Author/Authors :
Bayat، Ali Ahmad نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Yeganeh، Omid نويسنده Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Ghods، Roya نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Zarnani، Amir Hassan نويسنده Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran , , Bahjati Ardekani، Reza نويسنده Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR , , Mahmoudi، Ahmad Reza نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Mahmoudian، Jafar نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Haghighat-Noutash، Farzaneh نويسنده Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Jeddi Tehrani]، Mahmood نويسنده ,
Abstract :
Background: Ferritin is an iron storage protein, which plays a key role in iron metabolism. Measurement of ferritin level in serum is one of the most useful indicators of iron status and also a sensitive measurement of iron deficiency. Monoclonal antibodies may be useful as a tool in various aspects of ferritin investigations. In this paper, the production of a murine monoclonal antibody (mAb) against human ferritin was reported.
Methods: Balb/c mice were immunized with purified human ferritin and splenocytes of hyper immunized mice were fused with Sp2/0 myeloma cells. After four times of cloning by limiting dilution, a positive hybridoma (clone: 2F9-C9) was selected by ELISA using human ferritin. Anti-ferritin mAb was purified from culture supernatants by affinity chromatography.
Results: Determination of the antibody affinity for ferritin by ELISA revealed a relatively high affinity (2.34×109 M-1) and the isotype was determined to be IgG2a. The anti-ferritin mAb 2F9-C9 reacted with 79.4% of Hela cells in flow cytometry. The antibody detected a band of 20 kDa in K562 cells, murine and human liver lysates, purified ferritin in Western blot and also ferritin in human serum.
Conclusion: This mAb can specifically recognize ferritin and may serve as a component of ferritin diagnostic kit if other requirements of the kit are met.
