Development of a monoclonal antibody against a synthetic peptide of Buthotus saulcyi scorpion venom: a novel diagnostic and neutralizing tool.

Background: Hybridoma technology is an essential method used to produce monoclonal antibodies, providing equal specificity and scalability for biomedical applications. This methodology utilizes a hybridoma formation process through fusing short-lived, antibody-producing B lymphocytes with immortalized myeloma cells, thus providing hybridoma clones that produce monoclonal antibodies that are highly specific. Monoclonal antibodies produced through hybridoma technology have a consistent and reproducibility benefits over polyclonal forms of antibodies, making monoclonal antibodies an essential product in diagnostics and therapeutics. In this study, we attempted to produce monoclonal antibodies in order to target a synthetic peptide from venom of Buthotus Saulcyi, a medically important scorpion in the family Buthidae, native to Iran, known for having potent toxicity that is most dangerous in children and the elderly.

Methods: Balb/c mice were immunized with the synthetic peptide P4 before fusion to Sp2/0-Ag14 myeloma cells using polyethylene glycol at a 5:1 ratio. Hybridoma cells were cultured in HAT selective media with a single clone isolated using limiting dilution.

Result: Cell production was confirmed with an enzyme-linked immunosorbent assay (ELISA) and determined specificity to recognize B. Saulcyi venom. Neutralization was determined using MTT and SRB cell lines HepG2 and determined the monoclonal antibody treatment for B. Saulcyi venom had efficacy.

Conclusion: These findings highlight the potential of this mAb as a diagnostic tool for rapid detection of B. Saulcyi venom in clinical settings, paving the way for improved management of scorpion envenomation.