Development of Nanobodies Against Hemorrhagic and Myotoxic Components of Bothrops atrox Snake Venom

• Published in: Frontiers in immunology Vol. 11; p. 655
• Main Authors: Bailon Calderon, Henri, Yaniro Coronel, Verónica Olga, Cáceres Rey, Omar Alberto, Colque Alave, Elizabeth Gaby, Leiva Duran, Walter Jhon, Padilla Rojas, Carlos, Montejo Arevalo, Harrison, García Neyra, David, Galarza Pérez, Marco, Bonilla, César, Tintaya, Benigno, Ricciardi, Giulia, Smiejkowska, Natalia, Romão, Ema, Vincke, Cécile, Lévano, Juan, Celys, Mary, Lomonte, Bruno, Muyldermans, Serge
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 07-05-2020
• Subjects: Animals; Antivenins - therapeutic use; Bothrops - metabolism; Camelids, New World - immunology; Crotalid Venoms - chemistry; Crotalid Venoms - immunology; Hemorrhage - drug therapy; hemorrhagic; Immunization - methods; Immunologic Factors - therapeutic use; Immunology; Male; Mice; myotoxic; Myotoxicity - drug therapy; nanobodies; neutralization; Single-Domain Antibodies - therapeutic use; snake; Snake Bites - drug therapy; Treatment Outcome; venom; nanobodies; venom; myotoxic; neutralization; Bothrops atrox; hemorrhagic; snake; viperidae
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2020.00655

Snake envenoming is a globally neglected public health problem. Antivenoms produced using animal hyperimmune plasma remain the standard therapy for snakebites. Although effective against systemic effects, conventional antivenoms have limited efficacy against local tissue damage. In addition, potential hypersensitivity reactions, high costs for animal maintenance, and difficulties in obtaining batch-to-batch homogeneity are some of the factors that have motivated the search for innovative and improved therapeutic products against such envenoming. In this study, we have developed a set of nanobodies (recombinant single-domain antigen-binding fragments from camelid heavy chain-only antibodies) against
snake venom hemorrhagic and myotoxic components. An immune library was constructed after immunizing a
with whole venom of
, from which nanobodies were selected by phage display using partially purified hemorrhagic and myotoxic proteins. Biopanning selections retrieved 18 and eight different nanobodies against the hemorrhagic and the myotoxic proteins, respectively.
assays in mice showed that five nanobodies inhibited the hemorrhagic activity of the proteins; three neutralized the hemorrhagic activity of whole
venom, while four nanobodies inhibited the myotoxic protein. A mixture of the anti-hemorrhagic and anti-myotoxic nanobodies neutralized the local tissue hemorrhage and myonecrosis induced by the whole venom, although the nanobody mixture failed to prevent the venom lethality. Nevertheless, our results demonstrate the efficacy and usefulness of these nanobodies to neutralize important pathologies of the venom, highlighting their potential as innovative therapeutic agents against envenoming by
, a viperid species causing many casualties in South America.