C反应蛋白的纯化
Purification of C-reactive protein.
作者信息
Ikuta T, Okubo H, Yukimura H, Ishibashi H
出版信息
Biochem Int. 1983 Feb;6(2):275-82.
A concise method was designed for preparation of C-reactive protein (CRP) from pleural effusion. By addition of L-alpha-lecithin to the pleural effusion in the presence of calcium ions, a flocculence of the CRP-lecithin complex formed. Subsequent treatment of the CRP-lecithin complex with chloroform and sodium citrate buffer enabled extraction of the CRP in the buffer layer. This extracted CRP was further purified by sequential treatment on column chromatography of DEAE cellulose (DE52) and gel filtration using Sephacryl S-300. The isolated protein was proved to be native CRP with a high degree of purity, as determined by electrophoretical and immunological analysis. The yield was 41.8% recovery from the starting material. E1%(280) of the CRP preparation was estimated to be 18.75.
设计了一种从胸腔积液中制备C反应蛋白（CRP）的简便方法。在钙离子存在的情况下，向胸腔积液中加入L-α-卵磷脂，形成CRP-卵磷脂复合物絮凝物。随后用氯仿和柠檬酸钠缓冲液处理CRP-卵磷脂复合物，使CRP在缓冲层中得以提取。通过在DEAE纤维素（DE52）柱色谱上的连续处理和使用Sephacryl S-300的凝胶过滤，对提取的CRP进行进一步纯化。经电泳和免疫学分析确定，分离得到的蛋白质为高纯度的天然CRP。产率为起始原料回收率的41.8%。CRP制剂的E1%（280）估计为18.75。
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