Isolation and fractionation of ferritin from human term placenta--a source for human isoferritins

A method for isolating ferritin from human term placenta was described. The placenta was homogenized in water containing protease inhibitor and heated at 70 degrees C. The ferritin was precipitated with ammonium sulfate at pH 5.2 and purified by repeated cycles of ultracentrifugation and molecular sieve chromatography through Sepharose 4B columns. Isoelectric focusing revealed a broad spectrum of isoferritins. These isoferritins were separated by ion-exchange chromatography on Sephadex A-25 at pH 7.5 and stepwise elution with increasing concentrations of NaCl. By this method "basic," "intermediate," and "acid" isoferritins were separated. The most basic placental isoferritin was shown to be identical to splenic ferritin by isoelectric focusing, subunit analysis, and fluorescent ELISA. The acid placental isoferritin had similar characteristics to heart-type ferritin. It was suggested that the easily available placental tissue could serve as a source for human isoferritins in research and in clinical assays.