Kawakita M, Cabrer B, Taira H, Rebello M, Slattery E, Weideli H, Lengyel P
J Biol Chem. 1978 Jan 25;253(2):598-602.
Interferon production was induced in mouse Ehrlich ascites tumor cells by infection with Newcastle disease virus. The interferon produced was purified by precipitation with ammonium sulfate, chromatography on carboxymethyl-Sephadex, treatment with blue dextran and polyethylene glycol, gel filtration on Bio-Gel P-60 and Bio-Gel P-200, chromatography on phosphocellulose, isoelectric focusing, and chromatography on octyl-Sepharose. The specific activity of the product was 1.6 x 10(9) NIH mouse interferon reference standard units/mg of protein. Electrophoresis in polyacrylamide gels in the presence of sodium dodecyl sulfate indicated that the apparent molecular weight of the interferon-active material ranged from 25,000 to 35,000. As revealed by staining the gels with Coomassie brilliant blue, the interferon activity co-migrated with the major, broad protein band. Minor, stainable bands of proteins were free of interferon activity and their apparent molecular weight was smaller than 12,000.
用新城疫病毒感染小鼠艾氏腹水瘤细胞可诱导干扰素产生。产生的干扰素通过硫酸铵沉淀、羧甲基葡聚糖凝胶柱层析、蓝色葡聚糖和聚乙二醇处理、Bio-Gel P-60和Bio-Gel P-200凝胶过滤、磷酸纤维素柱层析、等电聚焦以及辛基葡聚糖凝胶柱层析进行纯化。产物的比活性为1.6×10⁹ NIH小鼠干扰素参考标准单位/毫克蛋白质。在十二烷基硫酸钠存在下进行的聚丙烯酰胺凝胶电泳表明，干扰素活性物质的表观分子量在25,000至35,000之间。用考马斯亮蓝对凝胶染色显示，干扰素活性与主要的宽蛋白带共迁移。次要的、可染色的蛋白带没有干扰素活性，其表观分子量小于12,000。
