荧光激发转移免疫分析法测定人血清中的甲状腺素结合球蛋白（TBG）

Determination of thyroxine binding globulin (TBG) in human serum by fluorescence excitation transfer immunoassay.

作者信息

Van der Werf P, Chang C H

出版信息

J Immunol Methods. 1980;36(3-4):339-47. doi: 10.1016/0022-1759(80)90139-8.

Abstract

Human thyroxine binding globulin (TBG) was purified to near homogeneity by chromatography on thyroxine-substituted Sepharose 4B, DEAE-Sephadex A-50, Sephadex G-150 and n-decylamine-substituted Sepharose 4B. TBG was quantitated by homogeneous fluorescence excitation transfer immunoassay; the preparation of dye-protein conjugates is described. The fluorescence of fluorescein-TBG is quenched when an immune complex is formed with tetramethylrhodamine-antibody. In the presence of unlabeled TBG, the relative fluorescence intensity is increased and can be related quantitatively to the concentration of competing antigen. The described assay for TBG has a range corresponding to serum concentrations of 5--75 microgram/ml; the correlation with radioimmunoassay is 0.96. The coefficients of variation for replicate determinations of single samples of serum with low, normal and elevated TBG concentrations were 3.5%, 2.7% and 2.6% respectively; between-assay variations were 8.8%, 8.6% and 2.6% respectively.

摘要

通过在甲状腺素取代的琼脂糖凝胶4B、二乙氨基乙基葡聚糖A - 50、葡聚糖凝胶G - 150和正癸胺取代的琼脂糖凝胶4B上进行色谱分离，将人甲状腺素结合球蛋白（TBG）纯化至接近均一。采用均相荧光激发转移免疫测定法定量TBG；文中描述了染料 - 蛋白质缀合物的制备方法。当荧光素 - TBG与四甲基罗丹明 - 抗体形成免疫复合物时，其荧光被淬灭。在未标记的TBG存在下，相对荧光强度增加，并且可以与竞争抗原的浓度进行定量关联。所描述的TBG测定法的范围对应于血清浓度5 - 75微克/毫升；与放射免疫测定法的相关性为0.96。对低、正常和升高TBG浓度的血清单一样品进行重复测定的变异系数分别为3.5%、2.7%和2.6%；批间变异分别为8.8%、8.6%和2.6%。