基于微孔板化学发光法检测碱性磷酸酶的研究

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:125 μtmol/L的CDP- Star在pH 9.5缓冲液中,37℃下与ALP反应10min.在此条件下,△RLU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.

Research of Chemiluminescent Assay of Alkaline Phosphatase Based on Microplate

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.