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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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至尊木蟲 (職業(yè)作家)
| 對(duì)EP進(jìn)行純化的前期工作采用貧血羊血漿,因?yàn)榭梢缘玫较鄬?duì)大量而尿中激素的存在是不確定的。這種血漿具有每毫克蛋白0.007單位的效價(jià),經(jīng)過(guò)廣泛的序列凈化程序,可以分離出效價(jià)為8250單位/毫克的EP(5)。然而,最終產(chǎn)率僅為0.4%。盡管從血漿中提純的EP是常規(guī)操作,可以明顯看出貧血患者尿提供了污染蛋白很少激素來(lái)源。其回收率為18.8%,或超過(guò)血漿Ep收率的50倍以上。Miyake,Kung 和Goldwasser(7) 將他們?yōu)閮艋蜓獫{ EP開發(fā)的方法應(yīng)用到人尿制備EP。通過(guò)DEAE -纖維素,苯酚-對(duì)氨基水楊酸,乙醇沉淀,DEAE -瓊脂糖,磺酸-葡聚糖,凝膠滲透和羥基磷灰石色譜相結(jié)合的組合處理,他們獲得了70400單位/毫克蛋白效價(jià)的EP, 產(chǎn)率為21 %。 |
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