Identification of a common epitope in knottins and phospholipases D present in Loxosceles sp venom by a monoclonal antibody
2023
Alves-Mondini, Camila | Beltramino, Martina | Jiacomini, Isabella, Gizzi | Karim-Silva, Sabrina | dos Santos Antunes, Nicolle | de Moura, Juliana | Aubrey, Nicolas | Billiald, Philippe | Machado-De-Ávila, Ricardo, Andrez | Alvarenga, Larissa, M | Becker-Finco, Alessandra | Laboratorio de Imunoquimica, Departamento de Patologia Basica ; Universidade Federal do Paraná (UFPR) | Infectiologie et Santé Publique (UMR ISP) ; Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Laboratoire de Recherche Vasculaire Translationnelle (LVTS (UMR_S_1148 / U1148)) ; Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité)-Université Sorbonne Paris Nord | Universidade do Extremo Sul Catarinense (UNESC) | This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq, Brazil (No. 472460/2013-7, No. 308314/2020-4), through the Instituto Nacional de Ciência e Tecnologia em Venenos e Antivenenos (INCT-INOVATOX, grant no. 406816/2022-0); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001 and CAPES-PRINT. | ANR-10-LABX-0053,MAbImprove,Optimization of therapeutic monoclonal antibodies development Better antibodies, better developed AND better used(2010)
International audience
Show more [+] Less [-]English. In the Americas and specially in Brazil, the Loxosceles intermedia, Loxosceles gaucho and Loxosceles laeta are the three most medically relevant brown spider species, and whose bites can lead to the condition known as loxoscelism. Here, we report the development of a tool capable of identifying a common epitope amongst Loxosceles sp. venom's toxins. A murine monoclonal antibody (LmAb12) and its recombinant fragments (scFv12P and diabody12P) have been produced and characterized. This antibody and its recombinant constructs were able to recognize proteins of Loxosceles spider venoms with specificity. The scFv12P variant was also able to detect low concentrations of Loxosceles venom in a competitive ELISA assay, displaying potential as a venom identification tool. The primary antigenic target of LmAb12 is a knottin, a venom neurotoxin, that has a shared identity of 100 % between the L. intermedia and L. gaucho species and high similarity to L. laeta. Furthermore, we observed LmAb12 was able to partially inhibit in vitro hemolysis, a cellular event typically induced by the Loxosceles sp. venoms. Such behavior might be due to LmAb12 cross-reactivity between the antigenic target of LmAb12 and the venom's dermonecrotic toxins, the PLDs, or even the existence of synergism between these two toxins.
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