液相色谱法纯化人血清C2蛋白

Purification of C2 from human serum by liquid chromatography

目的 建立一种从血清中提纯 C2的方法 ,得到稳定的、有活性的纯品 ,便于今后进一步研究其生理病理功能 .方法 利用冷球蛋白沉淀、优球蛋白沉淀、离子交换色谱、硫酸铵沉淀和亲和色谱几种方法 ,将血清中不同性质的杂蛋白除去 ,选择相对简单且分离效果好的条件 ,提高终产物中目标蛋白 C2的纯度和含量 .结果 成功地从 1L血浆中提纯到了 2 .49m g C2 ,产率为 12 .46 % ,活性回收率为 13.6 8% .终产物在 p H中性时稳定 ,且无需加入蛋白稳定剂 .结论 本法与以往纯化 C2的方法相比 ,简便可行 ,产率及活性回收率较高 。

AIM To establish a method for purification of C2 from human serum, to obtain the stable and active purification and to study its physiological and pathological function easily in the future. METHODS Using a combination of cryoglobulin precipitation,euglobulin precipitation, ion exchange chromatography, (NH 4) 2SO 4 precipitation and affinity chromatography, the impurity protein of different quality was removed, and the easy and effective condition was selected to increase the purity and content of C2 in the final product. RESULTS 2 49 mg final product was homogeneous by the criterion of SDS PAGE representing the 12.46% yield and 13.68% activity recovery from 1000 mL serum. They were stable in pH neutral and no stabilizing agent was needed. CONCLUSION Compared with the past method, this method with high yield and activity can be used for laboratory research and large scale preparation.