A rapid reporter assay for recombinant human brain natriuretic peptide(rhBNP)by GloSensor technology

作     者：Lei Yu Xinchang Shi Chunmei Han Chunming Rao Junzhi Wang 

作者机构：National Institutes for Food and Drug ControlBeijing 100050China WHO CoUahoration Centre for Biologicals Standardization and EvaluationBeijing 100050China 

出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)

年 卷 期：2018年第8卷第5期

页      面：297-301页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 

基　　金：supported by grants from the National Science and Technology Major Project(No.2015ZX09501008-001) the Middle-aged and Young Development Research Foundation of NIFDC(No.2017B3) 

主　　题：RhBNP cGMP GloSensor technology Reporter assay 

摘      要：Accurate determination of biological activity is essential in quality control of recombinant human brain natriuretic peptide （rhBNP）. In previous study, we successfully developed a genetically modified cell line 293GCAC3-based ELISA assay for rhBNP. But ELISA procedure is still tedious, so this study was aimed to develop a rapid and simple bioassay for rhBNP using GloSensor technology, which provides a platform of flexible luciferase-based biosensors for real-time detection of signaling events in live cells, including cGMP production. A reporter cell line 293GCAGIo-G1 was constructed by transfecting pGloSensorTM 40 F plasmid into 293GCAC3. The reporter assay based on 293GCAGIo-G1 showed high precision with intraassay CV being 8.3% and inter-assay CV being 14.1%; high accuracy with 80%, 100% and 120% recovery rate being 99.2%, 102.4% and 99.0% respectively; and great linearity with R^2 of linear fitting equation being 0.99. Besides, no significant difference was found in test results of reporter assay and 293GCAC3-based ELISA assay （paired t test,p = 0.630）. All these results suggested that the reporter assay was a viable assay for biological determination of rhBNP.