Chemiluminescent assay of various enzyme activites and its application to enzyme immunoassays

✍ Scribed by Maeda, Masako ;Arakawa, Hidetoshi ;Tsuji, Akio

• Publisher: John Wiley and Sons
• Year: 1989
• Weight: 602 KB
• Volume: 4
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170040121

✦ Synopsis

A highly sensitive chemiluminescent assay for NAD(P)H have been developed. The principle of the method is as follows; NAD(P)H reduces molecular oxygen t o superoxide anion (0;) and hydrogen peroxide (H202) in the presence of I-methoxy-5-methylphenazinium methyl sulphate (I-MPMS) as electron mediator. The produced 0; and HzOz can be measured by chemiluminescent reaction using isoluminol (IL) and microperoxidase (m-POD). A linear relationship between chemiluminescence intensity and NAD(P)H concentration (log/log) was obtained ranged from lo-' mol/l t o lo-' mol/l. This chemiluminescent reaction has been coupled t o the assay of glucose-6-phosphate dehydrogenase (GGPDH), Po-galactosidase (/%Gal) and alkaline phosphatase (ALP). The detection limits of GGPDH, fJ-Gal and ALP were lo-'" mol, mot and lo-'" mot per assay, respectively. The chemiluminescent assay of these enzymes applied t o chemiluminescent enzyme immunoassay for 17a-hydroxyprogesterone and DNA hybridization assay using these enzymes as label.