Application of the cellular oxidation biosensor to Toxicity Identification Evaluations for high-throughput toxicity assessment of river water
Application of the cellular oxidation biosensor to Toxicity Identification Evaluations for high-throughput toxicity assessment of river water
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DOI：
10.1016/j.chemosphere.2020.125933
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发表时间：
2020-05-01
期刊：
影响因子：
8.8
通讯作者：
Mori, Izumi C.
中科院分区：
文献类型：
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作者：
Ooi, Lia;Okazaki, Keisuke;Mori, Izumi C.
毒性鉴定评价是对复杂环境水样中有毒物质进行分类鉴定的有效方法。然而，由于毒性生物测定的通量有限，其扩展到更大的样本量受到限制。在这里，我们报告了TIE第一阶段高通量方法的发展。这种新开发的方法是由基于荧光的细胞氧化（CO）生物传感器与roGFP 2表达的细菌细胞在96孔微孔板格式制造的辅助。用这种新方法对Langat河流域的四个河水样品进行了评价，结果表明，这四个样品的污染物组成可以分为两类。整个毒性试验由2338次试验组成，在12 h内用荧光酶标仪完成。同时，每次测定的样品体积减少到50 μ L，与常规生物测定相比，其小600至4700倍。这意味着CO生物传感器辅助TIE第一阶段的吞吐量现在是可行的，用于构建一个大规模的毒性监测系统，这将覆盖整个流域尺度。(C)2020年，任作家。爱思唯尔有限公司出版
Toxicity Identification Evaluation (TIE) is a useful method for the classification and identification of toxicants in a composite environment water sample. However, its extension to a larger sample size has been restrained owing to the limited throughput of toxicity bioassays. Here we reported the development of a high-throughput method of TIE Phase I. This newly developed method was assisted by the fluorescence-based cellular oxidation (CO) biosensor fabricated with roGFP2-expressing bacterial cells in 96-well microplate format. The assessment of four river water samples from Langat river basin by this new method demonstrated that the contaminant composition of the four samples can be classified into two distinct groups. The entire toxicity assay consisted of 2338 tests was completed within 12 h with a fluorescence microplate reader. Concurrently, the sample volume for each assay was reduced to 50 mu L, which is 600 to 4700 times lesser to compare with conventional bioassays. These imply that the throughput of the CO biosensor-assisted TIE Phase I is now feasible for constructing a large-scale toxicity monitoring system, which would cover a whole watershed scale. (C) 2020 The Authors. Published by Elsevier Ltd.