Novel Phenothiazine Enhancers In Chemiluminescent Enzyme Immunoassay | 18699
Journal of Analytical & Bioanalytical Techniques
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Commonly in chemiluminescent enzyme immunoassay (CL-EIA) a light is formed upon the oxidation of luminol with
hydrogen peroxide catalyzed by peroxidase used as a label of immunoreagents. Since a peroxidase is poor catalyst in
this reaction, enhancers are added to increase CL intensity. For long time the most popular enhancer was 4-iodophenol. In
this work we analyzed some phenothiazine derivatives and showed that phenothiazines carrying groups with negative charge
are potent primary enhancers in peroxidase-catalyzed CL, whereas phenothiazines with positive charge have no enhancing
ability. The most efficient primary enhancers, whose the enhancing activity is higher many times than that of p-iodophenol,
are 3-(10�-phenothiazinyl)propane-1-sulfonate (SPTZ) and 3-(10�-phenothiazinyl)propionic acid (PPA). Some pyridine
derivatives are secondary enhancers increasing the enhancing ability of phenothiazines. Screening of some pyridines showed
that N-morpholinopyridine (MORPH) is the most active secondary enhancer. The mechanism of its enhancing action was
proposed. The conditions of the performance of the enhanced CL reaction using SPTZ or PPA in combination with MORPH
as primary and secondary enhancers were optimized. The detection systems with SPTZ/PPA and MORPH were applied
successfully in construction of ultrasensitive EIA kit for determination of human thyroglobulin and methylglyoxal-modified
low density lipoprotein. The obtained results open good perspectives for use of ECR with phenothiazines/MORPH in the
development of ultra-sensitive immunoassay kits.
Ivan Yu Sakharov is a Leading Scientific Researcher at the Department of Chemistry, Lomonosov Moscow State University (Russia). He has graduated with PhD degree in Kinetics and Catalysis in 1982 and DSc degree in Biotechnology in 1992. Presently his work focuses on the development of novel sensitive chemiluminescent methods for their use in analytical practice. He has published more than 147 papers in peer-reviewed journals.
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