Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment
<i>Centruroides huichol</i> scorpion venom is lethal to mammals. Analysis of the venom allowed the characterization of four lethal toxins named Chui2, Chui3, Chui4, and Chui5. scFv 10FG2 recognized well all toxins except Chui5 toxin, therefore a partial neutralization of the venom was ob...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/6/369 |
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| author | Hugo Valencia-Martínez Timoteo Olamendi-Portugal Rita Restano-Cassulini Hugo Serrano-Posada Fernando Zamudio Lourival D. Possani Lidia Riaño-Umbarila Baltazar Becerril |
| author_facet | Hugo Valencia-Martínez Timoteo Olamendi-Portugal Rita Restano-Cassulini Hugo Serrano-Posada Fernando Zamudio Lourival D. Possani Lidia Riaño-Umbarila Baltazar Becerril |
| author_sort | Hugo Valencia-Martínez |
| collection | DOAJ |
| description | <i>Centruroides huichol</i> scorpion venom is lethal to mammals. Analysis of the venom allowed the characterization of four lethal toxins named Chui2, Chui3, Chui4, and Chui5. scFv 10FG2 recognized well all toxins except Chui5 toxin, therefore a partial neutralization of the venom was observed. Thus, scFv 10FG2 was subjected to three processes of directed evolution and phage display against Chui5 toxin until obtaining scFv HV. Interaction kinetic constants of these scFvs with the toxins were determined by surface plasmon resonance (SPR) as well as thermodynamic parameters of scFv variants bound to Chui5. In silico models allowed to analyze the molecular interactions that favor the increase in affinity. In a rescue trial, scFv HV protected 100% of the mice injected with three lethal doses 50 (LD<sub>50</sub>) of venom. Moreover, in mix-type neutralization assays, a combination of scFvs HV and 10FG2 protected 100% of mice injected with 5 LD<sub>50</sub> of venom with moderate signs of intoxication. The ability of scFv HV to neutralize different toxins is a significant achievement, considering the diversity of the species of Mexican venomous scorpions, so this scFv is a candidate to be part of a recombinant anti-venom against scorpion stings in Mexico. |
| first_indexed | 2024-03-09T22:19:21Z |
| format | Article |
| id | doaj.art-889f7e682bb34a75b4d29e9e68442aad |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T22:19:21Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-889f7e682bb34a75b4d29e9e68442aad2023-11-23T19:16:46ZengMDPI AGToxins2072-66512022-05-0114636910.3390/toxins14060369Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody FragmentHugo Valencia-Martínez0Timoteo Olamendi-Portugal1Rita Restano-Cassulini2Hugo Serrano-Posada3Fernando Zamudio4Lourival D. Possani5Lidia Riaño-Umbarila6Baltazar Becerril7Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoInvestigador por México, CONACyT-Laboratorio de Biología Sintética, Estructural y Molecular, Laboratorio de Agrobiotecnología-Tecnoparque CLQ, Universidad de Colima, Carretera Los Limones-Loma de Juárez, Colima 28627, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoInvestigadora por México, CONACyT-Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca 62250, Mexico<i>Centruroides huichol</i> scorpion venom is lethal to mammals. Analysis of the venom allowed the characterization of four lethal toxins named Chui2, Chui3, Chui4, and Chui5. scFv 10FG2 recognized well all toxins except Chui5 toxin, therefore a partial neutralization of the venom was observed. Thus, scFv 10FG2 was subjected to three processes of directed evolution and phage display against Chui5 toxin until obtaining scFv HV. Interaction kinetic constants of these scFvs with the toxins were determined by surface plasmon resonance (SPR) as well as thermodynamic parameters of scFv variants bound to Chui5. In silico models allowed to analyze the molecular interactions that favor the increase in affinity. In a rescue trial, scFv HV protected 100% of the mice injected with three lethal doses 50 (LD<sub>50</sub>) of venom. Moreover, in mix-type neutralization assays, a combination of scFvs HV and 10FG2 protected 100% of mice injected with 5 LD<sub>50</sub> of venom with moderate signs of intoxication. The ability of scFv HV to neutralize different toxins is a significant achievement, considering the diversity of the species of Mexican venomous scorpions, so this scFv is a candidate to be part of a recombinant anti-venom against scorpion stings in Mexico.https://www.mdpi.com/2072-6651/14/6/369affinity maturationkinetic and thermodynamic parametersphage displayscFvscorpion toxins |
| spellingShingle | Hugo Valencia-Martínez Timoteo Olamendi-Portugal Rita Restano-Cassulini Hugo Serrano-Posada Fernando Zamudio Lourival D. Possani Lidia Riaño-Umbarila Baltazar Becerril Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment Toxins affinity maturation kinetic and thermodynamic parameters phage display scFv scorpion toxins |
| title | Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment |
| title_full | Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment |
| title_fullStr | Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment |
| title_full_unstemmed | Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment |
| title_short | Characterization of Four Medically Important Toxins from <i>Centruroides huichol</i> Scorpion Venom and Its Neutralization by a Single Recombinant Antibody Fragment |
| title_sort | characterization of four medically important toxins from i centruroides huichol i scorpion venom and its neutralization by a single recombinant antibody fragment |
| topic | affinity maturation kinetic and thermodynamic parameters phage display scFv scorpion toxins |
| url | https://www.mdpi.com/2072-6651/14/6/369 |
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