毛细管电泳
化学
色谱法
微量分析
蛋白质酪氨酸磷酸酶
磷酸酶
电泳
酶
生物化学
有机化学
作者
Xing‐Biao Qiu,Xuben Hou,Yue Yang,Hao Fang,Fei Cui,Yang Xia
标识
DOI:10.1016/j.chroma.2023.464511
摘要
Protein tyrosine phosphatase receptor type O (PTPRO) plays an important role in inflammation-related pathways and has become an emerging drug target. In this study, we developed an in-line capillary electrophoresis (CE) method for the investigation of the enzymatic activity of PTPRO, which was based on electrophoretically mediated microanalysis (EMMA). After a thorough method validation of the optimized conditions, this protocol was successfully employed to determine the kinetics of PTPRO as well as the half-maximal inhibitory concentration (IC50) of two typical PTPRO inhibitors. The final results were consistent with the values obtained through classical ultraviolet–visible (UV–vis) spectrophotometry. Our new method exhibited improved accuracy and reduced consumption, avoiding the disadvantages of traditional methods. This work provides a new strategy for PTPRO enzyme kinetic studies as well as inhibitory activity determination through capillary electrophoresis for the first time.
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