibodies, isotyped as IgG1. SDS-PAGE analysis of the test mAbs showed two major bands at approximately 55 and 29 kDa, suggestive of immunoglobulin heavy and light chain composition, respectively. The immunoaffinity-purified test mAbs demonstrated higher binding efficacy to the target antigen compared to negative control. Similarly, a cocktail of the test mAbs was found to induce a significantly higher inhibition (<i>p</i>-value < 0.0001) compared to two leading commercial brands of antivenoms on the Kenyan market, implying a higher specificity for the target antigen. Both the test mAbs and 3FTxs polyclonal antibodies induced comparable inhibition (<i>p</i>-value = 0.9029). The inhibition induced by the 3FTxs polyclonal antibodies was significantly different from the two antivenoms (<i>p</i>-value < 0.0001). Our results demonstrate the prospects of developing toxin-specific monoclonal-based antivenoms for snakebite immunotherapy.https://www.mdpi.com/2072-6651/14/4/285<i>Naja ashei</i>three-finger toxinsmonoclonal antibodiessnakebite envenominginhibition ELISA |
| spellingShingle | Ernest Z. Manson Mutinda C. Kyama Josephine Kimani Aleksandra Bocian Konrad K. Hus Vladimír Petrilla Jaroslav Legáth James H. Kimotho Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity Toxins <i>Naja ashei</i> three-finger toxins monoclonal antibodies snakebite envenoming inhibition ELISA |
| title | Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity |
| title_full | Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity |
| title_fullStr | Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity |
| title_full_unstemmed | Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity |
| title_short | Development and Characterization of Anti-<i>Naja ashei</i> Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity |
| title_sort | development and characterization of anti i naja ashei i three finger toxins 3ftxs specific monoclonal antibodies and evaluation of their in vitro inhibition activity |
| topic | <i>Naja ashei</i> three-finger toxins monoclonal antibodies snakebite envenoming inhibition ELISA |
| url | https://www.mdpi.com/2072-6651/14/4/285 |
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