Iwakura Y, Yonehara S, Kawade Y
J Biol Chem. 1978 Jul 25;253(14):5074-9.
Mouse L cell interferon induced by Newcastle disease virus was purified by a combination of salt precipitation, ion exchange chromatography, gel filtration, and affinity chromatography using anti-interferon anti-body. The preparation was labeled with 125I at a later step of purification, which served as an index of protein concentration. Two main species of interferon molecules differing in molecular weight were separated from each other, and the final preparations obtained were shown to be essentially pure by polyacrylamide gel electrophoresis in the absence as well as in the presence of sodium dodecyl sulfate. The highest specific activities obtained were 2.6 X 10(9) and 7.3 X 10(8) international units/mg of protein (determined by 125I radioactivity) for the 40,000- and 24,000-dalton species, respectively. The preparations at different steps of purification, including those of the highest purity, were tested for the anti-cell growth activity. Their activities were found to be indistinguishable from each other when compared at the same antiviral doses, indicating that the anti-cell growth activity is carried by the interferon molecules themselves.
用盐析、离子交换层析、凝胶过滤以及使用抗干扰素抗体的亲和层析相结合的方法,纯化了新城疫病毒诱导产生的小鼠L细胞干扰素。在纯化的后期步骤中,用125I对该制剂进行标记,以此作为蛋白质浓度的指标。分离出了分子量不同的两种主要干扰素分子,通过在有无十二烷基硫酸钠存在的情况下进行聚丙烯酰胺凝胶电泳,结果表明最终获得的制剂基本纯净。对于分子量为40,000和24,000道尔顿的两种干扰素,所获得的最高比活性分别为2.6×10(9)和7.3×10(8)国际单位/毫克蛋白质(通过125I放射性测定)。对纯化不同阶段的制剂,包括纯度最高的制剂,进行了抗细胞生长活性测试。当在相同抗病毒剂量下进行比较时,发现它们的活性彼此无差异,这表明抗细胞生长活性由干扰素分子本身携带。