The ovotransferrin was extracted and purified from chicken egg. The iron binding capacity of ovotransferrin was studied with different concentration of Fe3+ or NaHCO3， pH value and temperature. The effects of freeze-drying and spray-drying on the physical and chemical properties of ovotransferrin-Fe（Ⅲ） were investigated by colorimeter， infrared spectrometer， particle size analyzer and scanning electron microscope， etc. The iron binding capacity of ovotransferrin was good （capacity 91.5%， iron content 1.28 μg/g）. In the condition of 0.4 mmol/L FeCl3 pH 8.0； 120 mmol/L NaHCO3 and temperature below 60 ℃. There were significant differences in the chromaticity， particle size distribution or complete dissolution time of freeze-drying ovotransferrin-Fe（Ⅲ） and spray-drying ovotransferrin-Fe（Ⅲ）， but there was no significant difference in water activity or chemical bon. After 30 days of animal experiments， freeze-drying and spray-drying group had significant effects on the increase in body weight and organ index in the anemic rats， as well as the red blood cell count， hemoglobin concentration， hematocrit， erythrocyte （P<0.01）. Both groups can significantly increase serum iron content， serum ferritin content， and decrease total iron binding capacity （P<0.05）. Some indexes of ovotransferrin group were even better than ferrous sulfate control group. Therefore， ovotransferrin can be used as a biological iron supplement， which can significantly improve the condition of rat iron deficiency anemia.