Production and Characterization of a Murine Monoclonal Antibody Against Human Ferritin

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.