Abstract

Background: Treatment of diseases related to enterotoxin B of S. aureus (SEB) is challenging due to the emergence of antibiotic resistance in Staphylococcus aureus (S. aureus). Antibodies specific to SEB can potentially play a role in the treatment of SEB-related illnesses of humans. The aim of the present study was to produce human monoclonal antibodies against recombinant SEB (rSEB) by in vitro immunization of peripheral blood lymphocytes (PBLs). rSEB which was previously purified was used to produce monoclonal antibodies by in vitro immunization of PBLs in the presence of PWM.
Methods: We established a sensitive and reproducible cell culture system where 0.3 µg/µL of rSEB was used to trigger rSEB specific antibodies by PBLs. Furthermore, cloning of cells for monoclonal antibodies was done through limiting dilution.
Results: Supernatants were harvested and assayed for both human polyclonal and monoclonal anti-SEB IgGs respectively by ELISA. All the supernatants from cell culture showed presence of human polyclonal and monoclonal antibodies, whereas specifically 3 wells reported to have higher concentration of monoclonal antibodies.
Conclusions: This is the first testimony on the production of human monoclonal antibodies against rSEB in Pakistan. In future, these antibodies can be used for immunodiagnostic assays as well as immunotherapy of S. aureus.

DOI: 10.7754/Clin.Lab.2025.250122