【摘要】 本文研究了人白细胞干扰素的制备与纯化技术,并作了某些改进。①诱生干扰素方面,用去酚红Earle液低速离心去除剩余红细胞;用NDV-F株代替仙台病毒作诱生剂,粗制干扰素滴度平均达15,000单位/毫升。②KCNS-乙醇法纯化干扰素方面,省略粗制干扰素的酸处理步骤,乙醇溶解KCNS沉淀的干扰素时pH值控制在3.5～4.5,然后用pH5.0,5.6的酸性乙醇分级去除杂蛋白。本法可使干扰素提纯约600倍,比活性10~6位/毫克蛋白,回收率62％。本法也适用人脐血干扰素的纯化。更多还原

【Abstract】 In the present paper the procedure for preparation and purification of human leukocyte interferons were studied and the following improvements were suggested. Phenol red free Earle’s solution was used to remove residual red cells after treatment of buffy coat with 0.83% NH4Cl F strain of Newcastle disease virus was adopted as interferon inducer instead of Sendai virus. The titer of crude interferon obtained was about 15,000 units per ml. As regard to the KCNS-ethanol purification procedure, the pH value of ethanol must be controlled between 3.5~4.5 to dissolve KCNS-precipitated interferons and at 5.0, 5.6 to remove contaminated proteins in interferon preparation. Treatment of crude IF at pH 2.0 was omitted. Using the method mentioned above, human leukocyte IFs could be purified 600 times. The specific activity of purified interferons was about 106/mg protein with 62% revovery. Interferons preparated from human umbilical cord blood could also be purified by this method.This paper offers a method for large-scale production of high titer purified human leukocyte interferons for clinical trial.更多还原