HPLC法测定rhNDPK-a酶活性  

Determination of recombinant human NDPK-a activity by HPLC

作　　者：钱垂文[1] 王一飞[1] 罗勇[1] 张美英[1] 熊盛[1] 

机构地区：[1]广州暨南大学药学院,广州510632

出　　处：《中国卫生检验杂志》2005年第4期390-391,共2页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金小额探索项目 (No 30371661);国家自然科学基金面上项目(No 30400071)

摘　　要：目的:研究不同反应体系对NDPK酶活性测定结果的影响。方法:分别以(UTP+ADP)和(ATP+UDP)作为反应底物,加入自制的rhNDPK-A蛋白,以100mmol/L磷酸二氢钾、20mmol/L乙酸、5mmol/L四丁基溴化铵,pH 5 0作为流动相A液, A液+25%乙腈作为B液,等梯度混合,C18柱,柱温25℃,流速1 0ml/min,检测波长254nm。结果:以(UTP+ADP)和(ATP+UDP)作为反应底物测定的酶比活分别为210U/mg、860U/mg。结论:不同反应体系对HPLC法测定NDPK酶活性有显著差异。Objective:To study the activity differences of NDPK in two reactive systems.Methods:HPLC-UV determination was employed with C_(18) column at 25℃. The mobile phase A was 100 mmol/L KH_2PO_4, 20 mmol/L actetic acid, 5 mmol/L tetrabutylamino bromide, pH 5.0. The mobile phase B was 25% acetonitrile dissolved in mobile phase A. The wave length for determination was 254 nm.Results:The specificactivity of rhNDPK-a in (UTP+ADP) and (ATP+UDP) systems was 210 U/mg and 860 U/mg respectively.Conclusion:There was striking difference between (UTP+ADP) and (ATP+UDP) systems in measuring NDPK activity by HPLC.

关 键 词：HPLC法 rhNDPK-a 酶活性 NDPK-A蛋白 mol/L 四丁基溴化铵 反应体系 磷酸二氢钾 测定结果 C18柱 检测波长 流动相 min 底物 A液 mg 

分 类 号：R286.0[医药卫生—中药学] R392.11[医药卫生—中医学]