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一种用于检测人皮质类固醇结合球蛋白的固相放射免疫测定法。

A solid-phase radioimmunoassay for human corticosteroid binding globulin.

作者信息

Robinson P A, Langley M S, Hammond G L

出版信息

J Endocrinol. 1985 Feb;104(2):259-67. doi: 10.1677/joe.0.1040259.

Abstract

A radioimmunoassay (RIA) for human corticosteroid binding globulin (CBG) has been developed using 125I-labelled CBG and a monospecific solid-phase CBG-antiserum (CBG-Ab-cellulose). In an RIA of serum CBG concentrations, pure CBG standards (1-100 ng protein) or samples (1:200) were incubated (16 h at 20 degrees C) with 125I-labelled CBG and CBG-Ab-cellulose. After addition of 2 ml 0.9% NaCl, the tubes were centrifuged, supernatants were aspirated and the 125I-labelled CBG bound to the CBG-Ab-cellulose pellet was counted. The specificity of the RIA was confirmed by parallel displacement curves for serial dilutions of male, female and pregnancy sera, as well as pure CBG standards. The mean +/- S.D. recovery (99 +/- 8%) of pure CBG (1.6-25.0 ng) added to a diluted serum sample verified the accuracy of the method, and a good correlation (r = 0.97; n = 43) existed between serum CBG cortisol binding capacity (nmol/l) measurements and CBG concentrations (mg protein/l) measured by RIA. Intra- and interassay precisions (C.V.) at low to high serum CBG concentrations were less than 5% and less than 9% respectively. The mean +/- S.D. serum CBG concentrations (mg protein/l) measured by the RIA were: 21.8 +/- 4.6 in boys (n = 12), 20.0 +/- 4.2 in girls (n = 9), 20.7 +/- 2.7 in men (n = 6), 20.5 +/- 2.9 in women (n = 6) and 47.1 +/- 10.5 in pregnant women (n = 5).(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

已开发出一种用于检测人皮质类固醇结合球蛋白(CBG)的放射免疫分析方法(RIA),该方法使用125I标记的CBG和单特异性固相CBG抗血清(CBG-抗体纤维素)。在血清CBG浓度的RIA检测中,将纯CBG标准品(1 - 100 ng蛋白质)或样品(1:200)与125I标记的CBG和CBG-抗体纤维素一起孵育(20℃下16小时)。加入2 ml 0.9%氯化钠后,将试管离心,吸出上清液,对结合在CBG-抗体纤维素沉淀上的125I标记的CBG进行计数。通过对男性、女性和妊娠血清以及纯CBG标准品的系列稀释液的平行置换曲线,证实了该RIA的特异性。向稀释的血清样品中添加纯CBG(1.6 - 25.0 ng)后的平均回收率(99±8%)验证了该方法的准确性,并且血清CBG皮质醇结合能力(nmol/l)测量值与通过RIA测量的CBG浓度(mg蛋白质/l)之间存在良好的相关性(r = 0.97;n = 43)。在低至高血清CBG浓度下,批内和批间精密度(变异系数)分别小于5%和小于9%。通过RIA测量的平均血清CBG浓度(mg蛋白质/l)为:男孩(n = 12)为21.8±4.6,女孩(n = 9)为20.0±4.2,男性(n = 6)为20.7±2.7,女性(n = 6)为20.5±2.9,孕妇(n = 5)为47.1±10.5。(摘要截短于250字)

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