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利用白蛋白磁性微粒的免疫和非免疫T4放射分析

Immune and non-immune T4 radioassays utilizing albumin magnetic microparticles.

作者信息

Ithakissios D S, Kubiatowicz D O, Windorski D C, Wicks J H

出版信息

Clin Chim Acta. 1978 Mar 1;84(1-2):69-84. doi: 10.1016/0009-8981(78)90478-3.

Abstract

Albumin magnetic microparticles reversibly adsorb thyroxine. They quickly establish equilibrium allowing time and temperature independent measurements in T4 radioassays. We used these particles to compare the efficiency of NaOH, HCl, pepsin, sodium trichloroacetate, and 8-anilino-1-naphthalene sulfonic acid to release 125I-T4 from serum, T4-free serum and human serum albumin. We found that the efficiency of the reagents to extract 125I-T4 depended on the concentration and type of proteins to which the labelled hormone was bound. Pepsin was the most effective reagent and we utilized it for a T4 radioimmunoassay, in which albumin magnetic microparticles were used to separate free from bound hormone. We also utilized the particles in a T4 non-immune radioassay. Both assays accurately measured total serum T4, however the radioimmunoassay was simpler, less dependent on protein content of serum, required a smaller serum sample and provided slightly higher T4 values. We describe a magnetic rack which allows simultaneous handling of fifty individual tubes with an intra-assay C.V. of 2.1% for the radioimmunoassay and 2.3% for the non-immune assay and an inter-assay C.V. of 3.1%, respectively.

摘要

白蛋白磁性微粒可可逆地吸附甲状腺素。它们能迅速建立平衡,使得在T4放射免疫测定中进行与时间和温度无关的测量成为可能。我们使用这些微粒比较了氢氧化钠、盐酸、胃蛋白酶、三氯乙酸钠和8-苯胺基-1-萘磺酸从血清、无T4血清和人血清白蛋白中释放125I-T4的效率。我们发现,试剂提取125I-T4的效率取决于标记激素所结合的蛋白质的浓度和类型。胃蛋白酶是最有效的试剂,我们将其用于T4放射免疫测定,其中使用白蛋白磁性微粒分离游离激素和结合激素。我们还将这些微粒用于T4非免疫放射测定。两种测定方法均能准确测量血清总T4,然而放射免疫测定更简单,对血清蛋白质含量的依赖性更小,所需血清样本量更小,且提供的T4值略高。我们描述了一种磁性架,它可以同时处理50个单独的试管,放射免疫测定的批内变异系数为2.1%,非免疫测定的批内变异系数为2.3%,批间变异系数分别为3.1%。

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