Sawatzki G, Anselstetter V, Kubanek B
Biochim Biophys Acta. 1981 Jan 30;667(1):132-8. doi: 10.1016/0005-2795(81)90073-8.
A new method for the isolation of considerable quantities of mouse transferrin is described. This technique employs salting-out chromatography on Sepharose CL-6B, a new step in the preparation of plasma proteins. This step is followed by ion-exchange chromatography on DEAE-Sepharose CL-6B and gel filtration on Sephacryl S-200. The isolated mouse transferrin was shown to be pure by immunoelectrophoresis, sodium dodecyl sulphate-polyacrylamide gel electrophoresis and by the 465 nm/410 nm ratio of absorbances being 1.41. The molecular weight was determined to be about 77 500. The advantages of this procedure are that it is reproducible, gives a high recovery, and can be extended to a larger scale. The advantage over other protein purification techniques is its general utility, due to the fact that there is no need for species-specific antibodies. The application of this method offers a rapid purification of sufficient quantities of mouse transferrin essential for the elucidation of biological functions of this protein and investigations of its molecular structure.
本文描述了一种分离大量小鼠转铁蛋白的新方法。该技术采用琼脂糖凝胶CL-6B盐析色谱法，这是血浆蛋白制备中的一个新步骤。接下来是在DEAE-琼脂糖凝胶CL-6B上进行离子交换色谱法以及在Sephacryl S-200上进行凝胶过滤。通过免疫电泳、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及465nm/410nm吸光度比值为1.41，证明分离出的小鼠转铁蛋白是纯的。其分子量测定约为77500。该方法的优点是可重复、回收率高且可扩大规模。与其他蛋白质纯化技术相比，其优势在于通用性强，因为无需种属特异性抗体。该方法的应用为快速纯化足够量的小鼠转铁蛋白提供了可能，这对于阐明该蛋白的生物学功能及其分子结构研究至关重要。
