3-磷酸甘油醛脱氢酶(GAPD)动力学方法的初步研究

目的 建立一种简便、快捷、准确的GAPD的检测方法,为以后的临床推广奠定基础.方法 以三乙醇胺作为缓冲液(PH为7.4),1,3-二磷酸甘油酸为底物,偶联3-磷酸甘油酸激酶,还原型辅酶Ⅰ(NADH)被氧化成氧化型辅酶Ⅰ (NAD+),在波长340nm处检测吸光度的变化值,在一定范围内,吸光度的下降与GAPD的活性成正...

Preliminary study of a kinetic method of Glyceraldehyde-3-phosphate dehydrogenase determination

【Objective】 To establish a simple rapid and accurate kinetic monitoring assay for detecting Glyceraldehyde-3-phosphate dehydrogenase（GAPD）.【Methods】 1,3-diphosphoglycerate was as substrate,coupling 3-phosphoglyceric phosphokinase,GAPD catalyzed the substrate in triethanolamine buffe,after a series of enzyme pair association response;NADH＋H＋ was oxidized to NAD＋.The activity of GAPD was determined at 340nm.【Results】 The most appropriate PH is 7.4 and Km is 6.3×10-4mmol/L;the optional substrate concentration was 0.007mmol/L.Inter-assay coefficient of variation（CV） value was 1.60% and intra-assay CV was 1.94%.The linearity reached 0-140U/L.The normal reference value（180 healthy subjects） was 4-28U/L.【Conclusion】 This method for GAPD determination is rapid,accurate and suitable for any kind of automatic biochemistry analyze.