Engineered Yeast Displaying Specific Norovirus-Binding Nanobodies for the Concentration and Detection of Human Norovirus in Food Matrix.
Engineered Yeast Displaying Specific Norovirus-Binding Nanobodies for the Concentration and Detection of Human Norovirus in Food Matrix.
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DOI：
10.1021/acs.jafc.3c01946
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发表时间：
2023-06-07
影响因子：
6.1
通讯作者：
Chen, Juhong
中科院分区：
文献类型：
--
作者：
Zhao, Xue;Rahman, Mahbubur;Xu, Zhiyuan;Kasputis, Tom;He, Yawen;Yuan, Lijuan;Wright, R. Clay;Chen, Juhong
人类诺如病毒对公共卫生和经济构成严重威胁。在这项研究中，我们对酵母（Saccharomyces cerevisiae EBY100）进行了基因工程改造，使其在细胞表面显示特异性的诺如病毒结合纳米体（Nano-26和Nano-85），以促进诺如病毒的浓度，从而提高检测效率。利用共聚焦显微镜和流式细胞术证实并表征了诺如病毒样颗粒（vlp）与这些纳米体显示酵母的结合。我们的工程酵母捕获诺如病毒VLPs的能力可达91.3%。此外，该方法还应用于诺瓦克病毒VLPs在真实食物基质中的浓缩和检测。检测范围为1 ~ 104 pg/g，检测限低至0.071 pg/g。总的来说，我们的工程酵母可能是一种很有前途的方法，可以浓缩和纯化食品样品中的诺如病毒，以便于检测，这使我们能够防止食源性病毒在食品供应链中的传播。
Human noroviruses pose grave threats to public health and economy. In this study, we genetically engineered yeast (Saccharomyces cerevisiae EBY100) to display specific norovirus-binding nanobodies (Nano-26 and Nano-85) on cell surface to facilitate the concentration of noroviruses for improved detection. Binding of norovirus virus-like particles (VLPs) to these nanobody-displaying yeasts was confirmed and characterized using confocal microscopy and flow cytometry. The ability of our engineered yeasts to capture norovirus VLPs can reach up to 91.3%. Furthermore, this approach was applied to concentrate and detect norovirus VLPs in a real food matrix. A wide linear detection range (1−104 pg/g) was observed, and the detection limit on spiked spinach was calculated as low as 0.071 pg/g. Overall, our engineered yeasts could be a promising approach to concentrate and purify noroviruses in food samples for easy detection, which allows us to prevent the spread of food-borne virus in the food supply chain.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
DOI：
10.1083/jcb.201409074
发表时间：
2015-06-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Helma J;Cardoso MC;Muyldermans S;Leonhardt H
通讯作者：
Leonhardt H
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
DOI：
10.1007/978-1-4939-2748-7_1
发表时间：
2015-01-01
期刊：
YEAST SURFACE DISPLAY: METHODS, PROTOCOLS, AND APPLICATIONS
影响因子：
--
作者：
Angelini, Alessandro;Chen, Tiffany F.;Wittrup, K. Dane
通讯作者：
Wittrup, K. Dane
影响因子：
4.7
作者：
Hillson, Nathan J.;Rosengarten, Rafael D.;Keasling, Jay D.
通讯作者：
Keasling, Jay D.