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人血清甲状腺球蛋白结合能力放射免疫测定的进一步观察

Further observations on radioimmunoassay for thyroglobulin-binding capacity in human serum.

作者信息

Salabè G B

出版信息

Ric Clin Lab. 1980 Jul-Sep;10(3):533-44. doi: 10.1007/BF02938799.

Abstract

A simplified radioimmunoassay for measuring thyroglobulin-binding capacity in human serum is described. Thyroglobulin (Tg) is labelled with lactoperoxidase (specific activity 5 microCi/microgram) which makes it possible to measure a binding capacity of more than 10 ng/ml of serum. The standard curve is plotted measuring the percentage of bound 125I-Tg against the dilution of 1 mg/ml Tg capacity from a pool of Tg antibodies from different individual serum samples. When polyethyleneglycol and a second antibody were compared in their precipitation of the immune complexes, the latter provided a better precipitation. The stability of the labelled Tg stored at --20 degrees C was checked at different intervals of time by ultracentrifugation and immunoreactivity. It was found that the labelled Tg can be used up to one month after iodination. Fragments produced during storage at --20 degrees C only partially reacted with Tg antibodies. Screening of 300 serum samples was performed by haemagglutination and binding capacity; of 45 positive serum samples, 28 were positive only by binding capacity, thus demonstrating that this method is far more sensitive, accurate and precise than those currently employed.

摘要

本文描述了一种用于测量人血清中甲状腺球蛋白结合能力的简化放射免疫分析方法。甲状腺球蛋白(Tg)用乳过氧化物酶(比活为5微居里/微克)进行标记,这使得能够测量血清中超过10纳克/毫升的结合能力。通过测量结合的125I-Tg的百分比与来自不同个体血清样本的Tg抗体池的1毫克/毫升Tg能力的稀释度来绘制标准曲线。当比较聚乙二醇和第二抗体对免疫复合物的沉淀作用时,后者沉淀效果更好。通过超速离心和免疫反应性在不同时间间隔检查在-20℃储存的标记Tg的稳定性。发现标记的Tg在碘化后最多可使用一个月。在-20℃储存期间产生的片段仅与Tg抗体部分反应。通过血凝和结合能力对300份血清样本进行筛选;在45份阳性血清样本中,28份仅通过结合能力呈阳性,因此表明该方法比目前使用的方法更灵敏、准确和精确。

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