Chemiluminescent assay of enzymes using proenhancers and pro-anti-enhancers

✍ Scribed by Kricka, Larry J. ;Schmerfeld-Pruss, Deborah ;Edwards, Brooks

• Publisher: John Wiley and Sons
• Year: 1991
• Weight: 639 KB
• Volume: 6
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170060404

✦ Synopsis

Enhanced chemiluminescent assays for hydrolase enzymes have been developed using proehancer and pro-anti-enhancer substrates. Alkaline phosphatase is measured using disodium para-iodophenyl phosphate (proenhancer) which is converted t o para-iodophenol and this i n 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 t o paranitrophenol 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 i n assays for beta-D-galactosidase, beta-o-glucosidase and aryl sulfatase. A limited comparison of the proenhancer and a conventional colorimetric assay for an alkaline phosphatase label i n an enzyme immunoassay for alpha-fetoprotein showed good agreement.