Berthold W, Tan C, Tan Y H
J Biol Chem. 1978 Jul 25;253(14):5206-12.
Interferon was produced from a clonal human fibroblastoid line. This cell line was derived from an established fibroblastoid culture treated with ethylmethane sulfonate and is capable of producing higher amounts human interferon than primary human fibroblast cultures. The interferon produced from this cell line was purified by concanavalin A and phenyl-Sepharose column chromatography followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified interferon was then labeled with [125I]-iodine, with [3H]5-dimethylaminonaphthalene-1-sulfonyl-chloride and with sodium [3H]borohydride after periodate oxidation. An appreciable amount of biological activity was retained after both tritium labelings but not after iodination. When subjected to electrophoresis in the presence of sodium dodecyl sulfate all three radioactive interferon preparations comigrated with the purified interferon preparation as a single protein component with a molecular weight of 19,000. Both purified and radioactive preparations were also shown to migrate with the antiviral activity in polyacrylamide gels in the absence of sodium dodecyl sulfate. The specific activity of purified interferon preparations ranged from 2 X 10(8) to 10 X 10(8) units/mg of protein.
干扰素由一株克隆的人成纤维细胞样细胞系产生。该细胞系源自经甲磺酸乙酯处理的已建立的成纤维细胞样培养物，并且能够产生比原代人成纤维细胞培养物更多的人干扰素。从该细胞系产生的干扰素通过伴刀豆球蛋白A和苯基琼脂糖柱层析，随后进行制备性十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳进行纯化。然后，纯化的干扰素在高碘酸盐氧化后用[125I] - 碘、[3H]5 - 二甲基氨基萘 - 1 - 磺酰氯和[3H]硼氢化钠进行标记。两次氚标记后均保留了相当量的生物活性，但碘化后则没有。当在十二烷基硫酸钠存在下进行电泳时，所有三种放射性干扰素制剂与纯化的干扰素制剂一起作为单一蛋白质组分迁移，其分子量为19,000。在不存在十二烷基硫酸钠的情况下，纯化的和放射性的制剂在聚丙烯酰胺凝胶中也显示出与抗病毒活性一起迁移。纯化的干扰素制剂的比活性范围为2×10(8)至10×10(8)单位/毫克蛋白质。
