血红蛋白交联条件优化及其携氧性能

Characterization of cross-linked bovine hemoglobin and its oxygen-carrying properties

纯化牛血获得的高纯度血红蛋白,在不同条件下进行戊二醛交联反应得到聚合牛血红蛋白,应用SDS-PAGE对其分子量分布进行分析比较。利用葡萄糖-葡萄糖氧化酶系统和黄嘌呤-黄嘌呤氧化酶系统,研究氧化应激状态下过氧化氢和超氧化物对血红蛋白携氧性能的影响。结果表明,戊二醛交联血红蛋白反应的最佳条件为:溶液总体系5 m L、血红蛋白质量浓度82 g/L、磷酸钾缓冲液(p H=7.4)的浓度为0.1 mol/L、0.47 mol/L的赖氨酸0.2 m L、0.25 mol/L的冷戊二醛1.2 m L及在4℃下150 r/min摇荡24 h;葡萄糖-葡萄糖氧化酶和黄嘌呤-黄嘌呤氧化酶的存在会使血红蛋白被氧化,体系中存在抗氧化剂超氧化物歧化酶(SOD)和过氧化氢酶(CAT),会有效减弱血红蛋白氧化程度,提高其携氧性能。

Taking the high-purity bovine hemoglobin from purified bovine blood as raw materials,we obtained the polymerized bovine hemoglobin polymer through glutaraldehyde cross-linking reaction under different conditions,and analyzed its molecular weight distribution using SDS-PAGE. By use of glucose-glucose oxidase system and xanthine-xanthine oxidase system,the effects of hydrogen peroxide and superoxide on the oxygen-carrying property of bovine hemoglobin were studied in the state of oxidative stress. It is shown that the optimum reaction conditions are： 82 g / L hemoglobin,0. 2 m L of 0. 47 mol / L lysine,1. 2 m L of 0. 25 mol / L glutaraldehyde,0. 1 mol / L KPB at p H 7. 4 in a total volume of 5 m L,at 4 ℃,rocking at 150 r / min for 24 h. Hemoglobin is oxidized under oxidation stress when hydrogen peroxide,glucose and glucose oxidase,xanthine and xanthine oxidase are present. However,the degree of oxidation of hemoglobin is alleviated effectively when antioxidants SOD and catalase exist in the reaction system.