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.