Quality-control method for the determination of biological activity of engineered calcineurin subunit B

作     者：Xinchang Shi Huan Yang Li Xu Xiang Li Zongwen Huang Yudong Han Qun Wei Chunming Rao 

作者机构：Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products National Institutes for Food and Drug Control Department of Biochemistry and Molecular Biology Beijing Normal University Gene Engineering and Biotechnology Beijing Key Laboratory Haikou Qili Pharmaceutical CO.LTD. 

基　　金：supported by the National Important Novel Medicine Research Project (2012ZX09304010, 2013ZX09102062) the National Natural Science Foundation of China (31270849) 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2016年第59卷第6期

页      码：584-588页

摘      要：The aim of this study was to establish a quality-control method for calcineurin subunit B(CNB) biological activity determinations. CNB enhances the p-nitrophenylphosphate(p NPP) dephosphorylating activity of calcineurin subunit A Δ316 mutant(CNAΔ316). A series of CNB concentrations were fitted to a four-parameter equation to calculate the corresponding p NPP maximum dephosphorylation rates. Values were calculated based on biological activity references using a parallel line method. The method was then validated for accuracy, precision, linearity, linear range, sensitivity, specificity, and robustness. The recovery results were greater than 98%. Intra-plate precision was 6.7%, with inter-plate precision of 10.8%. The coefficient of determination was greater than 0.98. The linear range was 0.05–50 μg m L?1, with sensitivity of 50 μg m L?1. Tested cytokines did not induce CNAΔ316 dephosphorylation of p NPP. The chosen CNAΔ316 concentration range did not affect activity determinations.

主 题 词：calcineurin subunit B biological activity method establishment method validation quality control 

学科分类：0710[理学-生物科学类] 071010[071010] 081704[081704] 07[理学] 08[工学] 0817[工学-轻工类] 

核心收录：

D　O　I：10.1007/s11427-016-5037-2

馆 藏 号：203413316...