Motoyoshi K, Takaku F, Miura Y
Exp Hematol. 1983 May;11(5):389-93.
Marked loss of colony stimulating activity was noted when the highly purified human urinary colony stimulating factor (CSFHU-5) dissolved in 0.1 M Tris-HCl buffer (pH 7) was passed through ethylene oxide sterilized Millipore filters. The addition of 0.5 mg/ml polyethylene glycol (PEG) or 10 mg/ml bovine serum albumin (BSA) to the CSFHU solutions resulted in almost complete protection against the CSF loss. A less pure fraction (CSFHU-2) did not lose activity upon filtration. More than 90% of the 125I-CSFHU dissolved in 0.1 M Tris-HCl buffer (pH 7) bound to the ethylene oxide sterilized Millipore filters. The addition of PEG or BSA to the 125I-CSFHU solutions resulted in almost complete prevention of 125I-CSFHU binding to the membranes. These findings indicate that Millipore membranes can bind significant quantities of the highly purified CSFHU, and the addition of PEG to all buffer systems is a useful way to prevent the CSF loss throughout the purification.
当溶解于0.1 M Tris-HCl缓冲液(pH 7)中的高度纯化的人尿集落刺激因子(CSFHU-5)通过环氧乙烷灭菌的密理博滤器时,发现集落刺激活性显著丧失。向CSFHU溶液中添加0.5 mg/ml聚乙二醇(PEG)或10 mg/ml牛血清白蛋白(BSA)可几乎完全防止CSF活性丧失。纯度较低的组分(CSFHU-2)在过滤时不会丧失活性。溶解于0.1 M Tris-HCl缓冲液(pH 7)中的125I-CSFHU超过90%与环氧乙烷灭菌的密理博滤器结合。向125I-CSFHU溶液中添加PEG或BSA可几乎完全防止125I-CSFHU与膜结合。这些发现表明,密理博膜可结合大量高度纯化的CSFHU,并且在所有缓冲系统中添加PEG是防止整个纯化过程中CSF活性丧失的一种有效方法。