Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline
Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline作者机构:Key Laboratory of Medical Laboratory Diagnostics of Ministry of Education Chongqing Medical University Chongqing 400016 China Unit for Biotransformation and Protem Blotechnology Chongqing Key Laboratory of Bioehemlcal and Molecular Pharmacology Chongqing Medical University Chongqing 400016 China
出 版 物:《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 (浙江大学学报(英文版)B辑(生物医学与生物技术))
年 卷 期:2011年第12卷第3期
页 面:180-188页
核心收录:
学科分类:1010[医学-医学技术(可授医学、理学学位)] 10[医学]
基 金:Project supported by the National Natural Science Foundation of China (No.30200266) the Program for New Century Excellent Talents in University of Ministry of Education of China(No.NCET-09-928)
主 题:Integration strategy Chromogenic substrate Data processing γ-Glutamyltransferase Kinetic analysis Serum enzyme assay
摘 要:At 0.12 mmol/L γ-glutamyl p-nitroaniline(GGPNA),an improved integrated method was developed for kinetic analysis of γ-glutamyltransferase(GGT) reaction process and the integration with the classical initial rate method to measure serum *** the improved integrated method,an integrated rate equation,which used the predictor variable of reaction time and considered inhibitions by both GGPNA and products,was nonlinearly fit to GGT reaction *** the integration strategy,classical initial rates were estimated when GGPNA consumption percentages were below 50%;otherwise,maximal reaction rates of GGT were estimated by the improved integrated method and converted into initial rates according to the differential rate equation at 0.11 mmol/L *** inte-gration strategy was validated using optimized GGT kinetic parameters and 10-s intervals to record reaction curves within 8.0 *** the integration strategy,there was a linear response from 0.9 to 32.0 U/L GGT,coefficients of variation were below 3.5%for GGT from 8.0 to 32.0 U/L(n=5) ,and GGT activities in clinical sera responded linearly to their classical initial rates at 2.00 mmol/L GGPNA with an expected ***,the integration strategy was successful in measuring GGT at 0.12 mmol/L GGPNA.