Production and characterization of monoclonal antibodies to synthetic human ferroportin peptide

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Abstract

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.

About the authors

Margarita G. Khotuleva

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

Author for correspondence.
Email: khotulevamargarita@gmail.com
ORCID iD: 0009-0008-6104-5233

Margarita Gennadievna Khotuleva

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Yana O. Sadovskaya

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0009-0009-7115-7797

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Anastasia O. Karimova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia; National Research University Higher School of Economics

ORCID iD: 0009-0000-0317-9948

24 Kashirskoe Shosse, Moscow 115522

20 Myasnitskaya St., Moscow 101000

Russian Federation

Nikolay E. Varlamov

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0000-0002-8602-6599

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Dmitriy A. Khochenkov

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0000-0002-5694-3492

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Maria V. Mironova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0000-0002-6598-873X

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Dmitriy V. Gusev

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0000-0003-0218-8265

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

Olga N. Solopova

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia

ORCID iD: 0000-0002-5465-6094

24 Kashirskoe Shosse, Moscow 115522

Russian Federation

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