Biosensor recognition of thyroid-disrupting chemicals using transport proteins
Biosensor recognition of thyroid-disrupting chemicals using transport proteins
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DOI：
10.1021/ac051399i
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发表时间：
2006-02-15
影响因子：
7.4
通讯作者：
Irth, H
中科院分区：
文献类型：
--
作者：
Marchesini, GR;Meulenberg, E;Irth, H
描述了一种新的基于表面等离子体共振的生物传感器分析方法，用于筛选具有甲状腺干扰活性的化学物质的生物效应。甲状腺激素结合球蛋白(Tbg)和重组转甲状腺素(RTtr)两种甲状腺转运蛋白(TPS)在Biacore 3000生物传感器芯片上被应用于甲状腺系统主要激素L-甲状腺激素(T4)的抑制检测。对天然甲状腺激素的测定条件进行了优化，并选择已知的甲状腺干扰物和结构相关的化合物作为模型化合物，在两种测定中测试它们的相对效力(Rp)与T4的比较。所选化合物为卤代酚、卤代双酚、双酚A、3，5-二氯联苯及其羟化代谢物4-羟基-3，5-二氯联苯(4-OHPCB14)。基于TBG的方法对T4具有高度的特异性，而基于rTTR的方法对几种化合物具有较高的敏感性，其中4-羟基PCB14的敏感性最高(Rp=4.4)，其次是四溴双酚A(Rp=1.5)和四氯双酚A(Rp=0.75)。对于已知的与生物效应相关的筛选和可能的新的甲状腺干扰物的鉴定，基于TPS的生物传感器检测比目前使用的放射配基结合分析和免疫沉淀-高效液相色谱检测方法更灵敏(rTTR和基于TBG的检测的IC50分别为13.7+/-1.3和8.6+/-10.7 nm)，操作更容易，替代方法更快(10min/样品)。
Novel surface plasmon resonance-based biosensor assays for the bioeffect-related screening of chemicals with thyroid-disrupting activity are described. Two thyroid transport proteins (TPs), thyroxine binding globulin (TBG) and recombinant transthyretin (rTTR), were applied in an inhibition assay format in a Biacore 3000 using CM5 biosensor chips coated with L-thyroxine (T4), the main hormone of the thyroid system. Assay conditions were optimized for the natural thyroid hormones, and known thyroid disruptors and structurally related compounds were selected as model compounds to be tested in both assays for their relative potency (RP) compared to T4. The chosen compounds were halogenated phenols, halogenated bisphenols, bisphenol A, 3,5-dichlorobiphenyl, and its hydroxylated metabolite 4-hydroxy-3,5-dichlorobiphenyl (4-OH PCB 14). The TBG-based assay was highly specific for T4, and the rTTR-based assay was sensitive toward several compounds, the highest sensitivity (RP = 4.4) being obtained with 4-OH PCB 14, followed by tetrabromobisphenol A (RP = 1.5) and tetrachlorobisphenol A (RP = 0.75). For the bioeffect-related screening of known and identification of possible new thyroid disruptors, the TPs-based biosensor assays were more sensitive (IC50 of 13.7 +/- 1.3 and 8.6 +/- 10.7 nM for the rTTR and the TBG-based assay, respectively), easier to perform, and faster alternatives (10 min/sample) than the currently used methods such as radioligand binding assays and immunoprecipitation-HPLC.