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Chemiluminescent measurement of peroxidase activity and its application using a lucigenin CT‐complex †
Author(s) -
Katsuragi Hisashi,
Takahashi Kiyoshi,
Suzuki Hideaki,
Maeda Masako
Publication year - 2000
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/(sici)1522-7243(200001/02)15:1<1::aid-bio561>3.0.co;2-7
Subject(s) - lucigenin , chemiluminescence , reagent , chemistry , luminol , chromatography , detection limit , peroxidase , horseradish peroxidase , enzyme , biochemistry , organic chemistry , superoxide
Luminol (3‐aminophthaloylhydrazine) is well known as a chemiluminescent reagent that can be used to measure peroxidase (POD) activity. We have developed a more sensitive assay for POD using a new chemiluminescent reagent based on 10,10′‐dimethyl‐9,9′‐biacridinium nitrate (lucigenin). This new reagent was obtained from lucigenin by light irradiation in an organic polar solvent such as N,N‐dimethylacetamide (DMAc). The detection limit for POD activity using this reagent is 10 −19 mol/assay. In an application for enzyme immunoassay based on a POD label, we were able to detect human α‐fetoprotein (AFP) at 1pg/mL. The characterization of this chemiluminescent reagent was obtained by optical measurements, and the formation of lucigenin charge transfer complex (CT‐complex) was confirmed. We also investigated the stability of lucigenin CT‐complex as a chemiluminescent reagent for the measurement of POD activity, and confirmed that the reagent was stable for more than a year. Copyright © 2000 John Wiley & Sons, Ltd.