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Development of a highly sensitive chemiluminescent assay for hydrogen peroxide under neutral conditions using acridinium ester and its application to an enzyme immunoassay
Author(s) -
Arakawa Hidetoshi,
Tsuruoka Keiko,
Ohno Kenichi,
Tajima Noriko,
Nagano Hiromi
Publication year - 2014
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/bio.2555
Subject(s) - chemiluminescence , hydrogen peroxide , chemistry , immunoassay , pmac , enzyme , chromatography , substrate (aquarium) , chemiluminescent immunoassay , biochemistry , antibody , medicine , biology , electrical engineering , immunology , control system , engineering , ecology
We developed a highly sensitive chemiluminescent (CL) assay for hydrogen peroxide using 10‐methyl‐9‐(phenoxycarbonyl) acridinium fluorosulfonate (PMAC) that produced chemiluminescence under neutral conditions and applied it to an enzyme immunoassay (EIA). One picomole of hydrogen peroxide could be detected using the optimized PMAC‐CL method and 6.2 × 10 ‐20 mol β‐ d ‐galactosidase (β‐gal) could be detected by combining an indoxyl derivative substrate and the proposed PMAC‐CL method. This highly sensitive CL β‐gal assay was applied to an EIA for thyroid‐stimulating hormone (TSH) using β‐gal as a label enzyme; 0.02–100.0 μU/mL TSH in human serum could be assayed directly and with high reproducibility. Copyright © 2013 John Wiley & Sons, Ltd.