Allen G K, Rosenquist B D
Department of Veterinary Microbiology, College of Veterinary Medicine, University of Missouri, Columbia 65211.
Am J Vet Res. 1988 Jun;49(6):762-5.
Bovine fibroblast interferon (BoF-IFN), produced in bovine embryonic kidney cell cultures by priming and infection with bluetongue virus, was partially purified by controlled pore glass chromatography. The partially purified BoF-IFN then was subjected to beaded agarose affinity chromatography in 2 distinct fractions--1 after the addition of 1M NaCl and the other one after the addition of 1.5M NaCl containing 50% ethylene glycol. Analysis of fractions by sodium dodecyl sulfate/polyacrylamide-gel electrophoresis revealed a broad molecular weight range (14,900 to 27,900) for IFN eluted by 1M NaCl, and 2 discrete molecular weight ranges (16,000 to 19,500 and 28,300 to 34,000) for IFN eluted by 1.5M NaCl containing 50% ethylene glycol. The specific activity of the IFN eluted with 1.5M NaCl containing ethylene glycol was 2.85 X 10(6) U/mg of protein, compared with 5.7 X 10(5) U/mg of protein in the controlled pore glass-purified IFN.
通过用蓝舌病毒引发和感染牛胚胎肾细胞培养物产生的牛成纤维细胞干扰素（BoF-IFN），经可控孔径玻璃色谱法进行部分纯化。然后，将部分纯化的BoF-IFN进行琼脂糖珠亲和色谱，分为两个不同的组分——一个在加入1M NaCl后得到，另一个在加入含50%乙二醇的1.5M NaCl后得到。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳对各组分进行分析，结果显示，用1M NaCl洗脱的干扰素分子量范围较宽（14,900至27,900），而用含50%乙二醇的1.5M NaCl洗脱的干扰素分子量范围有两个离散区间（16,000至19,500和28,300至34,000）。与经可控孔径玻璃纯化的干扰素中每毫克蛋白质含5.7×10⁵ 单位相比，用含乙二醇的1.5M NaCl洗脱的干扰素的比活性为每毫克蛋白质2.85×10⁶ 单位。
