【作者】 徐兵； 潘晨； 蒋晓织； 林曼芝； 陈敏； 林灼； 陈怡； 陈惠聪；

【Author】 XU Bing PAN Chen JIANG Xiao-zhi.Fuzhou Hospital of Infectious Disease,Fuzhou 350025,China

【机构】 福州市传染病医院肝病研究所；

【摘要】 目的:有选择地快速回收人工肝置换下来的病人血浆中多种蛋白,以期用于自体回输治疗。方法:我们设计了一种梯度 pH 共沉淀法(从 pH7.3开始,逐滴加入1mol/L 柠檬酸溶液,5h 加完,使溶液 pH 值呈渐进梯度变化,至终点 pH5.1),用此法配合盐析法(NaCl 饱和度33%,4℃反应5.5h;)或低温乙醇沉淀法(乙醇浓度40%,-7℃反应5.5h),使等电点在 pH7.3～5.1范围内的各蛋白连环碰撞聚集而析出,形成多蛋白共沉淀,再一併回收,弃上清液以清除毒物。结果:在 pH7.3～5.1范围内,本文用梯度 pH 低温乙醇法和梯度 pH 盐析法分别回收了41%(25/61 g/L)和50%(29/57 g/L)的血浆蛋白,上清液中未回收的蛋白主要是白蛋白(及所结合的胆红素)。两法回收的蛋白溶液中,总胆红素分别下降了13倍(低温乙醇法) 和16倍(盐析法),HBV DNA 降为阴性(PCR 定量)。结论:在高浓度中性盐或低温乙醇条件下,渐进微调溶液 pH 值,可使一连串有相邻等电点的蛋白共沉淀,由此可用本文的梯度 pH 共沉淀法选择性快速回收人工肝丢弃血浆中总蛋白以便用于自体回输治疗。更多还原

【Abstract】 Objective To recover selectively the proteins in plasma displaced out in the artificial-liver treatment for the purpose of self-retransfusion of treatment.Methods With the low-temperature ethanol precipitation（ethanol 40%,-7℃,5.5 h） or the salting out （NaCl,degree of saturation 33%,4℃,5.5 h） and by adding 1mol/L C₆H₈O₇H₂O slowly in the plasma solution （changing gradually the isoelectric point pH values from7.3 to 5.1）,we recovered the selective total proteins in the plasma （range from pH 5.1 to 7.3） and discarded the supemants including the bilirubin,albumin and other toxicants.Results In the range of pH 5.1～7.3,50% of the total plasma proteins has been recovered by the gradient pH low-temperature ethanol co-precipitation and 41% by the gradient pH salting out,both methods discarded most albumin in the supernants.The levels of total bilirubin decreased by 16 times and 13 times for the two methods respectively and the levels of HBV DNA decreased to negative for both.Conclusion Under the condition of high degree of concentration of NaCl or low-temperature ethanol,and changing gradually the pH values,the proteins of near-by isoelectric points could be precipitated together（co-precipitation）.By the methods above,with centrifuge seperation only once,we recovered the selective total proteins in the plasma and decreased the levels of bilirubin significantly,and the self-proteins recovered might be used for self-retransfusion in future.更多还原