Purification Of Human Umbilical Cord Blood Hemoglobin And Its Self - Oxidation Experiment

Blood transfusion is one of the most common treatments in the clinic. Because of the shortage of blood supply, matching complex and creating cross-infection easily, looking for a safe and effective blood substitutes is a key and hot point in the development of medical research in a century. Blood substitute has not the function of carrying oxygen, but also expanding. So it must has the traits such as abundant source,without toxicity and immunegenicity, good mechanical rheological properties, long shelf life and be easy physiological metabolism. It can be split into two categories:Hemoglobin based oxygen carriers and Perfluorocarbon compounds.TO study with the Hemoglobin based oxygen carriers, the product was purified by Q300 anion-exchange chromatography, then the final products was got by concentrating the purified hemoglobin. The concentration of hemoglobin and the purity was analyzed by SDS-PAGE and HPLC, the oxygen saturation curve was analyzed through the blood analyzer.The main ingredients of HBOCs were over pure hemoglobin. Because of the loss of superoxide dismutase (SOD), peroxidase (CAT) and other active enzymes, the system is easy to produce reactive oxygen species (ROS). This article will be reducing the ferulic acid modification to the hemoglobin to inhibit HBOCs oxidation, determination of ferulic acid modified degrees, through 2-IT increased hemoglobin surface free thiol, and joined the sodium azide, xanthine oxidation-xanthine oxidase medium conditions, to explore the modification of ferulic acid degree and its oxidation resistance.The results show that:(1) The concentration of final products was 11.5g/dl, and the content of methemoglobin is 1% or less; hemoglobin had a higher purity by the analysis of SDS-PAGE and HPLC, excluding mixed protein, the purity was 99.10%±0.70% analyzed by Gel Imaging System; the oxygenation of hemoglobin curve had no significant deviation after purification, the activity of carrying oxygen would not be affected. (2) After ferulic acid modified hemoglobin structure and activity of carrying oxygen is not affected, along with the increase of the degree of ferulic acid modified hemoglobin from oxidation speed slower. Its antioxidant capacity is proportional to the ferulic acid modification degree.