摘要
目的建立从新鲜猪血中分离纯化血红蛋白的方法,为血液代用品的研究和开发提供原料保障。方法通过离心、微滤和弱阴离子交换层析法,制备高纯度高活性的猪血红蛋白,并建立合理的检验方法(紫外可见光扫描、电泳、高效液相色谱、血凝试验抑制试验和血气分析等),以确定纯化产品的含量、纯度和活性。结果应用以上方法制备的猪血红蛋白的含量为100g/dL;SEHPLC纯度为995%;高铁血红蛋白的含量为08%;残余磷脂的含量为0;血凝抑制试验呈阴性;半饱和氧分压P50为166mmHg;Hill系数为267。结论应用该法制备的猪血红蛋白纯度高活性好,适合作为血液代用品研究和开发用血红蛋白原料。
AIM:To establish a method of hemoglobin isolation and purification from fresh porcine blood. To supply ample and eligible material for the research and development of human blood substitutes. METHOD:Porcine hemoglobin with high purity and activity was prepared by centrifuging,micro filtration and weak anion exchange chromatography. Its properties were analyzed by ultra-visual spectrophotometer scan,electrophoresis,high performance liquid chromatography,hemagglutination inhibition test and blood gas analysis. RESULT: The content of hemoglobin was 10.0 g/dL. The purity by SE-HPLC was 99.5%. The content of methemoglobin was 0.8%. The remains of phosphalipids were 0 ug/mL. Hemagglutination inhibition test was negative. The value of P_~50 was 16.6 mmHg and Hill coefficient was 2.67. CONCLUSION: Porcine hemoglobin prepared through this method had high purity and good allosteric effect and Bohr effect. The method is fit for preparing hemoglobin for the research and development of human blood substitutes.
出处
《中国天然药物》
SCIE
CAS
CSCD
2005年第1期60-64,共5页
