Production and characterization of monoclonal antibodies to synthetic human ferroportin peptide

Background. Ferroptosis is a type of non-apoptotic iron-dependent form of regulated cell death. Ferroportin is the only exporter of iron from cells, however, the involvement of ferroportin in cell ferroptosisis poorly understood. Activation or blocking of ferroptosis pathways may provide a therapeutic strategy for the treatment of a number of diseases. Control of ferroportin expression will make it possible to select a more effective tumor therapy. Obtaining monoclonal antibodies to human ferroportin will help to further study its role in ferroptosis of tumor cells. Aim. Obtaining and characterization of monoclonal antibodies to human ferroportin. Materials and methods. Mouse monoclonal antibodies against a synthetic peptide corresponding to the ferroportin protein region were obtained using hybridomic technology. The peptide was synthesized by the method of solid-phase automatic synthesis. Results. A 15-membered peptide with the sequence ELKQLNLHKDTEPKP (ELK) was selected and synthesized. Conjugates of the synthetic peptide with carrier proteins were obtained. 4 stable hybridomic clones were obtained, and samples of monoclonal antibodies were obtained. The samples were successfully tested using methods of flow cytometry, enzyme immunoassay, and polyacrylamide gel electrophoresis. Conclusion. Four antibodies against ferroportin binding the ELK peptide were obtained and characterized. The antibodies can be used in further studies of iron transport in tumor cells.