Schlaff S
Endocrinology. 1976 Feb;98(2):527-33. doi: 10.1210/endo-98-2-527.
A specific and sensitive assay for recombination of the subunits of hCG and other glycoprotein hormones (LH, FSH, and TSH) is presented. The assay depends on the ability of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) to separate clearly the subunits from native hormone. Iodinated subunit is incubated with its identical unlabeled subunit and complementary subunit. Quantitative displacement of the radioactivity to the native hormone position as followed by gel slicing is used as a direct index of recombination. The assay is reproducible, sensitive to less than one picomole of hormone formed, and offers a simple means of evaluating recombination of glycoprotein chains as distinct from biological activity.
本文介绍了一种用于检测人绒毛膜促性腺激素(hCG)及其他糖蛋白激素(促黄体生成素(LH)、促卵泡生成素(FSH)和促甲状腺激素(TSH))亚基重组的特异性灵敏检测方法。该检测方法基于十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)能够清晰分离天然激素亚基的能力。将碘化亚基与相同的未标记亚基及互补亚基一起孵育。通过凝胶切片追踪放射性向天然激素位置的定量转移,以此作为重组的直接指标。该检测方法具有可重复性,对形成的激素检测灵敏度低于1皮摩尔,并且提供了一种评估糖蛋白链重组的简单方法,与生物活性不同。