Bioinformatic analysis and characterization of the anti-CD20 scFv-conjugated Fe3O4 nanoparticles for targeted therapy of CD20+ leukemia cells.

OBJECTIVES: Chronic lymphocytic leukemia (CLL) is one of the common types of leukemia. Various approaches for CLL therapy have been advanced, but they have some adverse effects. Iron oxide (Fe3O4) nanoparticles are promising drug carriers due to their magnetic properties and biocompatibility. MATERIALS AND METHODS: The recombinant anti-CD20 scFv was computationally modeled using the AlphaFold server. The complementarity-determining regions (CDR) of scFv were identified using the IMGT/V-Quest database. Molecular docking was performed separately between the scFv antibody and the CD20 antigen (ClusPro 2.0) and between Fe3O4 and the scFv (PyRx). Then, synthesized Fe₃O₄ nanoparticles were characterized using X-ray diffraction (XRD), field-emission scanning electron microscopy (FE-SEM), dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FTIR). In vitro biological evaluation of the complexes was performed using the MTT assay. RESULTS: The molecular docking studies revealed favorable interactions between the recombinant scFv antibody and the CD20 antigen. Fe3O4 nanoparticles were included as a simplified ligand model to explore possible surface interaction patterns with the scFv molecule, suggesting no predicted interactions with CDR regions in the docking model. Physicochemical characterization confirmed the successful synthesis and desirable structural attributes of the Fe₃O₄ nanoparticles. In vitro biological evaluation of the Fe₃O₄ nanoparticle--anti-CD20 scFv complex revealed that cell death in the Raji cell line was significantly higher than in the control groups, underscoring the targeted cytotoxic efficacy. CONCLUSION: These findings suggest that the Fe3O4 nanoparticle complex conjugated to an anti-CD20 scFv antibody can serve as a novel strategy for CLL treatment, contributing to the development of targeted therapies for cancer.