Abstract Preview Page

This is an automatically generated preview of the abstract you selected.

New bioluminescent assay of alkaline phosphatase and its application to EIA Hidetoshi Arakawa*; M Shiokawa; N Tajima; A Kokado; M Maeda Sch. of Pharm. Sci., Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo, 142-8555, JAPAN      *(arakawa@pharm.showa-u.ac.jp) We have developed a new bioluminescent assay of alkaline phosphatase (ALP) using ATP sulfurylase and luciferin luciferase reaction. Adenosine-3'-phosphate-5'-phosphosulfate (APSP) was used as substrate for ALP, which is hydrolyzed to produce APS. APS is converted to ATP by ATP sulfurylase in the presence of pyrophosphate. The produced ATP is determined by luciferin-luciferase reaction. The assay was performed as follows: 10 µl of ALP sample solution was mixed with 50 µl of APSP solution and incubated for 120 min. at 37 °C, and then 10 µl of pyrophosphate - ATP sulfurylase mixtures, and 100 µl of a luciferin-luciferase solution were added successively. The bioluminescent intensity was measured for 10-s after 15 min of addition of luciferin-luciferase solution. The standard curve of ALP was obtained in the range of 10 -21 mol to 10 -16 mol/assay, the detection limit was 10 -20 mol/assay. The coefficient of variation (CV) was examined at each point of the standard curve. The mean CV percentage was 4.47 % (n=6). This assay system was applied to enzyme immunoassay of hCG using ALP as label enzyme. The hCG in human urine could be assayed sensitively and directly by the proposed method. [Poster: arakawa.hideto.12122]

Return to the list of abstracts

E-mail   |  Conference Info  |   Index of Abstracts