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Mérieux公司自1950年开始用胎盘提取血浆蛋白。每个胎盘含有大约200ml血液,这就意味着每年节省100万升献血者的血液。本文将阐述如何用新鲜冰冻胎盘制备高质量的白蛋白。胎盘白蛋白的制备 1.提取胎盘血:冰冻胎盘在低浓度乙醇/盐溶液中快速融化,并在低温中挤压。 2.乙醇法分离血浆蛋白:分四步。第1步:25%乙醇、中性pH,分离球蛋白;第2步:25%乙醇加氯仿去除血红蛋白、脂蛋白及类脂;第3步:25%乙醇、酸性pH,去掉残留的α及β球蛋白;第4步:75%乙醇、酸性pH,去除血型物质、多糖体和可能存在的微量核酸及病毒。以上四步中白蛋白一直处于溶解状态,可避免蛋白分子的变性和聚合成多聚体。 3.层析纯化:分三步。第1步用阴离子交换剂浓缩白蛋白并去除乙醇及氯仿,第2、3步使用部分疏水的阴离子交换剂去除色素、各种疏水杂质如类脂以及偶尔存在的热原质。
Mérieux has been using placentas to extract plasma proteins since 1950. Each placenta contains about 200 ml of blood, which means saving 1 million liters of blood from the blood donor each year. This article explains how to prepare high quality albumin from fresh frozen placenta. Preparation of placental albumin 1. Extraction of placental blood: The frozen placenta melts rapidly in a low concentration ethanol / saline solution and is compressed at low temperatures. 2. ethanol separation of plasma protein: divided into four steps. Step 1: 25% ethanol, neutral pH, globulin; Step 2: 25% ethanol plus chloroform to remove hemoglobin, lipoproteins and lipids; Step 3: 25% ethanol, acidic pH, β globulin; Step 4: 75% ethanol, acidic pH, remove blood type substances, polysaccharides and possible trace nucleic acids and viruses. In the above four steps of albumin has been in solution, to avoid the denaturation of protein molecules and polymerization into multimers. 3. Chromatography purification: in three steps. Step 1 Concentrate the albumin with anion exchanger and remove ethanol and chloroform. Steps 2 and 3 use a partially hydrophobic anion exchanger to remove pigments, various hydrophobic impurities such as lipids and occasional pyrogens.