基于微孔板化学发光法检测碱性磷酸酶的研究  

Research of Chemiluminescent Assay of Alkaline Phosphatase Based on Microplate

作　　者：王宇[1] 王宗良[1] 崔立国[1] 庄秀丽[1] 陈学思[1] 章培标[1] 

机构地区：[1]中国科学院长春应用化学研究所、生态环境高分子材料重点实验室,长春130021

出　　处：《分析化学》2012年第8期1279-1283,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.50733003;50973109);科技部国际合作((No.2010DFB50890)项目资助

摘　　要：CDP-Star能被ALP迅速水解,且水解产物能够持续发光。本研究对化学发光法检测ALP的反应条件进行了优化。优化后的反应条件为:125μmol/L的CDP-Star在pH 9.5缓冲液中,37℃下与ALP反应10min。在此条件下,ΔRLU与ALP的活力呈线性关系。因此,建立了一种基于微孔板化学发光法检测ALP的实验方法。结果表明,本方法检测ALP的检测线性范围是0.05～10 U/L和10～1000 U/L,相对于光吸收法具有速度快、灵敏度高、重复性好等优点。因此,本方法可以被开发成试剂盒,应用于高通量全自动生化分析仪的测试分析。Enzymatic dephosphorylation of CDP-Star by ALP leads to CDP-Star decompose and continuously emit light.Optimal conditions for the determination of ALP by chemiluminescent method were investigated in this research.125 μmol/L CDP-Star reacted with ALP in the buffer solution of pH=9.5 and incubated for 10 min at 37 ℃ were selected as the optimal conditions for the determination of ALP.The amount of ΔRLU is proportional to ALP activity.Therefore,the method of chemiluminescent assay of ALP based on microplate was established.The results showed that the linear range for the determination of ALP was 0.05-10 U/L and 10-1000 U/L.This method was more rapid,sensitive and reproducible than that of traditional colorimetric method.Therefore,this method can be developed into a kit used in the high-throughput automatic biochemical analyzer.

关 键 词：碱性磷酸酶 CDP-Star 化学发光 

分 类 号：O657.3[理学—分析化学]