Chemiluminescent assay of enzymes using proenhancers and pro-anti-enhancers.
Enhanced chemiluminescent assays for hydrolase enzymes have been developed using proenhancer and pro-anti-enhancer substrates. Alkaline phosphatase is measured using disodium para-iodophenyl phosphate (proenhancer) which is converted to para-iodophenol and this in turn enhances the light emission from the horseradish peroxidase catalysed chemiluminescent oxidation of luminol by peroxide. An alternative strategy uses para-nitrophenyl phosphate which is converted by alkaline phosphatase to para-nitrophenol which inhibits the enhanced chemiluminescent reaction. The detection limit for the enzyme using the proenhancer and pro-anti-enhancer assays was 100 attomoles and 1 picomole, respectively. The proenhancer strategy was effective in assays for beta-D-galactosidase, beta-D-glucosidase and aryl sulfatase. A limited comparison of the proenhancer and a conventional colorimetric assay for an alkaline phosphatase label in an enzyme immunoassay for alpha-fetoprotein showed good agreement.
Kricka
L J
LJ
Department of Pathology and Laboratory Medicine, Hospital of the University of Pennsylvania, Philadelphia 19104.
Schmerfeld-Pruss
D
D
Edwards
B
B
D000469:Alkaline Phosphatase / Q000032:analysis; D001192:Arylsulfatases / Q000032:analysis; D004798:Enzymes / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000187:drug effects* / Q000378:metabolism; D008163:Luminescent Measurements; D009596:Nitrophenols / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D013268:Stimulation, Chemical; D000509:alpha-Fetoproteins / Q000032:analysis; D001616:beta-Galactosidase / Q000032:analysis; D001617:beta-Glucosidase / Q000032:analysis
D004798:Enzymes; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D000509:alpha-Fetoproteins; C008644:nitrophenylphosphate; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase; D001192:Arylsulfatases; D001617:beta-Glucosidase; D001616:beta-Galactosidase