文章基本信息

标题：Analysis of Phosphatase Activity in a Droplet-Based Microfluidic Chip
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作者：Bala Murali Krishna Vasamsetti ; Yeon-Jun Kim ; Jung Hoon Kang 等
期刊名称：Biosensors
电子版ISSN：2079-6374
出版年度：2022
卷号：12
期号：9
DOI：10.3390/bios12090740
语种：English
出版社：MDPI Publishing
摘要：We report analysis of phosphatase activity and inhibition on droplet-based microfluidic chips. Phosphatases are such attractive potential drug targets because abnormal phosphatase activity has been implicated in a variety of diseases including cancer, neurological disorders, diabetes, osteoporosis, and obesity. So far, several methods for assessing phosphatase activity have been reported. However, they require a large sample volume and additional chemical modifications such as fluorescent dye conjugation and nanomaterial conjugation, and are not cost-effective. In this study, we used an artificial phosphatase substrate 3- O-methylfluorescein phosphate as a fluorescent reporter and dual specificity phosphatase 22. Using these materials, the phosphatase assay was performed from approximately 340.4 picoliter (pL) droplets generated at a frequency of ~40 hertz (Hz) in a droplet-based microfluidic chip. To evaluate the suitability of droplet-based platform for screening phosphatase inhibitors, a dose–response inhibition study was performed with ethyl-3,4-dephostatin and the half-maximal inhibitory concentration (IC 50) was calculated as 5.79 ± 1.09 μM. The droplet-based results were compared to microplate-based experiments, which showed agreement. The droplet-based phosphatase assay proposed here is simple, reproducible, and generates enormous data sets within the limited sample and reagent volumes.
关键词：droplet-based microfluidics;microdroplet;phosphatase activity;PTP assay;DUSP22;ethyl-3,4-dephostatin