Study On Purification Of Porcine Hemoglobin By Ultrafiltration Membrane Separation

Because of the worldwide shortage of blood resources, emergency surgery, and earthquakes etc., blood substitutes become more and more important. As the main material of blood substitutes, hemoglobin's purity and activity are closely associated with the security of blood substitutes. Ideal hemoglobin material should have high purity,high concentration and natural activity similar to red blood cells(RBCs). There have been many reports of methods of hemoglobin purification recently, however, the activity of hemoglobin is more or less damaged after purification.Ultrafiltration is a membrane separation technique of little damage.According to this characteristic, ultrafiltration is used in our study to separate hemogloblin with high purity and activity close to natural RBCs.About our study:Objective:Establish and optimize the ultrafiltration condition of hemoglobin purification and obtain porcine hemogloblin solution with high purity, high concentration and oxygen carrying capacity close to natural RBCs,which is demanded by the artificial red blood cell research.Methods:The hemoglobin solution was obtained by hypotonic hemolysis from porcine blood cells,and then purified by ultrafiltration. According to the relationship between the permeation flux and the operating pressure together with the feed liquid temperature,select the best condition of ultrafiltration and then purify the porcine hemoglobin. Analyze the purified hemoglobin using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and ultra-visable spectrophotometer. Besides, mensurate the oxygen carrying capacity and P50 of the purified hemoglobin solution with Hemox-analyzer.Results:During the process, permeation flux increases with the operating pressure linearly, and the optimized conditions are as follows: operating pressure is 0.05Mpa, feed liquid temperature is 12℃. The purity of the porcine hemoglobin separated under this conditon reaches over 99%, and the concentrtion is magnified over 20 times.Determined by the hemox-analyzer, hemoglobin purified by ultrafiltration has a"S"curve close to natural RBCs , and P50 is 20.42.Conclusion:The porcine hemoglobin purified by ultrafiltration has not only high concentrtion and high purity, but also oxygen carrying capacity close to natural RBCs, which proves to have little damage to hemoglobin.Our study supports the further research of blood subsitutes with a good material.Ultrafiltration is a commonly used membrane separation technology, with advantages of no pollution, mild contidion ,little protein denature and high productivity. For above reasons, our method of purfying hemoglobin by ultrafiltration may develop into industrialization.