Wiley B B, Glasgow L A, Rogolsky M
Infect Immun. 1976 Feb;13(2):513-20. doi: 10.1128/iai.13.2.513-520.1976.
Exfoliative toxin (ET) from a phage group II Staphylococcus aureus strain causing staphylococcal scalded-skin syndrome was purified by electrofocusing. Ampholytes and salts were removed from the final product by column chromatography on G-50 Sephadex. Sodium dodecyl sulfate-polyacrylamide gels of the final product yielded a single band upon gel electrophoresis, even when 60 mug of protein was placed in the gels. Radiolabeling of the purified toxin with 125I yielded a product that still caused exfoliation of suckling mice, indicating that the toxin was still biologically active. A radioimmunobinding assay was developed and used to test rabbit and human sera for antibodies to exfoliative toxin. Although the maximum percentage of binding was not as high as expected (approximately 40%), it was postulated that either iodination had not been sufficiently vigorous or the toxin had sustained immunological damage. The assay was reproducible and more sensitive than the existing neutralization method and readily applicable to the testing of human sera for exfoliative toxin antibodies.
通过电聚焦法对引起葡萄球菌烫伤样皮肤综合征的噬菌体Ⅱ组金黄色葡萄球菌菌株产生的剥脱毒素（ET）进行了纯化。通过Sephadex G - 50柱色谱从最终产物中去除两性电解质和盐。最终产物的十二烷基硫酸钠 - 聚丙烯酰胺凝胶在凝胶电泳时产生单一条带，即使在凝胶中加入60微克蛋白质时也是如此。用125I对纯化毒素进行放射性标记产生的产物仍能引起乳鼠脱皮，表明该毒素仍具有生物活性。开发了一种放射免疫结合试验并用于检测兔和人血清中针对剥脱毒素的抗体。尽管最大结合百分比不如预期的高（约40%），但推测要么碘化不够充分，要么毒素受到了免疫损伤。该试验具有可重复性，比现有的中和方法更灵敏，并且易于应用于检测人血清中的剥脱毒素抗体。
