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一种新型的生物发光纳米荧光素酶酵母 - 雌激素筛选生物传感器(nanoYES),配备紧凑型无线摄像头,用于基于效应的内分泌干扰化学物质检测。

A novel bioluminescent NanoLuc yeast-estrogen screen biosensor (nanoYES) with a compact wireless camera for effect-based detection of endocrine-disrupting chemicals.

作者信息

Cevenini Luca, Lopreside Antonia, Calabretta Maria Maddalena, D'Elia Marcello, Simoni Patrizia, Michelini Elisa, Roda Aldo

机构信息

Department of Chemistry "G. Ciamician", Alma Mater Studiorum, University of Bologna, via Selmi 2, 40126, Bologna, Italy.

Gabinetto Regionale di Polizia Scientifica per l'Emilia Romagna, Via Volto Santo 3, 40123, Bologna, Italy.

出版信息

Anal Bioanal Chem. 2018 Feb;410(4):1237-1246. doi: 10.1007/s00216-017-0661-7. Epub 2017 Sep 30.

DOI:10.1007/s00216-017-0661-7
PMID:28965124
Abstract

The presence of chemicals with estrogenic activity in surface, groundwater, and drinking water poses serious concerns for potential threats to human health and aquatic life. At present, no sensitive portable devices are available for the rapid monitoring of such contamination. Here, we propose a cell-based mobile platform that exploits a newly developed bioluminescent yeast-estrogen screen (nanoYES) and a low-cost compact camera as light detector. Saccharomyces cerevisiae cells were genetically engineered with a yeast codon-optimized variant of NanoLuc luciferase (yNLucP) under the regulation of human estrogen receptor α activation. Ready-to-use 3D-printed cartridges with immobilized cells were prepared by optimizing a new procedure that enables to produce alginate slices with good reproducibility. A portable device was obtained exploiting a compact camera and wireless connectivity enabling a rapid and quantitative evaluation (1-h incubation at room temperature) of total estrogenic activity in small sample volumes (50 μL) with a LOD of 0.08 nM for 17β-estradiol. The developed portable analytical platform was applied for the evaluation of water samples spiked with different chemicals known to have estrogen-like activity. Thanks to the high sensitivity of the newly developed yeast biosensor and the possibility to wireless connect the camera with any smartphone model, the developed configuration is more versatile than previously reported smartphone-based devices, and could find application for on-site analysis of endocrine disruptors. Graphical abstract Wireless effect-based detection of endocrine-disrupting chemicals with nanoYES platform.

摘要

地表、地下及饮用水中具有雌激素活性的化学物质的存在,对人类健康和水生生物构成了潜在威胁,引发了人们的严重担忧。目前,尚无灵敏的便携式设备可用于快速监测此类污染。在此,我们提出一种基于细胞的移动平台,该平台利用新开发的生物发光酵母 - 雌激素筛选方法(nanoYES)以及低成本的紧凑型相机作为光探测器。酿酒酵母细胞经基因工程改造,在人雌激素受体α激活的调控下表达酵母密码子优化的纳米荧光素酶变体(yNLucP)。通过优化一种新方法制备了固定有细胞的即用型3D打印盒,该方法能够生产具有良好重现性的藻酸盐切片。利用紧凑型相机和无线连接获得了一种便携式设备,可在室温下孵育1小时,对小体积样品(50 μL)中的总雌激素活性进行快速定量评估,对17β - 雌二醇的检测限为0.08 nM。所开发的便携式分析平台用于评估添加了已知具有雌激素样活性的不同化学物质的水样。由于新开发的酵母生物传感器具有高灵敏度,且相机可与任何智能手机型号进行无线连接,因此所开发的配置比先前报道的基于智能手机的设备更具通用性,可用于内分泌干扰物的现场分析。图形摘要:基于无线效应的nanoYES平台检测内分泌干扰化学物质。

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