Determination of estrogenic activity by LYES-assay (yeast estrogen screen-assay assisted by enzymatic digestion with lyticase)
Determination of estrogenic activity by LYES-assay (yeast estrogen screen-assay assisted by enzymatic digestion with lyticase)
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DOI：
10.1016/j.chemosphere.2004.06.027
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发表时间：
2004-12-01
期刊：
影响因子：
8.8
通讯作者：
Metzger, JW
中科院分区：
文献类型：
--
作者：
Schultis, T;Metzger, JW
为了提高酵母雌激素筛选(YES)法的灵敏度和速度，许多实验室已经建立了测定化合物和环境样品雌激素活性的酵母雌激素筛选(YES)法，我们发展了一种包括酶裂解酶消化步骤的YES法的改进版本。用LYES法测定了天然雌激素(17β-雌二醇E_2和雌酮)、合成雌激素(17α-雌二醇E_2)和药用雌激素(己烯雌酚DES)以及异种雌激素(4-壬基酚NP和5种对羟基苯甲酸酯)的雌激素活性，并与常规的YES法、E-Screen法(MCF-7乳腺肿瘤细胞增殖)和受体结合试验(RB)进行了比较。在E2的情况下，LYES测定法的定量下限(LOQ)明显低于常规的YES测定法。比Rb测定法低两个数量级。与E-Screen法相比，LYES-法的LOQ几乎高出一个数量级。进行LYES检测所需的时间只需7小时，而传统的YES检测需要3至5天。因此，LYES法是一种非常好的替代现有雌激素体外分析的方法，因为它具有良好的灵敏度，而且比其他分析方法更便宜，而且速度更快。(C)2004年，爱思唯尔有限公司出版。
In order to enhance the sensitivity and the speed of the yeast estrogen screen (YES)-assay, which has been established in many laboratories for the determination of estrogenic activity of compounds and environmental samples, the LYES-assay., a modified version of the YES-assay including a digestion step with the enzyme lyticase, was developed. With the LYES-assay the estrogenic activities of natural (17beta-estradiol E2 and estrone), synthetic (17alpha-ethinylestradiol EE2) and pharmaceutical estrogens (diethylstilbestrol DES) as well as xenoestrogens (4-nonylphenol NP and five parabens) were determined and compared with the results obtained by other in vitro-assays namely the conventional YES-assay, the E-Screen-assay (MCF-7 breast tumor cell proliferation) and a receptor binding-assay (RB) with human estrogen receptors hER-alpha and hER-beta. In the case of E2 the LYES-assay had a significantly lower limit of quantification (LOQ) than the conventional YES-assay and ever. two orders of magnitude lower than the RB-assay. Compared to the E-Screen-assay the LOQ of the LYES-assay was almost one order of magnitude higher. The time required to perform the LYES-assay was as little as seven hours compared to three to five days for the conventional YES-assay. Thus, the LYES-assay is a very good alternative to existing estrogenic in vitro-assays, since it has a good sensitivity, is cheap and much faster than the other assays. (C) 2004 Published by Elsevier Ltd.