用于组织培养的人促红细胞生成素的制备。
Preparation of human erythropoietin for tissue culture.
作者信息
Cutler R L, Johnson G R, Nicola N A
出版信息
Exp Hematol. 1985 Sep;13(8):796-801.
A simple two-step procedure has been developed for the production of human urinary erythropoietin suitable for tissue culture use. Lyophilized human urinary protein was fractionated by phenyl-Sepharose chromatography, the erythropoietin binding in 2 M lithium chloride and being eluted by an ascending linear gradient to 6 M guanidine hydrochloride. The erythropoietin was further purified by ethanol (75% vol/vol) precipitation. This two-step procedure resulted in a 25-fold purification of erythropoietin with a 50% recovery of the starting activity. The erythropoietin preparation was suitable for use in murine or human cultures, being free of protease activity, inhibitory factors, colony-stimulating factors, and erythroid-enhancing activities.
已开发出一种简单的两步法来生产适用于组织培养的人尿促红细胞生成素。冻干的人尿蛋白通过苯基琼脂糖色谱法进行分离，促红细胞生成素结合在2M氯化锂中，然后通过升至6M盐酸胍的线性梯度洗脱。促红细胞生成素通过乙醇（75%体积/体积）沉淀进一步纯化。此两步法使促红细胞生成素得到了25倍的纯化，起始活性回收率为50%。该促红细胞生成素制剂适用于小鼠或人类培养，不含蛋白酶活性、抑制因子、集落刺激因子和红细胞生成增强活性。
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