Pekary A E, Hershman J M, Parlow A F
J Clin Endocrinol Metab. 1975 Oct;41(4):676-84. doi: 10.1210/jcem-41-4-676.
A high-sensitivity radioimmunoassay for human thyroid-stimulating hormone (hTSH) has been developed utilizing a highly specific rabbit antibody (developed by AFP) to hTSH with an affinity constant of 4.4 x 10(11) M-1. Optimal conditions which minimized both 125I-hTSH radiation damage and 125I dissociation included storage at -60 C in 0.25% albumin. Repetitive freezing and thawing did not cause loss of structural integrity or of full immunochemical reactivity of 125I-hTSH. Effects of human and calf sera on the precipitation of first antibody-bound 125I-hTSH by second antibody have been tabulated; the addition of TSH-free human serum to the standard curve is necessary to compensate for the serum effect. The minimum detectable amount of TSH in the new assay at a 1:1,000,000 final dilution of the hTSH antibody is 0.02 muU/tube. The mean normal hTSH value of 1.5 +/- 1.0 muU/ml (mean +/- SD) fell within the central one-third of the logit (B/Bo) plot where antigen concentrations are measured with highest precision. This assay gives good precision and reproducibility of measurements throughout the normal range of serum TSH concentrations.
利用一种对人促甲状腺激素（hTSH）具有高度特异性的兔抗体（由甲胎蛋白研发），开发了一种高灵敏度放射免疫分析法来检测人促甲状腺激素，该抗体的亲和常数为4.4×10¹¹M⁻¹。将¹²⁵I-hTSH的辐射损伤和¹²⁵I解离降至最低的最佳条件包括在-60℃下于0.25%白蛋白中储存。反复冻融不会导致¹²⁵I-hTSH的结构完整性丧失或完全免疫化学反应性丧失。已将人和小牛血清对第二抗体沉淀第一抗体结合的¹²⁵I-hTSH的影响制成表格；向标准曲线中加入无TSH的人血清对于补偿血清效应是必要的。在hTSH抗体最终稀释至1:1,000,000时，新分析方法中TSH的最低检测量为0.02μU/管。正常hTSH的平均数值为1.5±1.0μU/ml（平均值±标准差），落在对数概率（B/Bo）图的中间三分之一范围内，在此范围内抗原浓度的测量精度最高。该分析方法在血清TSH浓度的整个正常范围内具有良好的测量精度和可重复性。
