Purification of iron-rich swine hemoglobin and its characteristics

Swine hemoglobin was extracted from fresh lysed swine red blood cell. The lysed solution was precipitated by 10%,20%,30%,40%,50%,and 60% saturated ammonium sulfate,and precipitaton from each step was collected and the cruel swine hemoglobin was obtained. The results showed the impure protein was mainly distributed in 10%~40% saturated ammonium sulfate pelleted precipitations, while the swine hemoglobin was mainly distributed in the precipitation obtained from 50% and 60% saturated ammonium sulfate precipitated production and was separated by the DEAE-Sepharose FF anion material and SDS-PAGE was used to identify their purification. The results of DEAE-Sepharose FF showed that both of the separated dilution got single protein peak. The SDS-PAGE results showed that the hemoglobin was mainly concentrated by 50% and 60% saturated ammonium sulfate.The molecular weight of the purified hemoglobin was 16.4 ku which was in accordance with with that of the monomer of hemoglobin. The purity of the purified hemoglobin was 95.2% canalyzed by PDQuest,which was higer than that of the purchased porcine hemoglobin whose purity was 85%. The protein concentration from each step was determined by Bradford method. The result showed that obtained rate of hemoglobin was accounted for 49.69% in the total protein fractions. The solubility of purified porcine hemoglobin was 32.96%. The iron content in purifiend hemoglobin was 0.335% whicn was closed to the theory content of natural hemoglobin.