UNIVERSAL ASSAY OF VITELLOGENIN AS A BIOMARKER FOR ENVIRONMENTAL ESTROGENS
UNIVERSAL ASSAY OF VITELLOGENIN AS A BIOMARKER FOR ENVIRONMENTAL ESTROGENS
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DOI：
10.2307/3432500
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发表时间：
1995-10-01
影响因子：
10.4
通讯作者：
SULLIVAN, CV
中科院分区：
文献类型：
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作者：
HEPPELL, SA;DENSLOW, ND;SULLIVAN, CV
卵黄蛋白原（VTG）是卵黄蛋白的前体磷脂糖蛋白，是环境雌激素的理想生物标志物。本研究的目的是开发针对VTG分子保守区域的抗体，这些抗体可以为建立生物测定法以检测任何卵生脊椎动物中的雌激素暴露奠定基础。我们开发了针对纯化虹鳟鱼（虹鳟鱼）VTG产生的单克隆抗体（mAb），并选择了特异性识别从两种遗传学上遥远的脊椎动物，鳟鱼和条纹鲈鱼（岩鲈鱼）纯化的VTG的属性。酶联免疫吸附试验和蛋白质印迹的结果表明，这些单克隆抗体特异性地识别纯化的VTG和VTG或其他雌激素诱导的蛋白质的血浆或血清中的代表性物种的四个脊椎动物类（鱼，两栖动物，爬行动物和鸟类）。产生的所有mAb均为IgM类。针对代表VTG的保守N-末端氨基酸序列的合成共有肽产生多克隆抗血清。Western blotting结果表明，该抗血清能特异性识别不同科硬骨鱼血浆或血清中的VTG。它用于检测来自患有癌症（肝细胞癌和胆管癌）的褐牛头（Ameiurusnebulosus）的血清的Western印迹中的VTG，所述褐黑头（Ameiurusnebulosus）收集自在其正常卵黄发生季节之外的污染的工业场所。我们的研究结果表明，它是可行的，以产生能够识别VTG的抗体，而不考虑物种和开发一个通用的VTG检测是一个可以实现的目标。
Vitellogenin (VTG), the serum phospholipoglycoprotein precursor to egg yolk, is potentially an ideal biomarker for environmental estrogens. This study was undertaken to develop antibodies against conserved regions on the VTG molecule that could form the basis for establishing bioassays to detect estrogen exposure in any oviparous vertebrate. We developed monoclonal antibodies (mAbs) generated against purified rainbow trout (Oncorhynchus mykiss) VTG and selected for the property of specifically recognizing VTG purified from two phylogenetically distant vertebrates, trout and striped bass (Morone saxatilis). Results of enzyme-linked immunosorbent assay and Western blotting indicated that these mAbs specifically recognize purified VTG and VTG or other estrogen-inducible proteins in plasma or serum from representative species of four vertebrate classes (fish, amphibians, reptiles, and birds). Ail of the mAbs generated were IgM class. A polyclonal antiserum was raised against a synthetic consensus peptide representing the conserved N-terminal amino acid sequence of VTG. The results of Western blotting indicate that this antiserum specifically recognizes VTG in plasma or serum from teleost fish of diverse families. it was used to detect VTG in Western blots of serum from brown bullhead (Ameiurus nebulosus) with cancer (hepatocellular and cholangio-carcinoma) collected from a contaminated industrial site outside of their normal vitellogenic season. Our results indicate that it is feasible to generate antibodies capable of recognizing VTG without regard to species and that development of a universal VTG assay is an achievable goal.