Publikationsdatum:
2024-05-16
Beschreibung:
Snakebite envenoming is a priority Neglected Tropical Disease that causes an estimated 81,000–135,000 fatalities each year. The development of a new generation of safer, affordable, and accessible antivenom therapies is
urgently needed. With this goal in mind, rigorous characterisation of the specific toxins in snake venom is key to
generating novel therapies for snakebite. Monoclonal antibodies directed against venom toxins are emerging as
potentially strong candidates in the development of new snakebite diagnostics and treatment. Venoms comprise
many different toxins of which several are responsible for their pathological effects. Due to the large variability of
venoms within and between species, formulations of combinations of human antibodies are proposed as the next
generation antivenoms. Here a high-throughput screening method employing antibody-based ligand fishing of
venom toxins in 384 filter-well plate format has been developed to determine the antibody target/s The approach
uses Protein G beads for antibody capture followed by exposure to a full venom or purified toxins to bind their
respective ligand toxin(s). This is followed by a washing/centrifugation step to remove non-binding toxins and an
in-well tryptic digest. Finally, peptides from each well are analysed by nanoLC-MS/MS and subsequent Mascot
database searching to identify the bound toxin/s for each antibody under investigation. The approach was
successfully validated to rapidly screen antibodies sourced from hybridomas, derived from venom-immunised
mice expressing either regular human antibodies or heavy-chain-only human antibodies (HCAbs).
Repository-Name:
National Museum of Natural History, Netherlands
Materialart:
info:eu-repo/semantics/article
Format:
application/pdf