title: Production and characterization of a murine monoclonal antibody against human ferritin.
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 (2.34×10(9) 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.
  
  
  CONCLUSIONS
  This mAb can specifically recognize ferritin and may serve as a component of ferritin diagnostic kit if other requirements of the kit are met.
authors.affiliation: Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors.forename: Ali Ahmad
authors.initials: AA
authors.lastname: Bayat
authors.forename: Omid
authors.initials: O
authors.lastname: Yeganeh
authors.forename: Roya
authors.initials: R
authors.lastname: Ghods
authors.forename: Amir Hassan
authors.initials: AH
authors.lastname: Zarnani
authors.forename: Reza Bahjati
authors.initials: RB
authors.lastname: Ardekani
authors.forename: Ahmad Reza
authors.initials: AR
authors.lastname: Mahmoudi
authors.forename: Jafar
authors.initials: J
authors.lastname: Mahmoudian
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Haghighat-Noutash
authors.forename: Mahmood
authors.initials: M
authors.lastname: Jeddi-Tehrani
keywords: ELISA; Ferritin; Flow cytometry; Monoclonal antibody; Western blotting