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叙利亚仓鼠干扰素的部分纯化及特性分析

Partial purification and characterization of Syrian hamster interferon.

作者信息

Bollin E, Sulkowski E

出版信息

J Gen Virol. 1981 Feb;52(Pt 2):227-33. doi: 10.1099/0022-1317-52-2-227.

Abstract

Syrian golden hamster interferon was made by stimulating secondary or benzo(a)pyrene-transformed embryo cells with Newcastle disease virus. Titres of 1000 to 6000 units/ml of tissue culture fluid were obtained. The ionic properties of this interferon were characterized by chromatography on cation and anion exchangers and also by isoelectric focusing when two components were observed: a major component, pI 5.8 and a minor component, pI 6.6. Hamster interferon was partially purified on a tandem of sorbents of diverse chromatographic bias: anion exchanger lead to metal chelate lead to hydrophobic ligand. The purified preparation had a specific activity of about 1 X 10(6) units/mg protein; the recovery of activity was nearly complete. The apparent mol. wt. of hamster interferon, as estimated by SDS-polyacrylamide gel electrophoresis under non-reducing conditions, is 17000. Crude Syrian hamster interferon preparations had some activity on mouse cells; this heterologous activity was entirely due to a small subpopulation of interferon molecules which could be isolated on phenylagarose.

摘要

叙利亚金黄地鼠干扰素是通过用新城疫病毒刺激传代或经苯并(a)芘转化的胚胎细胞制备而成。组织培养液的效价达到了1000至6000单位/毫升。该干扰素的离子特性通过在阳离子和阴离子交换剂上的色谱分析以及等电聚焦来表征,观察到有两个组分:一个主要组分,pI 5.8,和一个次要组分,pI 6.6。地鼠干扰素在一系列具有不同色谱偏向的吸附剂上进行了部分纯化:阴离子交换剂接金属螯合剂接疏水配体。纯化后的制剂比活性约为1×10(6)单位/毫克蛋白质;活性回收率几乎达到100%。在非还原条件下通过SDS-聚丙烯酰胺凝胶电泳估计,地鼠干扰素的表观分子量为17000。叙利亚地鼠干扰素粗制品对小鼠细胞有一定活性;这种异源活性完全归因于一小部分可在苯琼脂糖上分离的干扰素分子。

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