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| The initial work on purification of Ep was performed with plasma from anemic sheep,since relatively large volumes could be obtained and the presence of the hormone in urine was uncertain .Such plasma had a potency of 0.007 units/mg of protein ,and after an extensive sequence of purification procedures, Ep with a protency of 8250 units/mg was isolated(5).however,the final yield was only 0.4%.While the purification of Ep from plasma was proceeding ,it became evident that urine of anemic patients provided a source of hormone with much less contaminatiing protein. Their recovery was 18.8%,or more than 50 times greater than that with plasma Ep.Miyake, Kung&Goldwasser(7) have since applied the methodology they developed for purifying sheep palsma Ep to human urinary preparations. By a combination of treatment with DEAE-cellulose ,phenol-p-aminosalicylate , ethanol precipitation, DEAE-agarose, sulfopropyl-Sephadex,gel permeation, and hydroxylapatite chromatography, they obtained Ep at a potency of 70,400 units/mg of protein with a 21%yield. |
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wjmsicuan(金币+10, 翻译EPI+1): 翻译的很认真 2011-03-05 10:31:43
| 对EP进行纯化的前期工作采用贫血羊血浆，因为可以得到相对大量而尿中激素的存在是不确定的。这种血浆具有每毫克蛋白0.007单位的效价，经过广泛的序列净化程序，可以分离出效价为8250单位/毫克的EP（5）。然而，最终产率仅为0.4％。尽管从血浆中提纯的EP是常规操作，可以明显看出贫血患者尿提供了污染蛋白很少激素来源。其回收率为18.8％，或超过血浆Ep收率的50倍以上。Miyake，Kung 和Goldwasser(7) 将他们为净化羊血浆 EP开发的方法应用到人尿制备EP。通过DEAE -纤维素，苯酚-对氨基水杨酸，乙醇沉淀，DEAE -琼脂糖，磺酸-葡聚糖，凝胶渗透和羟基磷灰石色谱相结合的组合处理，他们获得了70400单位/毫克蛋白效价的EP， 产率为21 ％。 |
2楼2011-03-04 08:31:32