[Preparation and functional identification of human single chain recombinant antibody targeting α-hemolysin of Staphylococcus aureus]

Abstract

Objective To prepare a new fully human antibody against α-hemolysin of Staphylococcus aureus (S. aureus) and to investiagete its neutralizing effect. Methods The IgG-like scFv-Fc inserted into the pcDNA3.1 vector by homologous recombination was expressed in HEK293F suspension cells and purified. ELISA was used to detect the purified scFv538-Fc's binding activity and specificity to S. aureus. The cell proliferation & toxicity assay and rabbit erythrocyte hemolysis assay were used to identify the scFv538-Fc against α-hemolysin of S. aureus. Results A new fully human recombinant antibody scFv-Fc against S. aureus. α-hemolysin was successfully prepared. The mass of the purified scFv-Fc was about 55 kDa. The purified antibody had binding activity to scFv538-Fc, and the antibody bound to Staphylococcus aureus specifically. The results of A549 cytotoxicity assays showed that scFv538-Fc had protective effects on A549 cells. The result of anti-rabbit erythrocyte hemolysis assay confirmed that scFv538-Fc had a significant neutralizing effect on toxins. Conclusion A novel fully human scFv-Fc antibody neutralizing the α-hemolysin toxin of S. aureus is successfully prepared.